Updated on 2024/10/18

写真a

 
KANZAKI Hiroshi
 
Organization
Faculty of Environmental, Life, Natural Science and Technology Special-Appointment Professor
Position
Special-Appointment Professor
Contact information
メールアドレス
External link

Degree

  • PhD (Agriculture) ( 1986.11   Kyoto University )

Research Interests

  • Enzyme Chemistry

  • 応用微生物学

  • Applied Microbiology

  • Natural Products Chemistry

  • 天然物化学

  • 酵素化学

Research Areas

  • Nanotechnology/Materials / Chemistry and chemical methodology of biomolecules

  • Life Science / Bioorganic chemistry  / 天然物有機化学 生物活性

  • Life Science / Applied biochemistry

  • Life Science / Applied microbiology  / 発酵 醸造 微生物変換

Education

  • Kyoto University    

    - 1986

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  • Kyoto University   農学研究科   農芸化学

    - 1986

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    Country: Japan

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  • Kyoto University    

    - 1981

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  • Kyoto University   農学部   農芸化学科

    - 1981

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    Country: Japan

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Research History

  • Okayama University   学術研究院環境生命自然科学学域   Professor

    2023.4

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  • Okayama University   学術研究院環境生命科学学域   Professor

    2022.4 - 2023.3

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  • - Professor,Graduate School of Environmental and life Science,Okayama University

    2018.4 - 2022.3

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  • - 岡山大学環境生命科学研究科 教授

    2018.4 - 2022.3

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  • Executive Director and Vice President,Okayama University,Okayama University

    2017 - 2018

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  • Okayama University

    2017 - 2018

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  • 岡山大学環境生命科学研究科 教授

    2012 - 2017

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  • Professor,Graduate School of Environmental and life Science,Okayama University

    2012 - 2017

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  • アメリカ公衆衛生研究所 博士研究員

    1988 - 1989

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  • Postdoctoral Fellowships of Japan Society for the Promotion of Science

    1986 - 1987

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  • Kyoto University   Faculty of Agriculture

    1986 - 1987

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Professional Memberships

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Committee Memberships

  • Japan Society for Bioscience, Biotechnology, and Agrochemistryociety   auditor  

    2023   

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    Committee type:Academic society

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  • Japan Society for Bioscience, Biotechnology, and Agrochemistry   Vice President  

    2021 - 2023   

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    Committee type:Academic society

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  • Japan Society for Bioscience, Biotechnology, and Agrochemistry   auditor  

    2019 - 2021   

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    Committee type:Academic society

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  • Japan Society for Bioscience, Biotechnology, and Agrochemistry   Director (Public Relations)  

    2013 - 2017   

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    Committee type:Academic society

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  • おかやまバイオアクティブ研究会   会長  

    2012   

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    Committee type:Academic society

    おかやまバイオアクティブ研究会

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  • NPO Chu-Shikoku Agritech   President  

    2009   

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    Committee type:Other

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Papers

  • 微生物変換で天然に存在しない生物活性物質を創生する Invited

    神崎 浩

    農薬環境科学研究   30   7 - 14   2023.9

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    Authorship:Lead author, Last author, Corresponding author   Language:Japanese   Publishing type:Research paper (conference, symposium, etc.)  

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  • 麴菌固体培養の特性を駆使する植物資源素材からの高機能化素材開発 Invited

    仁戸田照彦, 深野夏暉, 神崎 浩

    アグリバイオ   92 ( 8 )   744 - 748   2023.8

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  • Potential of lactic acid bacteria to produce functional fermented whey beverage with putative health promoting attributes Reviewed

    Jirayu Jitpakdee, Duangporn Kantachote, Hiroshi Kanzaki, Teruhiko Nitoda

    LWT   160   113269 - 113269   2022.4

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    Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.lwt.2022.113269

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  • Aerobic bioconversion of C-glycoside mangiferin into its aglycone norathyriol by an isolated mouse intestinal bacterium. Reviewed International journal

    Uswatun Hasanah, Kasumi Miki, Teruhiko Nitoda, Hiroshi Kanzaki

    Bioscience, biotechnology, and biochemistry   85 ( 4 )   989 - 997   2021.3

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Norathyriol is an aglycone of a xanthonoid C-glycoside mangiferin that possesses different bioactive properties useful for humans compared to mangiferin. Mangiferin is more readily available in nature than norathyriol; thus, efficient mangiferin conversion into norathyriol is desirable. There are a few reports regarding mangiferin C-deglycosylation because of the C-C bond resistance toward acid, alkaline, and enzyme hydrolysis. In this study, we isolated a mangiferin-deglycosylating bacterium strain KM7-1 from the mouse intestine. 16S rDNA sequencing indicated that KM7-1 belongs to the Bacillus genus. Compared to the taxonomically similar bacteria, the growth characteristic of facultative anaerobic and thermophilic resembled, yet only Bacillus sp. KM7-1 was able to convert mangiferin into norathyriol. Resting cells of Bacillus sp. KM7-1 obtained from aerobic cultivation at 50 °C showed high norathyriol formation from 1 m m of mangiferin. Norathyriol formation can be conducted either under aerobic or anaerobic conditions, and the reaction depended on time and bacterial amount.

    DOI: 10.1093/bbb/zbaa121

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  • New analogs of pochonicine, a potent β-N-acetylglucosaminidase inhibitor from fungus Pochonia suchlasporia var. suchlasporia TAMA 87. Reviewed

    Yuhichi Mushiake, Aya Tsuchida, Asami Yamada, Hiroshi Kanzaki, Toru Okuda, Teruhiko Nitoda

    Journal of pesticide science   46 ( 1 )   115 - 119   2021.2

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    Three novel analogs of pochonicine (1) were isolated from a solid fermentation culture of the fungal strain Pochonia suchlasporia var. suchlasporia TAMA 87, and their structures were elucidated as 7-deoxypochonicine (2), 6-deoxypochonicine (3), and 6,7-dideoxypochonicine (4). These analogs were found to possess the same stereochemistry as pochonicine. Comparison of β-N-acetylglucosaminidase (GlcNAcase) inhibitory activity between these analogs and pochonicine suggested that the C-6 hydroxy group of pochonicine was essential to its potent GlcNAcase inhibitory activity and that the C-7 hydroxy group also contributed to the activity, but to a lesser extent than the C-6 hydroxy group.

    DOI: 10.1584/jpestics.D20-086

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  • Selected probiotic lactic acid bacteria isolated from fermented foods for functional milk production: Lower cholesterol with more beneficial compounds Reviewed

    Jirayu Jitpakdee, Duangporn Kantachote, Hiroshi Kanzaki, Teruhiko Nitoda

    LWT - Food Science and Technology   135   110061 - 110061   2021.1

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    Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.lwt.2020.110061

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  • Adipogenesis Suppression by Enokitake Mushroom Bed Extract and its Application to Pet Animal Supplements

    Bio Industry   38 ( 2 )   2021

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    J-GLOBAL

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  • A new asteltoxin analog with insecticidal activity from Pochonia suchlasporia TAMA 87. Reviewed

    Syaefudin Suminto, Eri Takatsuji, Ayako Iguchi, Hiroshi Kanzaki, Toru Okuda, Teruhiko Nitoda

    Journal of pesticide science   45 ( 2 )   81 - 85   2020.5

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    A new asteltoxin analog, named asteltoxin H (1), was isolated by the solid-state fermentation of the fungus Pochonia suchlasporia var. suchlasporia TAMA 87. The chemical structure of 1 was deduced by spectroscopic methods, including 1D and 2D NMR, HRESIMS, and UV-Vis analyses. Compound 1 showed insecticidal activity against prepupae of the blowfly, Lucilia sericata, with an LD50 value of 0.94 µg/mg prepupal body weight.

    DOI: 10.1584/jpestics.D19-081

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  • Efficient production of recombinant tannase in Aspergillus oryzae using an improved glucoamylase gene promoter. Reviewed

    Kyotaro Ichikawa, Yoshihito Shiono, Tomoko Shintani, Akira Watanabe, Hiroshi Kanzaki, Katsuya Gomi, Takuya Koseki

    Journal of bioscience and bioengineering   129 ( 2 )   150 - 154   2020.2

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    A tannase-encoding gene, AotanB, from Aspergillus oryzae RIB40 was overexpressed in A. oryzae AOK11 niaD-deficient mutant derived from an industrial strain under the control of an improved glucoamylase gene promoter PglaA142. The recombinant tannase, designated as rAoTanBO, was produced efficiently as an active extracellular enzyme. Purified rAoTanBO showed a smeared band with a molecular mass of approximately 80-100 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The rAoTanBO had a molecular mass of 65 kDa, after treatment with endo-β-N-acetylglucosaminidase H. Purified rAoTanBO exhibited maximum activity at 30-35°C and pH 6.0. The tannase activity of purified rAoTanBO towards natural and artificial substrates was 2-8 folds higher than that of the recombinant enzyme produced by Pichia pastoris, designated as rAoTanBP. N-terminus of the mature rAoTanBP had six more amino acids than the N-terminus of the mature rAoTanBO. Kinetic analyses showed that rAoTanBO had higher catalytic efficiency (kcat/Km) than rAoTanBP. rAoTanBO was stable up to 60°C and higher thermostability than rAoTanBP. N-linked oligosaccharides had no effect on the activity and stability of rAoTanBO and rAoTanBP.

    DOI: 10.1016/j.jbiosc.2019.08.002

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  • Optimization of culture conditions for production of antivibrio compounds from probiotic purple nonsulfur bacteria against acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus and Vibrio spp. Reviewed

    Supaporn Chumpol, Duangporn Kantachote, Pattamarat Rattanachuay, Salwa Torpee, Teruhiko Nitoda, Hiroshi Kanzaki

    Aquaculture   505   72 - 83   2019.4

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    DOI: 10.1016/j.aquaculture.2019.02.040

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  • Biotransformation of β-Mangostin by an Endophytic Fungus of Garcinia mangostana to Furnish Xanthenes with an Unprecedented Heterocyclic Skeleton. Reviewed International journal

    Panarat Arunrattiyakorn, Mayuso Kuno, Thammarat Aree, Surat Laphookhieo, Teerayut Sriyatep, Hiroshi Kanzaki, Miguel Angel Garcia Chavez, Yan Alexander Wang, Raymond J Andersen

    Journal of natural products   81 ( 10 )   2244 - 2250   2018.10

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    Biotransformation of β-mangostin (1) by the endophytic fungus Xylaria feejeensis GM06 afforded hexacyclic ring-fused xanthenes with an unprecedented hexacyclic heterocylic skeleton. β-Mangostin (1) was transformed to two diastereomeric pairs of enantiomers, mangostafeejin A [(-)-2a/(+)-2b)] and mangostafeejin B [(-)-3a/(+)-3b)]. The chemical structures of the transformation products were elucidated by analysis of NMR and MS data, and the structure of mangostafeejin A [(-)-2a/(+)-2b)] was confirmed by single-crystal X-ray diffraction analysis. The absolute configurations of 3a and 3b were established on the basis of calculated and measured ECD data using the ECD spectra of 2a and 2b as models. The fungal biotransformation described herein provides an effective method to convert an abundant achiral plant natural product scaffold into new chiral heterocyclic scaffolds representing expanded chemical diversity for biological activity screening.

    DOI: 10.1021/acs.jnatprod.8b00519

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  • Use of wood vinegar to enhance 5-aminolevulinic acid production by selected Rhodopseudomonas palustris in rubber sheet wastewater for agricultural use Reviewed

    Tomorn Nunkaew, Duangporn Kantachote, Sumate Chaiprapat, Teruhiko Nitoda, Hiroshi Kanzaki

    Saudi Journal of Biological Sciences   25 ( 4 )   642 - 650   2018.5

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    Authorship:Last author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier B.V.  

    This study aimed to produce inexpensive 5-aminolevulinic acid (ALA) in a non-sterile latex rubber sheet wastewater (RSW) by Rhodopseudomonas palustris TN114 and PP803 for the possibility to use in agricultural purposes by investigating the optimum conditions, and applying of wood vinegar (WV) as an economical source of levulinic acid to enhance ALA content. The Box–Behnken Design experiment was conducted under microaerobic-light conditions for 96 h with TN114, PP803 and their mixed culture (1:1) by varying initial pH, inoculum size (% v/v) and initial chemical oxygen demand (COD, mg/L). Results showed that the optimal condition (pH, % inoculum size, COD) of each set to produce extracellular ALA was found at 7.50, 6.00, 2000 for TN114
    7.50, 7.00, 3000 for PP803
    and 7.50, 6.00, 4000 for a mixed culture
    and each set achieved COD reduction as high as 63%, 71% and 75%, respectively. Addition of the optimal concentration of WV at mid log phase at 0.63% for TN114, and 1.25% for PP803 and the mixed culture significantly increased the ALA content by 3.7–4.2 times (128, 90 and 131 μM, respectively) compared to their controls. ALA production cost could be reduced approximately 31 times with WV on the basis of the amount of levulinic acid used. Effluent containing ALA for using in agriculture could be achieved by treating the RSW with the selected ALA producer R. palustris strains under the optimized condition with a little WV additive.

    DOI: 10.1016/j.sjbs.2016.01.028

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  • Administration of purple nonsulfur bacteria as single cell protein by mixing with shrimp feed to enhance growth, immune response and survival in white shrimp (Litopenaeus vannamei) cultivation Reviewed

    Supaporn Chumpol, Duangporn Kantachote, Teruhiko Nitoda, Hiroshi Kanzaki

    Aquaculture   489   85 - 95   2018.3

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    Single cell protein (SCP) is an alternative way to increase nutrients for animal consumption
    and purple nonsulfur bacteria (PNSB) should be considered as SCP due to their rich sources of protein, vitamins and photopigments. Hence, the aim of this study was to investigate the potential of promising PNSB to be used as SCP by mixing with commercial shrimp feed for white shrimp cultivation starting from postlarval until early juvenile stages for 60 days. PNSB strains, Rhodobacter sphaeroides SS15 and Afifella marina STW181 were selected, based on high amounts of biologically complete protein and photopigments, to use as SCP at a ratio of 1:1
    and their lyophilized cells at 1, 3 and 5% (w/w) were mixed well with commercial shrimp feed to obtain modified shrimp feed recipes
    Diet 1, Diet 2 and Diet 3, respectively. Levels of NH4 +, NO2 −, NO3 − and COD in rearing water from Diet 2 and 3 sets were significantly higher than control set. However, Diet 1 set showed the lowest levels of these water parameters among modified diet sets as no significant difference levels of NO3 − and COD between Diet 1 and control sets. Shrimp growth performance on the basis of relative gain rate and other growth parameters found that Diet 1 set was much better than sets of Diet 2, Diet 3 and control with the lowest found in the control set. In addition, the maximum shrimp survival was observed in Diet 1 set (85%) although no significant difference among them as 80% in control set. No significant difference was found among control and all modified shrimp diets for total hemocyte count in shrimp
    however, significant increases of superoxide dismutase activity found in sets of Diet 1, while phenoloxidase activity found in Diet 2 and Diet 3. The results of hepatopancreas (HP) histopathology analysis also showed a good condition of HP as healthy shrimp. PNSB biomass as SCP at optimal level has the potential to be an effective source of a novel protein in shrimp feed to enhance shrimp growth and also to increase shrimp survival as more nutritious with no effect on water quality.

    DOI: 10.1016/j.aquaculture.2018.02.009

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  • Unique antimicrobial spectrum of ophiobolin K produced by Aspergillus ustus Reviewed

    Natthapat Sohsomboon, Hiroshi Kanzaki, Teruhiko Nitoda

    Bioscience, Biotechnology and Biochemistry   82 ( 3 )   422 - 424   2018

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japan Society for Bioscience Biotechnology and Agrochemistry  

    A co-cultivation study of two fungal strains showed that Aspergillus ustus could inhibit Aspergillus repens growth. The bioactive compound responsible for the observed activity was purified and identified as a sesterterpene, ophiobolin K. Ophiobolin K exhibited marked inhibition against both fungi and bacteria, especially A. repens, A. glaucus and gram-positive bacteria including Bacillus subtilis, Staphylococcus aureus, and Micrococcus luteus.

    DOI: 10.1080/09168451.2018.1429890

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  • Development of Energy-saving Ventilation System Considering Inside and Outside Temperatures of a Sawdust-based Shiitake Cultivation Facility Reviewed

    柏野泰章, 明貝文夫, 難波和彦, 門田充司, 神崎浩

    美味技術学会誌   16 ( 2 )   2018

  • In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus and other vibrios Reviewed

    Supaporn Chumpol, Duangporn Kantachote, Pattamarat Rattanachuay, Varaporn Vuddhakul, Teruhiko Nitoda, Hiroshi Kanzaki

    AQUACULTURE RESEARCH   48 ( 6 )   3182 - 3197   2017.6

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    Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticus AHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticus SR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.

    DOI: 10.1111/are.13149

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  • The roles of probiotic purple nonsulfur bacteria to control water quality and prevent acute hepatopancreatic necrosis disease (AHPND) for enhancement growth with higher survival in white shrimp (Litopenaeus vannamei) during cultivation Reviewed

    Supaporn Chumpol, Duangporn Kantachote, Teruhiko Nitoda, Hiroshi Kanzaki

    AQUACULTURE   473   327 - 336   2017.4

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    This study aimed to investigate the potential of a mixed probiotic purple nonsulfur bacteria (PNSB) for controlling water quality and preventing acute hepatopancreatic necrosis disease (AHPND) to promote growth performance and increase survival of white shrimp (Litopenaeus vannamei) in postlarval stage during cultivation. Four probiotic PNSB (Rhodobacter sphaeroides strains SS15, S3W10, TKW17 and Afifella marina STW181) were investigated for controlling water quality using a mixed culture at a ratio of 1: 1 (roughly 1 x 10(8) cells mL(-1) for each, every week 1-7) as follows; T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) and found that throughout 8 weeks shrimp cultivation the most effective sets to reduce NH4(+) and promote shrimp growth (based on weight and total length) were T1 and T3, respectively. Hence, PNSB strains (SS15, S3W10 and STW181) were used as a mixed culture (1: 1: 1, roughly 1 x 10(8) cells mL(-1) for each at weeks 1, 2 and 3) assessing their potential to prevent AHPND by challenge test on shrimp with a virulent strain AHPND-causing Vibrio parahaemolyticus SR2 (roughly 1 x 10(5) cells mL(-1) at day 15) during shrimp cultivation for 30 days. Inoculated PNSB sets (positive control: only PNSB inoculation, and treatment: both PNSB and SR2 inoculations) significantly decreased (P < 0.05) the levels of NH4-, NO2-, NO3- and chemical oxygen demand (COD) with significantly higher (P < 0.05) of dissolved oxygen (DO) compared to native control (no inoculation) and challenge test (only SR2 inoculation). The maximal growth performance of shrimp was observed in the positive control; while no significant difference (P > 0.05) was observed for other sets. PNSB survived and colonized in intestinal shrimp tract to prevent AHPND by increasing 11% survival rate of infected shrimp by strain SR2. The correlation coefficient between PNSB population and shrimp survival showed a positive strong correlation, but a negative strong correlation between vibrios population and shrimp survival. Overall results proved that a mixed three probiotic PNSB are a good candidate for applying in white shrimp cultivation to maintain water quality and to protect shrimp diseases for promoting shrimp growth with higher survival rate. (C) 2017 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.aquaculture.2017.02.033

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  • Selection of salt tolerant purple nonsulfur bacteria producing 5-aminolevulinic acid (ALA) and reducing methane emissions from microbial rice straw degradation Reviewed

    Tomorn Nunkaew, Duangporn Kantachote, Teruhiko Nitoda, Hiroshi Kanzaki

    APPLIED SOIL ECOLOGY   86   113 - 120   2015.2

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    Saline soil is one major problem that causes low productivity on rice, while paddy fields are also one source of methane (CH4) emissions that can have a considerable impact on global warming. By using purple nonsulfur bacteria (PNSB) it is possible to overcome both these serious problems because PNSB produce 5-aminolevulinic acid (ALA), a compound that reduces salt stress for plants, and they are also good competitors for methanogens in the paddy fields. Therefore, the objective of this study was to select salt resistant PNSB with an ability to release ALA and reduce CH4 emissions. The PNSB isolates used were obtained from saline paddy fields in southern Thailand and the best were selected on the basis of their good growth under conditions of aerobic dark and anaerobic light while being salt tolerant, released ALA and reduced CH4 emissions under microaerobic light conditions. Among the 272 PNSB isolates, TN114 was the best ALA producer while PP803 was the best to reduce CH4 emissions. Both selected PNSB strains grew well in glutamate acetate (GA) broth containing NaCl up to 6.00% but 0.25% NaCl was their optimal concentration for growth. Under salt stress with 0.25% NaCl, the strain TN114 released 25.63 mu M ALA in GA broth. In scaled-up experiments, when it was mixed with native soil flora in rice straw broth the strain TN114 released 13.44 mu M ALA and the strain PP803 reduced CH4 emissions by 88.41%. These two strains were identified as Rhodopseudomonas palustris according to both traditional and 16 S rRNA identifications. Overall the results demonstrated that they had a good potential as plant growth stimulating bacteria under salt stress, due to the release of ALA and also reduce CH4 emissions. (C) 2014 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.apsoil.2014.10.005

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  • Characterization of exopolymeric substances from selected Rhodopseudomonas palustris strains and their ability to adsorb sodium ions Reviewed

    Tomorn Nunkaew, Duangporn Kantachote, Teruhiko Nitoda, Hiroshi Kanzaki, Raymond J. Ritchie

    CARBOHYDRATE POLYMERS   115 ( 22 )   334 - 341   2015.1

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    Removal of Na+ by binding with exopolymeric substances (EPS) from Rhodopseudomonas palustris TN114 and PP803 was investigated. The moderate negative correlation pairs (r(p)) between remaining Alcian blue and amount of Na+ adsorbed on EPS from strains TN114 and PP803 were -0.652 and -0.609. Both strains showed positive relationships between the amounts of EPS produced and bacterial growth. EPS from strain PP803 had a higher efficiency in removing Na+ than the EPS from strain TN114 based on their EC50 values (1.79 and 1.49 mg/mL for TN114 and PP803, respectively). The principal component from EPS of strain PP803 which was responsible for salt removal was purified and it was identified as a polysaccharide kDa) mainly composed of galacturonic acid. Overall results suggested that EPS is a key factor that our strains used to bind Na* allowing their survival in high NaC1 concentrations. (C) 2014 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.carbpol.2014.08.099

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  • Biotransformation of alpha-mangostin by Colletotrichum sp MT02 and Phomopsis euphorbiae K12 Reviewed

    Panarat Arunrattiyakorn, Nuttika Suwannasai, Thammarat Aree, Somdej Kanokmedhakul, Hideyuki Ito, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   102   174 - 179   2014.4

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    The microbial transformation of the major mangosteen pericarp xanthone, alpha-mangostin (1) by the endophytic fungi Colletotrichum sp. MT02 and Phomopsis euphorbiae K12 resulted in the production of the five metabolites 2-6. Biotransformation with Colletotrichum sp. MT02 converted 1 into 17,18-dihydroxymangostanin (2) and cyclomangostanin (3), while incubation with P. euphorbiae K12 gave 12,13,20-trihydroxymangostin (4), 12,13,19-trihydroxymangostin (5), and 20-hydroxymangostanin (6) as transformation products. The structures of these metabolites were elucidated via spectroscopic analyses, and for compound 3, the structure was confirmed by X-ray crystallographic analysis. All metabolites are new, and metabolite 3 possessed an unusual structure of bis-ring fused xanthene in nature. In addition, substrate and all products were evaluated for the antimycobacterial activity against Mycobacterium tuberculosis as well as the cytotoxicity against breast cancer (MCF-7) cell lines. (C) 2014 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.molcatb.2014.02.010

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  • Effects of 5-aminolevulinic acid (ALA)-containing supernatants from selected Rhodopseudomonas palustris strains on rice growth under NaCl stress, with mediating effects on chlorophyll, photosynthetic electron transport and antioxidative enzymes Reviewed

    Tomorn Nunkaew, Duangporn Kantachote, Hiroshi Kanzaki, Teruhiko Nitoda, Raymond J. Ritchie

    ELECTRONIC JOURNAL OF BIOTECHNOLOGY   17 ( 1 )   19 - 26   2014.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:UNIV CATOLICA DE VALPARAISO  

    Background: Rice is globally one of the most important food crops, and NaCl stress is a key factor reducing rice yield. Amelioration of NaCl stress was assessed by determining the growth of rice seedlings treated with culture supernatants containing 5-aminolevulinic acid (ALA) secreted by strains of Rhodopseudomonas palustris (TN114 and PP803) and compared to the effects of synthetic ALA (positive control) and no ALA content (negative control).
    Results: The relative root growth of rice seedlings was determined under NaCl stress (50 mM NaCl), after 21 d of pretreatment. Pretreatments with 1 mu M commercial ALA and 10X diluted culture supernatant of strain TN114 (2.57 mu M ALA) gave significantly better growth than 10X diluted PP803 supernatant (2.11 mu M ALA). Rice growth measured by dry weight under NaCl stress ordered the pretreatments as: commercial ALA N TN114 N PP803 N negative control. NaCl stress strongly decreased total chlorophyll of the plants that correlated with non-photochemical quenching of fluorescence (NPQ). The salt stress also strongly increased hydrogen peroxide (H2O2) concentration in NaCl-stressed plants. The pretreatments were ordered by reduction in H2O2 content underNaCl stress as: commercial ALA N TN114 N PP803 N negative control. The ALA pretreatments incurred remarkable increases of total chlorophyll and antioxidative activities of catalase (CAT), ascorbate peroxide (APx), glutathione reductase (GR) and superoxide dismutase (SOD); under NaCl stress commercial ALA and TN114 had generally stronger effects than PP803.
    Conclusions: The strain TN114 has potential as a plant growth stimulating bacterium that might enhance rice growth in saline paddy fields at a lower cost than commercial ALA. (C) 2014 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.

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  • Total synthesis of the proposed structure for pochonicine and determination of its absolute configuration Reviewed

    Yujiro Kitamura, Hiroyuki Koshino, Takemichi Nakamura, Aya Tsuchida, Teruhiko Nitoda, Hiroshi Kanzaki, Koji Matsuoka, Shunya Takahashi

    Tetrahedron Letters   54 ( 11 )   1456 - 1459   2013.3

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    Pochonicine is a polyhydroxylated pyrrolizidine alkaloid with a powerful inhibitory activity against β-Nacetylglucosaminidases. The proposed structure for pochonicine and the three diastereomers concerning its C-1 and/or C-3 positions were synthesized from N-acetyl-D-glucosamine through construction of the pyrrolizidine skeleton by two intramolecular amino cyclizations as key steps. This synthetic study not only revised the structure of the natural product to the corresponding 1,3-di-epi-form but also determined the absolute configuration as 1R, 3S, 5R, 6R, 7S, 7aR. Copyright © 2012 Published by Elsevier Ltd.

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  • TMG-chitotriomycin as a probe for the prediction of substrate specificity of β-N-acetylhexosaminidases Reviewed

    Hiroto Shiota, Hiroshi Kanzaki, Tadashi Hatanaka, Teruhiko Nitoda

    Carbohydrate Research   375   29 - 34   2013

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    TMG-chitotriomycin (1) produced by the actinomycete Streptomyces annulatus NBRC13369 was examined as a probe for the prediction of substrate specificity of β-N-acetylhexosaminidases (HexNAcases). According to the results of inhibition assays, 14 GH20 HexNAcases from various organisms were divided into 1-sensitive and 1-insensitive enzymes. Three representatives of each group were investigated for their substrate specificity. The 1-sensitive HexNAcases hydrolyzed N-acetylchitooligosaccharides but not N-glycan-type oligosaccharides, whereas the 1-insensitive enzymes hydrolyzed N-glycan-type oligosaccharides but not N-acetylchitooligosaccharides, indicating that TMG-chitotriomycin can be used as a molecular probe to distinguish between chitin-degrading HexNAcases and glycoconjugate-processing HexNAcases. © 2013 Elsevier Ltd. All rights reserved.

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  • The use of rice straw broth as an appropriate medium to isolate purple nonsulfur bacteria from paddy fields Reviewed

    Tomorn Nunkaew, Duangporn Kantachote, Teruhiko Nitoda, Hiroshi Kanzaki

    ELECTRONIC JOURNAL OF BIOTECHNOLOGY   15 ( 6 )   2012.11

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    The aims were to explore an appropriate isolating medium for obtaining purple nonsulfur bacteria (PNSB) for use as biofertilizers in saline paddy fields and to obtain pure cultures. We therefore chose a defined isolating medium containing 0.25% NaCl, (Glutamate-Acetate broth, GA) and a rice straw broth to compare them for numbers of PNSB obtained, time to obtain pure cultures, diversity and costs. A total of 30 water and 30 sediment samples were collected from saline paddy fields in southern Thailand and used to isolate PNSB in both the isolating media. Based on 60 samples and a period of 13 days incubation under anaerobic light conditions, a greater number of samples produced PNSB growth in GA broth after only day 3; however, after that the rice straw broth provided about a 2 fold increase in the number of samples that produced PNSB growth. Colonies isolated from GA broth required a significantly higher number of repeated streaking to obtain a pure culture (average 3.5) than those from rice straw broth (average 2.7) and the latter medium also produced significantly (P < 0.05) more isolates per sample. Sixty samples of water and sediment, from rice paddies with salinity (average, 3.43 +/- 0.67 mS/cm) and slight acidity (average, pH 5.84 +/- 0.42) provided 62 PNSB isolates by GA broth and 210 isolates by rice straw broth, and rice straw broth also produced a greater prevalence of PNSB. Estimates of the costs based on current prices of media, Gas Pak and electricity to obtain PNSB with the use of GA broth was roughly 6 times higher than for the rice straw broth.

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  • Microbial metabolism of alpha-mangostin isolated from Garcinia mangostana L. Reviewed

    Panarat Arunrattiyakorn, Sunit Suksamrarn, Nuttika Suwannasai, Hiroshi Kanzaki

    PHYTOCHEMISTRY   72 ( 8 )   730 - 734   2011.6

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    alpha-Mangostin (1), a prenylated xanthone isolated from the fruit hull of Garcinia mangostana L., was individually metabolized by two fungi, Colletotrichum gloeosporioides (EYL131) and Neosartorya spathulata (EYR042), repectively. Incubation of 1 with C. gloeosporioides (EYL131) gave four metabolites which were identified as mangostin 3-sulfate (2), mangostanin 6-sulfate (3), 17,18-dihydroxymangostanin 6-sulfate (4)and isomangostanin 3-sulfate (5). Compound 2 was also formed by incubation with N. spathulata (EYR042). The structures of the isolated compounds were elucidated by spectroscopic data analysis. Of the isolated metabolites, 2 exhibited significant anti-mycobacterial activity against Mycobacterium tuberculosis. (C) 2011 Elsevier Ltd. All rights reserved.

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  • Antivibrio compounds produced by Pseudomonas sp W3: characterisation and assessment of their safety to shrimps Reviewed

    Pattamarat Rattanachuay, Duangporn Kantachote, Manee Tantirungkij, Teruhiko Nitoda, Hiroshi Kanzaki

    WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY   27 ( 4 )   869 - 880   2011.4

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    In order to explore compounds naturallly inhibitory to shrimp pathogenic vibrios, a culture filtrate of Pseudomonas sp. W3 at a pH of 2 was extracted with ethyl acetate (EtOAc) to produce 82.15 mg/l of a yellow-brown extract (EtOAc-W3) that had MIC values of 225-450 mu g/ml against the growth of 18 shrimp pathogenic Vibrio harveyi strains. The MIC of EtOAc-W3 against the most pathogenic strain PSU 2015 was 450 mu g/ml and this strain had the lowest LD(50) (50% lethal dose) to pacific white shrimp (Litopenaeus vannamei, PL 21). At this MIC value, EtOAc-W3 in artificial sea water (ASW) killed strain PSU 2015; however in natural sea water, only a partial growth inhibition was observed. The toxicity to pacific white shrimp and antivibrio activity of the EtOAc-W3 were investigated by conducting an experiment with 4 sets; native control (commercial ASW), EtOAc-W3 control (MIC/10, 45 mu g/ml), challenge (inoculation 6.0 x 10(6) c.f.u./ml PSU 2015) and treatment (6.0 x 10(6) c.f.u./ml PSU 2015 + 45 mu g/ml EtOAc-W3). The same experiment was repeated by increasing the dose of EtOAc-W3 to 90 mu g/ml (MIC/5). Both concentrations of EtOAc-W3 tested had no toxicity to postlarval shrimps. A significant decrease in shrimp mortality was observed over a 72 h period as approximately 80% of the shrimps died in each challenge set but only 63 and 23% died in the presence of 45 and 90 mu g/ml EtOAc-W3. The major component of EtOAc-W3 was supposed to be 2-heptyl-4-quinolone (HHQ) by FAB-MS and (1)H-NMR analyses of the purified fraction.

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  • MS/MS fragmentation-guided search of TMG-chitooligomycins and their structure-activity relationship in specific beta-N-acetylglucosaminidase inhibition

    Hirokazu Usuki, Yukihiro Yamamoto, Yuya Kumagai, Teruhiko Nitoda, Hiroshi Kanzaki, Tadashi Hatanaka

    ORGANIC & BIOMOLECULAR CHEMISTRY   9 ( 8 )   2943 - 2951   2011

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    The reducing tetrasaccharide TMG-chitotriomycin (1) is an inhibitor of beta-N-acetylglucosaminidase (GlcNAcase), produced by the actinomycete Streptomyces anulatus NBRC13369. The inhibitor shows a unique inhibitory spectrum, that is, selectivity toward enzymes from chitin-containing organisms such as insects and fungi. Nevertheless, its structure-selectivity relationship remains to be clarified. In this study, we conducted a structure-guided search of analogues of 1 in order to obtain diverse N,N,N-trimethylglucosaminium (TMG)-containing chitooligosaccharides. In this approach, the specific fragmentation profile of 1 on ESI-MS/MS analysis was used for the selective detection of desired compounds. As a result, two new analogues, named TMG-chitomonomycin (3) and TMG-chitobiomycin (2), were obtained from a culture filtrate of 1-producing Streptomyces. Their enzyme-inhibiting activity revealed that the potency and selectivity depended on the degree of polymerization of the reducing end GlcNAc units. Furthermore, a computational modeling study inspired the inhibitory mechanism of TMG-related compounds as a mimic of the substrate in the Michaelis complex of the GH20 enzyme. This study is an example of the successful application of a MS/MS experiment for structure-guided isolation of natural products.

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  • Characterization of the antiyeast compound and probiotic properties of a starter Lactobacillus plantarum DW3 for possible use in fermented plant beverages Reviewed

    Duangporn Kantachote, Pakorn Prachyakij, Wilawan Charernjiratrakul, Metta Ongsakul, Yodsawee Duangjitcharoen, Chaiyavat Chaiyasut, Teruhiko Nitoda, Hiroshi Kanzaki

    ELECTRONIC JOURNAL OF BIOTECHNOLOGY   13 ( 5 )   2010.9

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    Lactobacillus plantarum DW3 produced antifungal compounds that inhibited the growth of Rhodotorula mucilaginosa DKA, contaminating yeast in fermented plant beverages (FPBs) and various potential human pathogens. Phenyllactic acid (PLA) identified by gas chromatography- mass spectrometry (GC-MS) was produced at 31 mg/L PLA in MRS medium and 5 mg/ml inhibited growth of the target yeast in vitro by 90%. Other inhibitors were also present but not specifically identified. Results of in vitro tests showed that DW3 also had probiotic properties as it survived various human biological barriers resistance to pH 3, bile salts, growth without vitamin B(12) and the presence and absence of oxygen. Its inhibitory effect against foodborne pathogenic bacteria and spoilage organisms was higher than that found for a commercial strain Lactobacillus casei R. An acute oral toxicity test on ICR mice at a high single dose of either 10(9) and 10(12) cells per mouse for 14 days showed that DW3 had no adverse effect on the general health status and there was no evidence of bacteremia. Mice fed DW3 had a reduced weight gain compared to the control. No significant difference (p > 0.05) was found for the spleen weight index (SWI) among the treatment and control groups whereas there was a significant difference (p < 0.05) for the liver weight ratio (LWR) in a group fed with 10(12) cells per mouse when compared with the control group.

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  • Inhibition of shrimp pathogenic vibrios by extracellular compounds from a proteolytic bacterium Pseudomonas sp W3 Reviewed

    Pattamarat Rattanachuay, Duangporn Kantachote, Manee Tantirungkij, Teruhiko Nitoda, Hiroshi Kanzaki

    ELECTRONIC JOURNAL OF BIOTECHNOLOGY   13 ( 1 )   2010.1

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    Pseudomonas sp. W3, a bacterium known to produce an extracellular alkaline protease, secreted secondary metabolites that inhibited pathogenic bacteria responsible for shrimp luminous vibriosis disease. Antivibrio compounds in the culture supernatant or culture filtrates (0.45 mu m and 0.22 mu m) of the isolate W3 were tested using an agar well diffusion method on a number of pathogenic vibrios. Vibrio harveyi PSU 2015 a pathogenic isolate was the most sensitive strain. The effectiveness of preparations from the isolate W3 against V. harveyi PSU 2015, and V. cholerae PSSCMI 0062 was in the order of culture supernatant > 0.45 mu m culture filtrate > 0.22 mu m culture filtrate. These extracellular antivibrio compounds also lysed both dead and living cells of V. harveyi PSU 2015. Results of the partial characterization tests indicated that there was some particulate antivibrio compound that was destroyed by treatment with enzymes particularly achymotrypsin, autoclaving at 121 degrees C for 15 min and was mostly removed by filtration through a 0.22 mu m filter. Most of the inhibitory compounds were of small molecular weight able to pass through a 0.22 mu m filter and were resistant to treatment with various enzymes, pH values between 4-8 and temperatures up to 121 degrees C for 30 min. The optimum pH for the antivibrio activity in the 0.45 mu m culture filtrate was between pH 6-7.

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  • Pochonicine, a polyhydroxylated pyrrolizidine alkaloid from fungus Pochonia suchlasporia var. suchlasporia TAMA 87 as a potent beta-N-acetylglucosaminidase inhibitor Reviewed

    Hirokazu Usuki, Miho Toyo-oka, Hiroshi Kanzaki, Toru Okuda, Teruhiko Nitoda

    BIOORGANIC & MEDICINAL CHEMISTRY   17 ( 20 )   7248 - 7253   2009.10

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    A new polyhydroxylated pyrrolizidine alkaloid designated as pochonicine (1) was isolated from a solid fermentation culture of the fungal strain Pochonia suchlasporia var. suchlasporia TAMA 87. The structure of 1 was determined using NMR and MS techniques as (1R*, 3S*, 5S*, 6S*, 7R*, 7a S*)-5-acetamidomethyl-3-hydroxymethyl-1,6,7-trihydroxypyrrolizidine. Pochonicine (1) showed potent inhibition against beta-N-acetylglucosaminidases (GlcNAcases) of various organisms including insects, fungi, mammals, and a plant but no inhibition against beta-glucosidase of almond, alpha-glucosidase of yeast, or chitinase of Bacillus sp. The GlcNAcase inhibitory activity of pochonicine (1) was comparable to nagstatin, a potent GlcNAcase inhibitor of natural origin. (C) 2009 Elsevier Ltd. All rights reserved.

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  • TMG-chitotriomycin, an enzyme inhibitor specific for insect and fungal beta-N-acetylglucosaminidases, produced by actinomycete Streptomyces anulatus NBRC 13369 Reviewed

    Hirokazu Usuki, Teruhiko Nitoda, Misato Ichikawa, Nahoko Yamaji, Takashi Iwashita, Hajime Komura, Hiroshi Kanzaki

    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY   130 ( 12 )   4146 - 4152   2008.3

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    A novel beta-N-acetylglucosaminidase (GIcNAcase) inhibitor named TMG-chitotriomycin (1) was isolated from the culture filtrate of Streptomyces anulatus NBRC13369. The strain produced 1 only when colloidal chitin was used as the sole carbon source in the production medium. The structure of 1 was determined by spectral and constitutive sugar analyses of the corresponding alditol derivatives to be an equilibrated mixture of alpha-D-N,N,N-triMeGIcNH(2)-(1,4)-beta-D-GIcNAc-(1,4)-beta-D-GIcNAc-(1,4)-D-GlcNAc and its C-2 epimer of the reducing end residue. TMG-chitotriomycin (1) showed potent and selective inhibition of insect and fungal GIcNAcases with no inhibition of mammalian and plant GIcNAcases. In contrast, the known GIcNAcase inhibitor nagstatin potently inhibited all GIcNAcases. It should be emphasized that synthesized D-N,N,N-triMeGIcNH(2), which is the component sugar of 1, showed no inhibition of the insect Spodoptera litura GIcNAcase. These results suggest that the (GIcNAc)(3) unit positioned at the reducing end of 1 is essential for its enzyme inhibitory activity. The unique inhibitory spectrum of 1 will be useful to study chitinolytic systems and to develop selective fungicides or pesticides.

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  • Purification and characterization of a biosurfactant produced by Issatchenkia orientalis SR4 Reviewed

    Wichuda Katemai, Suppasil Maneerat, Fusako Kawai, Hiroshi Kanzaki, Teruhiko Nitoda, Aran H-Kittikun

    JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY   54 ( 1 )   79 - 82   2008.2

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  • Antinematodal activities of ingenane diterpenes from Euphorbia kansui and their derivatives against the pine wood nematode (Bursaphelenchus xylophilus) Reviewed

    Jianxiao Shi, Zhixuan Li, Teruhiko Nitoda, Minoru Izumi, Hiroshi Kanzaki, Naomichi Baba, Kazuyoshi Kawazu, Shuhei Nakajima

    ZEITSCHRIFT FUR NATURFORSCHUNG SECTION C-A JOURNAL OF BIOSCIENCES   63 ( 1-2 )   59 - 65   2008.1

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    Under the bioassay-guided method, two diterpenes, 3-O-(2",3"-dimethylbutanoyl)-13-O-dodecanoylingenol (1) and 3-O-(2",3"-dimethylbutanoyl)-13-O-decanoylingenol (2) isolated from Euphorbia kansui, showed a pronounced antinematodal activity against the nematode Bursaphelenchus xylophilus at the same minimum effective dose (MED) of 5 mu g per cotton ball and still displayed antinematodal activity at a dose of 2.5 mu g per cotton ball. Compounds 3-6 were obtained, and the structure of the new compound 6 was elucidated based on 1D- and 2D-NMR analyses and physicochemical data. Preliminary structure-biological activity relationships of ingenane-type compounds were deduced.

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  • Three antinematodal diterpenes from Euphorbia kansui Reviewed

    Jian-Xiao Shi, Zhi-Xuan Li, Teruhiko Nitoda, Minoru Izumi, Hiroshi Kanzaki, Naomichi Baba, Kazuyoshi Kawazu, Shuhei Nakajima

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   71 ( 4 )   1086 - 1089   2007.4

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    Three compounds, 20-O-acetyl-[3-O-(2'E,4'Z)-deca-dienoyl]-ingenol (1), 20-O-acetyl-[5-O-(2'E,4'Z)-decadienoyl]-ingenol (2) and 3-O-(2'E,4'Z)-decadienoylingenol (3), were isolated from Euphorbia kansui under the bioassay-guided method. Each compound showed the same antinematodal activity against the nematode, Bursaphelenchus xylophilus, at a minimum effective dose (MED) of 5 mu g/cotton ball.

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  • Chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste (Retracted Article. See vol 100, pg 6141, 2009) Reviewed

    Benjamas Thanomsub, Wanna Pumeechockchai, Anirut Limtrakul, Panarat Arunrattiyakorn, Wipawan Petchleelaha, Teruhiko Nitoda, Hiroshi Kanzaki

    BIORESOURCE TECHNOLOGY   98 ( 5 )   1149 - 1153   2007.3

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    The aim of this work was to study chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste. The culture produced two biosurfactants, a and b, which showed strong activity and were identified as L-rhamnopyranosyl-(L)-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate or Rha-Rha-C-10-C-10 and (L)-rhamnopyranosyl-(L)-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydodecanoate or Rha-Rha-C-10-C-12, respectively. Both compounds exhibited higher surfactant activities tested by the drop collapse test than several artificial surfactants such as SDS and Tween 80. Rhamnolipid a showed significant antiproliferative activity against human breast cancer cell line (MCF-7) at minimum inhibitory concentration (MIC) at 6.25 mu g/mL while rhamnolipid b showed MIC against insect cell line C6/36 at 50 mu g/mL. (c) 2006 Published by Elsevier Ltd.

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  • Enzymatic synthesis of dehydroderivatives from proline-containing cyclic dipeptides and their effects toward cell division Reviewed

    Panarat Arunrattiyakorn, Banri Ikeda, Teruhiko Nitoda, Hiroshi Kanzaki

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   71 ( 3 )   830 - 833   2007.3

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    We have previously isolated cyclo(L-PrO-L-Tyr) and cyclo(L-Phe-L-Pro) from an actinomycete by a novel enzymatic conversion-guided method. Their tetradehydro derivatives, cyclo(Delta Pro-Delta Tyr) and cyclo(Delta Phe-Delta Pro), were enzymatically prepared. Neither of them inhibited cell division, in contrast to other tetradehydro cyclic dipeptides prepared previously. This result suggests that an NH proton in a diketopiperazine ring and/or conformation of the compound are important for the activity.

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  • Chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste Reviewed

    Benjamas Thanomsub, Wanna Pumeechockchai, Anirut Limtrakul, Panarat Arunrattiyakorn, Wipawan Petchleelaha, Teruhiko Nitoda, Hiroshi Kanzaki

    BIORESOURCE TECHNOLOGY   97 ( 18 )   2457 - 2461   2006.12

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    The aim of this work was to study chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste. The culture produced two biosurfactants, a and b, which showed strong activity and were identified as L-rhamnopyranosyl-L-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate or Rha-Rha C-10-C-10 and L-rhamnopyranosyl-L-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydodecanoate or Rha-Rha C-10-C-12, respectively. Both compounds exhibited higher surfactant activities tested by the drop collapse test than several artificial surfactants such as SDS and Tween 80. Rhamnolipid a showed significant antiproliferative activity against human breast cancer cell line (MCF-7) at minimum inhibitory concentration (MIC) at 6.25 mu g/mL while rhamnolipid b showed MIC against insect cell line C6/36 at 50 mu g/mL. (c) 2005 Published by Elsevier Ltd.

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  • P-373 ISOLATION OF CYCLIC DIPEPTIDES FROM MICROBIAL CULTURE GUIDED BY ENZYMATIC CONVERSION

    Arunrattiyakorn Panarat, Nitoda Teruhiko, Kanzaki Hiroshi

    International Symposium on the Chemistry of Natural Products   2006   "P - 373"   2006.7

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    DOI: 10.24496/intnaturalprod.2006.0__P-373_

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  • P-374 ISOLATION AND STRUCTURE ELUCIDATION OF A NOVEL ENZYME INHIBITOR SPECIFIC FOR BETA-N-ACETYLGLUCOSAMINIDASES OF INSECT & FUNGAL ORIGIN

    Usuki Hirokazu, Nitoda Teruhiko, Yamaji Nahoko, Iwashita Takashi, Komura Hajime, Kanzaki Hiroshi

    International Symposium on the Chemistry of Natural Products   2006   "P - 374"   2006.7

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    DOI: 10.24496/intnaturalprod.2006.0__P-374_

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  • P-398 PRECURSOR-DIRECTED BIOSYNTHESIS & ENZYMATIC SYNTHESIS OF FLUORINATED ALBONOURSIN

    Kanzaki Hiroshi, Hirata Rie, Fukatani Yoshimi, Takeda Chikahiro, Ikeda Hajime, Nitoda Teruhiko

    International Symposium on the Chemistry of Natural Products   2006   "P - 398"   2006.7

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    DOI: 10.24496/intnaturalprod.2006.0__P-398_

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  • Enzymatic conversion-based method for screening cyclic dipeptide-producing microbes Reviewed

    P Arunrattiyakorn, T Nitoda, H Kanzaki

    PEPTIDES   27 ( 4 )   633 - 639   2006.4

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    We developed a method for screening cyclic dipeptide-producing microbes by enzymatic conversion. In this method, cyclic dipeptides are detected by the combination of: (i) conversion of cyclic dipeptides to dehydro cyclic dipeptides by cyclo(Leu-Phe) oxidase and (ii) detection of the dehydro derivative by UV spectrophotometry using TLC or HPLC analysis based on the absorbance change caused by the conversion. Using this method, the actinomycete strain A8 was isolated as a cyclic dipeptide-producing strain. The cyclic dipeptides were purified from the microbial extract by enzymatic detection-guided fractionation, and their structures were determined to be CYCIO(L-Phe-L-Pro) and cycIO(L-Pro-L-Tyr) by spectroscopic methods. (c) 2005 Elsevier Inc. All rights reserved.

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  • Activation of murine peritoneal macrophages by water-soluble extracts of Bursaphelenchus xylophilus, a pine wood nematode Reviewed

    H Kaji, A Tai, K Matsushita, H Kanzaki, Yamamoto, I

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   70 ( 1 )   203 - 210   2006.1

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    In our previous study, water-soluble extracts from Bursaphelenchus xylophilus (B. xylophilus), a pine wood nematode, were shown to enhance interleukin (IL)-4 plus lipopolysaccharide-induced polyclonal immunoglobulin (Ig) E production in vitro in mice and to increase serum levels of an antigen-nonspecific IgE in vivo. Here we examined whether the nematode extracts stimulate immunofunctions of murine peritoneal macrophages. In both resident and inflammatory macrophages, Fey receptor-mediated phagocytosis was markedly activated by B. xylophilus extracts, while non-specific phagocytosis was not. The enhancement of specific phagocytosis was accompanied by an increase in the formation of IgG-Fc gamma receptor rosettes. B. xylophilus extracts also stimulated IL-1 beta production in both types of macrophages, and enhanced NO production and mRNA expression of inflammatory cytokines in inflammatory macrophages. These results indicate that the extracts of B. xylophilus contain an activating substance(s) for immunofunctions in macrophages, besides an enhancing factor for polyclonal IgE production.

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  • Screening and partial characterization of inhibitors of insect beta-N-acetylglucosaminidase Reviewed

    H Usuki, T Nitoda, T Okuda, H Kanzaki

    JOURNAL OF PESTICIDE SCIENCE   31 ( 1 )   41 - 46   2006

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    Microbial culture broths were screened for novel beta-N-acetylglucosaminidase (GlcNAcase) inhibitors specific for the enzyme of insect origin. Four strains of actinomycetes, Streptomyces griseoloalbus JCM4480, S. clauifer JCM5059. S. anulatus NBRC13369 and S. griseus subsp. rhodochrous NBRC13849, produced unique compounds showing selective inhibition of the insect GlcNAcase. In contrast, 4 fungal strains, Paecilomyces sp. F13, F30, P. carneus F2281 and Verticillium sp. F40, were found to produce GlcNAcase inhibitors showing a broad spectrum of inhibitory activity against GlcNAcases from insects, mammals, plants and fungi. These results indicated obvious differences in GlcNAcases between insects and other organisms. This is the first report of enzyme inhibitors specific for the GlcNAcase of insect origin. (c) Pesticide Science Society of Japan.

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  • Bile acids are new products of a marine bacterium, Myroides sp strain SM1 Reviewed

    S Maneerat, T Nitoda, H Kanzaki, F Kawai

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   67 ( 5 )   679 - 683   2005.6

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    Strain SM1 was isolated as a biosurfactant-producing microorganism from seawater and presumptively identified as Myroides sp., based on morphology, biochemical characteristics and 16S rDNA sequence. The strain produced surface-active compounds in marine broth, which were purified, using emulsification activity for n-hexadecane as an indicator. The purified compounds were identified by thin-layer chromatography, H-1- and C-13-NMR spectra and fast atom bombardment mass spectrometry as cholic acid, deoxycholic acid and their glycine conjugates. Type strains of the genus Myroides, M. odoratus JCM7458 and M. odoramitimus JCM7460, also produced these compounds. Myroides sp. strain SM1 possessed a biosynthetic route to cholic acid from cholesterol. Thus, bile acids were found as new products of prokaryotic cells, genus Myroides.

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  • Enzymatic synthesis of dehydro cyclo(His-Phe)s, analogs of the potent cell cycle inhibitor, dehydrophenylahistin, and their inhibitory activities toward cell division Reviewed

    H Kanzaki, S Yanagisawa, T Nitoda

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   68 ( 11 )   2341 - 2345   2004.11

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    Cyclo(His-Phe) was effectively converted to its dehydro, derivatives by the enzyme of Streptomyces albulus KO-23, an albonoursin-producing actinomycete. Two types of dehydro derivatives were isolated from the reaction mixture and identified as cyclo(DeltaHis-DeltaPhe) and cyclo(His-DeltaPhe). This is the first report on cyclo(His-DeltaPhe) and the enzymatic preparation of both compounds. Cyclo(DeltaHis-DeltaPhe), a tetradehydro cyclic dipeptide, exhibited a minimum inhibitory concentration of 0.78 mumol/ml inhibitory activity toward the first cleavage of sea urchin embryos, in contrast to cyclo(His-APhe) that had no activity. The finding that the isoprenylated derivative of cyclo(DeltaHis-DeltaPhe), dehydrophyenylahistin, had 2,000 times higher activity than cyclo(DeltaHis-DeltaPhe) indicates that an isoprenyl group attached to an imidazole ring of the compound was essential for the inhibitory activity.

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  • Monoacylglycerols: glycolipid biosurfactants produced by a thermotolerant yeast, Candida ishiwadae Reviewed

    B Thanomsub, T Watcharachaipong, K Chotelersak, P Arunrattiyakorn, T Nitoda, H Kanzaki

    JOURNAL OF APPLIED MICROBIOLOGY   96 ( 3 )   588 - 592   2004

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    Aims: To isolate and characterize biosurfactants produced by a thermotolerant yeast isolated in Thailand.
    Materials and Results: Yeast strains isolated from plant material in Thailand were first screened for the ability to produce lipase and biosurfactant. A strain Y12, identified as Candida ishiwadae by physiological tests, survived at 45degreesC and produced relatively large amounts of biosurfactants. From the culture filtrate of this strain, two glycolipid biosurfactants, a and b, were purified by solvent fractionation, silica gel and ODS column chromatographies. Compounds a and b were determined to be monoacylglycerols; 1-linoleylglycerol and 1-oleylglycerol, respectively. Both compounds exhibited higher surfactant activities tested by the drop collapse test than several artificial surfactants such as sodium dodecyl sulphate.
    Conclusions: Glycolipid biosurfactants produced by a thermotolerant yeast, C. ishiwadae were characterized to be monoacylglycerols which exhibited high surfactant activities.
    Significance and Impact of the Study: A thermotolerant yeast strain, C. ishiwadae, could be a potential candidate for producing monoacylglycerols which are useful in industrial applications.

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  • A potent insect chitinase inhibitor of fungal origin Reviewed

    T Nitoda, H Usuki, H Kanzaki

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   58 ( 11-12 )   891 - 894   2003.11

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    A water-soluble polysaccharide was isolated from the culture filtrate of a fungal strain, Sphaeropsis sp. TNPT116-Cz, as a novel insect chitinase inhibitor. It was purified to chromatographic homogeneity by ethanol precipitation, anion-exchange and gel filtration chromatography. Its molecular weight was estimated to be 16 kDa by gel filtration HPLC. Monosaccharide analysis showed that it contained glucose, galactose, N-acetylglucosamine and a deoxysugar. This polysaccharide showed potent and specific inhibitory activity against Spodoptera litura chitinase with an IC50 value of 28 nm.

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  • Augmentation by Bursaphelenchus xylophilus, a pine wood nematode, of polyclonal IgE production induced by lipopolysaccharide plus interleukin-4 in murine splenocytes Reviewed

    H Kaji, M Kawada, A Tai, H Kanzaki, Yamamoto, I

    JOURNAL OF PHARMACOLOGICAL SCIENCES   91 ( 2 )   158 - 162   2003.2

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    Bursaphelenchus xylophilus (B. xylophilus) is a pine wood nematode that is known to cause pine wilt disease. We report here that B. xylophilus extracts augmented the polyclonal immunoglobulin E (IgE) production induced by lipopolysaccharide (LPS) plus interleukin-4 (IL-4) both in murine splenocytes and purified B cells as determined by ELISA and ELIspot assays, but they did not cause such a promotion in the absence of either LPS or IL-4. We also observed that the antigen-nonspecific IgE levels were increased in sera of mice treated with B. xylophilus extracts, which were comparable to those of Ascaris suum extracts. These findings suggest that administration of B. xylophilus extracts could suppress allergic diseases via a saturation of mast cell Fcepsilon receptors or/and an inhibition of antigen-specific IgE synthesis to the allergen by a polyclonal response.

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  • Effective Production of Potent Cell Cycle Inhibitor Dehydrophenylahistin by a Combination of Chemical Racemization and Streptomyces Enzyme-catalyzed Conversion. Reviewed

    Hiroshi Kanzaki, Banri Ikeda, Teruhiko Nitoda

    Actinomycetologica   17 ( 1 )   1 - 5   2003

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  • Macromolecular insect chitinase inhibitors produced by fungi: Screening and partial characterization Reviewed

    T Nitoda, H Usuki, A Kurata, H Kanzaki

    JOURNAL OF PESTICIDE SCIENCE   28 ( 1 )   33 - 36   2003

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    Culture broths of fungal strains were screened for novel insect chitinase inhibitors using the Spodoptera litura chitinase inhibitory assay. The culture filtrates of 5 strains showed potent and specific inhibitory activity against the insect chitinase. Partial characterization showed that the active compounds produced by these strains were water-soluble macromolecular compounds which had not been hitherto reported as chitinase inhibitors. These novel chitinase inhibitors are, therefore, expected to be potential agents for insect control.

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  • A novel potent cell cycle inhibitor dehydrophenylahistin - Enzymatic synthesis and inhibitory activity toward sea urchin embryo Reviewed

    H Kanzaki, S Yanagisawa, K Kanoh, T Nitoda

    JOURNAL OF ANTIBIOTICS   55 ( 12 )   1042 - 1047   2002.12

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    A novel dehydrogenated cyclic dipeptide named as dehydrophenylahistin (DeltaPLH) was effectively prepared from a fungal metabolite (-)-phenylahistin by the enzymatic conversion catalyzed by the cell-free extract of Streptomyces albulus KO-23, an albonoursin-producing actinomycete. DeltaPLH exhibited more than 1,000 times as high potent inhibitory activity toward the first cleavage of sea urchin embryos as (-)-phenylahisitn which has been reported to be a cell cycle inhibitor and more than 10,000 as high as albonoursin, indicating that DeltaPLH is a promising leading compound for anticancer drugs.

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  • Biosynthetic intermediates of the tetradehydro cyclic dipeptide albonoursin produced by Streptomyces albulus KO-23 Reviewed

    H Kanzaki, S Yanagisawa, T Nitoda

    JOURNAL OF ANTIBIOTICS   53 ( 11 )   1257 - 1264   2000.11

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    The cell-free extract of an albonoursin-producing strain Streptomyces albulus KO-23 catalyzes the conversion of cyclo(L-Leu-L-Phe) (1) to albonoursin (2). At the early stage of this conversion, two compounds were newly formed prior to albonoursin synthesis in the reaction mixture. These compounds were isolated and identified as (Z)-3-benzylidene-6-isobutyl-2,5-piperazinedione (4) and (Z)-3-benzyl-6-isobutylidene-2,5-piperazinedione (3). The cell-free extract also catalyzed the conversion of compound 3 or 4 to albonoursin. From these results, albonoursin was found to be biosynthesized via these compounds from cyclo(L-Leu-L-Phe). These didehydro diketopiperazines exhibited no inhibitory activity toward the first cleavage of sea urchin embryo in contrast to the higher cytotoxicity for albonoursin, indicating that dehydrogenation at alpha,beta -positions of both amino acid residues in diketopiperazines is required for cytotoxicity.

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  • Novel diketopiperazine metabolism in a microorganism: Two-step hydrolysis of cyclo(Gly-Leu) to amino acids and preliminary characterization of cyclo(Gly-Leu) hydrolase and dipeptidase Reviewed

    H Kanzaki, N Mizuta, T Nitoda, K Kawazu

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   89 ( 6 )   602 - 605   2000.6

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    A bacterium, strain NM 5-3, isolated from soil exhibited the highest cyclo(Gly-Leu) (CGL)-hydrolyzing activity and was identified as Agrobacterium radiobacter. The reaction products from CGL were dipeptides (Leu-Gly and Gly-Leu) and amino acids (Leu and Gly). Inhibitors for the dipeptidase of this strain did not inhibit the hydrolysis of CGL to dipeptides, indicating that two distinct enzymes, CGLase and a dipeptidase, were involved in its hydrolysis. The activities of these two enzymes were separated by anion-exchange column chromatography. The results indicated that strain NM5-3 hydrolyzed CGL via the dipeptides to the corresponding amino acids. The CGLase fraction was found to catalyze the hydrolysis of cyclo(Gly-D-Leu), cyclo(Gly-Gly), cyclo(L-Ala-Gly), and cyclo(D-Ala-Gly). On the other hand, the dipeptidase fraction exhibited L-specific substrate specificity.

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  • Two antinematodal phenolics from Knema hookeriana, a Sumatran rainforest plant Reviewed

    Y Alen, S Nakajima, T Nitoda, N Baba, H Kanzaki, K Kawazu

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   55 ( 3-4 )   300 - 303   2000.3

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    The activity-guided chromatographic purification of the methanol extract of Knema hookeriana, using pine wood nematodes Bursaphelenchus xylophilus has successfully led to the isolation and characterization of two phenolic antinematodal compounds with minimun effective dose (MED) of 4.5 and 20 mu g/cotton ball (mu g/ bl.) or 0.018 and 0.073 mu M/cotton ball (mu M/bl.), respectively. Based on their chemical and spectral properties, these compounds were determined to be 3-undecylphenol (1) and 3-(8Z-tridecenyl)-phenol (2). These compounds were isolated for the first time from this species, and 2 seems to be a novel compound.

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  • Antinematodal activity of some tropical rainforest plants against the pinewood nematode, Bursaphelenchus xylophilus Reviewed

    Y Alen, S Nakajima, T Nitoda, N Baba, H Kanzaki, K Kawazu

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   55 ( 3-4 )   295 - 299   2000.3

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    Sixty five methanolic extracts of Sumatran rainforest plants representing 63 species of 21 families were assayed in vivo for antinematodal activity against Bursaphelenchus xylophilus using our cotton ball-fungal mat method. Extracts of 27 plants species from 14 families exhibited antinematodal activity, while 37 species were inactive. Among them, three extracts of Bischofia javanica, Knema hookeriana and Areca catechu exhibited very strong activity at minimum effective dose (MED) of 0.7 mg/cotton ball (mg/bl.). Eight extracts from Allamanda cathartica, Ervatamia corymbosa, Hoya diversifolia, Bischofia javanica, Derris malacensis, Melastoma melabathricum, Ophiorriza konsteleary and Brucea sumatrana also showed strong activity (MED, 5 mg/bl.).

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  • Enzymatic conversion of cyclic dipeptides to dehydro derivatives that inhibit cell division Reviewed

    Hiroshi Kanzaki, Daisuke Imura, Teruhiko Nitoda, Kazuyoshi Kawazu

    Journal of Bioscience and Bioengineering   90 ( 1 )   86 - 89   2000

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    The cell-free extract of an albonoursin-producing strain, Streptomyces albulus KO-23, was found to catalyze the conversion of several cyclic dipeptides having Phe and aliphatic side chain-containing amino acid residues to the corresponding dehydro derivatives. 3Z-Benzylidene-6S-methyl-2,5-piperazinedione, 3Z-benzylidene-2,5-piperazinedione, and 3Z, 6Z-dibenzylidene-2,5-piperazinedione were prepared by this conversion system. Among the dehydro cyclic dipeptides prepared, tetradehydro derivatives exhibited inhibitory activity toward the first cleavage of sea urchin embryo, while didehydro derivatives did not. We previously found that cyclo(Leu-Phe) and its didehydro derivatives did not show any inhibitory activity, in contrast to high activity in the case of albonoursin. Taken together, these findings indicate that dehydrogenation at the α, β-positions of both amino acid residues in this type of cyclic dipeptide is required for the inhibitory activity.

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  • Nostofungicidine, an antifungal lipopeptide from the field-grown terrestrial blue-green alga Nostoc commune

    S Kajiyama, H Kanzaki, K Kawazu, A Kobayashi

    TETRAHEDRON LETTERS   39 ( 22 )   3737 - 3740   1998.5

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    In the course of our screening program for bioactive compounds, a novel lipopeptide, nostofungicidine (1), was isolated from the methanolic extract of a field-grown terrestrial blue-green alga, Nostoc commune. The structure of nostofungicidine was elucidated by chemical degradation and extensive NMR measurements including DQF-COSY, HOHAHA, HMBC, and ROESY techniques. Nostofungicidine contains a novel beta-amino acid 3-amino-6-hydroxy stearic acid (Ahs) in its structure. (C) 1998 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0040-4039(98)00573-5

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  • Novel bioactive oxazolomycin isomers produced by Streptomyces albus JA3453 Reviewed

    H Kanzaki, K Wada, T Nitoda, K Kawazu

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   62 ( 3 )   438 - 442   1998.3

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    Two novel oxazolomycin isomers, oxazolomycins B (2) and C (3), were isolated from the fermentation broth of an oxazolomycin-producing strain, Streptomyces albus JA3453. Both compounds are geometrical isomers of oxazolomycin (1), the configurations of their triene moieties being (4'E, 6'E, 8'E) (2) and (4'Z, 6'E, 8'E) (3) while that of oxazolomycin fl) is (4'Z, 6'Z, 8'E), Compounds 2 and 3 exhibited potent inhibitory activity against crown gall formation with the same MIC (0.8 mu g/disk) as oxazolomycin. Compounds 2 and 3 showed no antibacterial activity against Agrobacterium tumefaciens, in contrast to oxazolomycin which has specific anti-A. tumefaciens activity.

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  • Antimicrobial constituents of Angelica dahurica roots

    YS Kwon, A Kobayashi, SI Kajiyama, K Kawazu, H Kanzaki, CM Kim

    PHYTOCHEMISTRY   44 ( 5 )   887 - 889   1997.3

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    One novel coumarin from Angelica dahurica roots was elucidated to be 5,8-di(2,3-dihydroxy-3-methylbutoxy)-psoralen. It occurs together with six other known coumarins and ferulic acid. The antimicrobial activity of the coumarins and ferulic acid were compared. Copyright (C) 1997 Elsevier Science Ltd.

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  • Accumulation of Benzoic Acid in Suspension Cultured Cells of Pinus thunbergii Parl. in Response to Phenylacetic Acid Administration

    KAWAZU Kazuyoshi, ZHANG Hong, KANZAKI Hiroshi

    Bioscience, biotechnology, and biochemistry   60 ( 9 )   1410 - 1412   1996.9

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    The generation and accumulation of both benzoic acid (BA) and its conjugates were induced in suspension cultured cells of Pinus thunbergii by administering either phenylacetic acid (PA), a toxic metabolite of Bacillus cereus (strain HY-3) accompanying the pine wood nematode, or a lyophilized culture supernatant of this bacterium. BA conjugates reached their maximal levels in quantity two days after the administration and then decreased gradually until the 14th day, while BA increased significantly throughout this period. This pattern is similar to that in 3-year-old pine trees treated with PA, suggesting that the pathological reaction of pine tissues to the PA toxin might be involved in the pathogenesis mechanism for the pine wilt disease.

    DOI: 10.1271/bbb.60.1410

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  • Relationship between the Pathogenicity of the Pine Wood Nematode, Bursaphelenchus xylophilus, and Phenylacetic Acid Production

    KAWAZU Kazuyoshi, ZHANG Hong, YAMASHITA Hideaki, KANZAKI Hiroshi

    Bioscience, biotechnology, and biochemistry   60 ( 9 )   1413 - 1415   1996.9

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    Phenylacetic acid (PA), a toxin produced by three strains of bacteria accompanying the pine wood nematode, Bursaphelenchus xylophilus, was found to be formed in a culture of the nematode. An animal nutrient, nutrient broth (NB) medium, was more suitable for PA production of the accompanying bacteria than a vegetable nutrient, potato sucrose malt extract (PSM) medium. It is presumed that dead nematodes in the PSM medium provided the bacteria with nutrient for PA production. In the culture of virulent isolate OKD-3, more PA was detected than in that of less-virulent isolate OKD-1. PA production of the accompanying bacteria can dominate the pathogenicity of the nematode.

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  • A cytotoxic principle of Tamarindus indica, di-n-butyl malate and the structure-activity relationship of its analogues

    A Kobayashi, MI Adenan, SI Kajiyama, H Kanzaki, K Kawazu

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   51 ( 3-4 )   233 - 242   1996.3

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    Bioassay-guided fractionation of the methanolic extract of Tamarindus indica fruits led to the isolation of L-(-)-di-n-butyl malate which exhibited a pronounced cytotoxic activity against sea urchin embryo cells. In order to study structure-activity relationships, close-structure relatives of di-n-butyl malate were synthesized using D-(+)- and L-(-)-malic acid as starting materials, and their cytotoxic activities were examined for the sea urchin embryo assay. L-(-)-Di-n-pentyl malate was the most effective inhibitor to the development of the fertilized eggs. Significant inhibitory activity was not seen in the esters of D-(-)-isomer.

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  • ANTIFUNGAL COMPOUNDS INDUCED IN THE DUAL CULTURE WITH PHYTOLACCA-AMERICANA CALLUS AND BOTRYTIS-FABAE

    A KOBAYASHI, K HAGIHARA, S KAJIYAMA, H KANZAKI, K KAWAZU

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   50 ( 5-6 )   398 - 402   1995.5

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    In order to investigate new metabolites which are only induced in a plant callus infected by a pathogenic fungus, dual cultures with combinations of 10 species of fungi and 6 plant cell lines from different species were established. Among the combinations tested, the methanolic extract of a dual culture consisting of a plant cell line, Phytolacca americana and a fungus, Botrytis fabae showed a marked antifungal activity to Cladosporium herbarum. The main active constituent of this extract was identified to be phytolaccoside B (Pls B) by the spectroscopic analyses.

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  • A NEW-TYPE OF ANTIMICROBIAL PHENOLICS PRODUCED BY PLANT PEROXIDASE AND ITS POSSIBLE ROLE IN THE CHEMICAL DEFENSE SYSTEMS AGAINST PLANT-PATHOGENS

    A KOBAYASHI, Y KOGUCHI, H KANZAKI, SI KAJIYAMA, K KAWAZU

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   49 ( 7-8 )   411 - 414   1994.7

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    Syringaldehyde readily reacted in the horse-radish peroxidase (HRPOD) system. The ethyl acetate extract of the reaction mixture showed a marked antimicrobial activity against bacteria and fungi. After repeated column chromatography three potential antimicrobial compounds were obtained from the extract. The structural elucidation of active compounds was achieved by a combination of spectroscopic techniques and chemical modification.

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  • NOSTODIONE-A, A NOVEL MITOTIC SPINDLE POISON FROM A BLUE-GREEN-ALGA NOSTOC-COMMUNE

    A KOBAYASHI, SI KAJIYAMA, K INAWAKA, H KANZAKI, K KAWAZU

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   49 ( 7-8 )   464 - 470   1994.7

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    A novel antimitotic compound named Nostodione A (Nd A) was isolated from a terrestrial blue-green alga Nostoc commune. Nostodione A disturbed the mitotic spindle formation of sea-urchin eggs and gave small spindles with low birefringence density. Nostodione A, however, had no phytotoxicity on the germination of a dicotyledonous plant Medicago sativa. Based on the spectral analysis and chemical degradation, the structure of Nostodione A was elucidated.

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  • PRODUCTION OF A NEW-TYPE OF BIOACTIVE PHENOLIC COMPOUND

    A KOBAYASHI, Y KOGUCHI, H KANZAKI, S KAJIYAMA, K KAWAZU

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   58 ( 1 )   133 - 134   1994.1

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    Peroxidase (POD) catalyzed the polymerization of naturally occurring phenolics to give a variety of bioactive products. The guaiacol readily reacted in the POD system and afforded potential anti-microbial compounds. Three active phenols, 1, 2, and 3, were isolated from the reaction mixture. Spectroscopic analyses elucidated that 1 and 2 were a dimer and a trimer respectively originating from guaiacol. Compound 3 was readily oxidized to 4 by exposure to air. The structure of 4 was determined by a combination of chemical modification and spectroscopic analyses, and 3 was elucidated to be the hydroxyquinone form of 4.

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  • ELICITOR-ACTIVE OLIGOSACCHARIDES FROM ALGAL LAMINARAN STIMULATE THE PRODUCTION OF ANTIFUNGAL COMPOUNDS IN ALFALFA Reviewed

    A KOBAYASHI, A TAI, H KANZAKI, K KAWAZU

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   48 ( 7-8 )   575 - 579   1993.7

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    Natural polysaccharides were examined for their activity as elicitors of phytoalexins and flavonoids in alfalfa cotyledons. Only two polysaccharides, laminaran and pectic acid, had elicitor activity. Laminaran, which was more active than pectic acid, was hydrolyzed by tunicase and the hydrolysate was subjected to charcoal and gel filtration columns. Introduction of the pyridylamino group into the elicitor-active oligosaccharides was attempted in order to facilitate isolation. The pyridylamino derivatives were found to exhibit higher activity than the original oligosaccharides. Their liquid chromatography mass spectrometry (LC-MS) analysis revealed that the elicitor-active principles form two ion clusters with the same molecular weights, m/z 1070 and 1232. Their high performance liquid chromatography (HPLC) analysis showed three main peaks. The individual peaks (LN-1, 2,3) were collected and subjected to the alfalfa cotyledon assay. LN-3 showed the highest activity (minimum effective concentration, 0.8 mug/ml).

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  • ST-1, a Novel Radical Scavenger From Fungus F-124

    Kobayashi Akio, Koguchi Yutaka, Takahashi Shigeo, Kanzaki Hiroshi, Kawazu Kazuyoshi

    Bioscience, biotechnology, and biochemistry   57 ( 6 )   1034 - 1036   1993.6

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    Peroxidase (POD) plays an important role in the biosynthesis of disease-lignin, which protects plants from fungal invasion. A novel POD inhibitor, ST-1 (1), was isolated from a fungus, and its chemical structure was elucidated to be 1,3-dihydro-4,5,6-trihydroxy-1,3-dimethoxy-7-methylisobenzofuran by a combination of chemical modification and spectroscopic techniques. ST-1 effectively scavenged oxygen radicals and also suppressed the phytoalexin accumulation in kidney bean.

    DOI: 10.1271/bbb.57.1034

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  • Bioconversion of olive leaf constituents by Aspergillus solid state fermentation using an air flow type rotary solid state fermenter for laboratory use

    辰巳七海, 橋本敦子, 奥川日菜乃, 三宅剛史, 伊藤一成, 谷野有佳, 竹内赴登, 山下秀行, 内田真美, 三木翔平, 吉田靖弘, 徐恵美, 菊地敬一, 深野夏暉, 仁戸田照彦, 仁戸田照彦, 神崎浩, 神崎浩

    日本農芸化学会大会講演要旨集(Web)   2023   2023

  • Bioconversion of skin and seed constituents by Aspergillus solid state fermentation of wine pomace

    藤田彩楓, 奥川日菜乃, 橋本敦子, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 中川拓郎, 平野幸司, 仁戸田照彦, 仁戸田照彦, 神崎浩, 神崎浩

    日本農芸化学会大会講演要旨集(Web)   2023   2023

  • Creation of highly functionalized Japanese wine pomace koji using koji mold solid culture technology

    神崎浩, 仁戸田照彦, 伊藤一成, 谷野有佳, 竹内赴登

    Annual Report of the Okayama Foundation for Science and Technology   32 ( 32 )   26 - 30   2023

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  • バイオサーファクタント・サーファクチンの自己集合特性への対カチオン種の違いによる影響

    柳澤恵広, 柳澤恵広, 平敏彰, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)   62nd   2022

  • Effect of hot water pretreatment on Aspergillus solid state fermentation of wine pomace

    奥川日菜乃, 橋本敦子, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 中川拓郎, 平野幸司, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)   2022   2022

  • Cyclo(Leu-Phe)oxidaseの幅広い基質特異性の定量的評価-天然の電子受容体を用いた活性測定と人工の電子受容体を用いた活性測定の比較-

    坂口幸士朗, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)   61st   2022

  • Analysis of MS/MS fragmentation patterns for search of new asteltoxin analogs with a unique α-pyrone moiety

    三谷尚大, 神崎浩, 仁戸田照彦

    日本農薬学会大会講演要旨集   47th (CD-ROM)   2022

  • Effect of steaming on Aspergillus solid state fermentation of olive leaves

    橋本敦子, 奥川日菜乃, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 内田真美, 三木翔平, 吉田靖弘, 徐恵美, 菊地敬一, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)   2022   2022

  • 麹菌固体培養による日本ワインパミス成分の高機能化(第2報)

    奥川日菜乃, 橋本敦子, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 中川拓郎, 平野幸司, 仁戸田照彦, 神崎浩

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集   2021 (CD-ROM)   2021

  • Quantitative evaluation of the substrate specificity of cyclo(Leu-Phe)oxidase toward various cyclodipeptides

    坂口幸士朗, 小西健太, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集   22nd (CD-ROM)   2020

  • 地方発バイオイノベーションの進展と機能性食素材開発 岡山発,産学連携フードイノベーション

    神崎浩

    Food Style 21   21 ( 1 )   2017

  • 地方発バイオイノベーションの進展と機能性食素材開発 岡山発,産学連携フードイノベーション 2)

    神崎浩

    Food Style 21   21 ( 2 )   2017

  • MS/MSフラグメンテーションプロファイルを指標としたTMG-chitotriomycin類縁体の探索と構造活性相関

    神崎浩, 臼木博一, 臼木博一, 仁戸田照彦, 畑中唯史

    日本農薬学会大会講演要旨集   36th   2011

  • Conversion of secondary metabolites from olive leaves by Baker's yeasts and the antioxidative activity of the product

    Erika Akihisa, Yoshihiro Harada, Kumi Kobayashi, Akiko Hirano, Teruhiko Nitoda, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   62 ( 1 )   123 - 123   2010.1

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  • 生物種の由来が異なる酵素を見分ける阻害剤 新規な構造と活性をもつ放線菌由来β-N-アセチルグルコサミニダーゼ阻害剤

    仁戸田 照彦, 臼木 博一, 神崎 浩

    化学と生物   47 ( 3 )   158 - 160   2009

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    DOI: 10.1271/kagakutoseibutsu.47.158

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  • Bioassays: inhibitors of insect chitin-degrading enzymes

    Teruhiko Nitoda, Hiroshi Kanzaki

    Isolation, Identification and Characterization of Allelochemicals/Natural Products   499 - 412   2009

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  • 岡山大産米100%使用,日本酒「おお岡大」― 学産学消から学産地消を目指して ―

    神崎浩

    日本生物工学会誌   86 ( 5 )   250 - 251   2008

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  • Development of natural cosmetic materials-microbial conversion of olive antioxidative constituents

    Fragrance journal   36 ( 2 )   63 - 66   2008

  • Purification and characterization of cyclo (Leu-Phe) oxidase of Streptomyces albulus expressed in Escherichia coli

    Machiko Nagao, Rie Hirata, Teruhiko Nitoda, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   48 ( 3-4 )   108 - 108   2007.9

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  • A novel β-N-acetylglucosaminidase inhibitor of fungal origin

    NITODA Teruhiko, TOYOOKA Miho, USUKI Hirokazu, OKUDA Toru, KANZAKI Hiroshi

    32   123 - 123   2007.3

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  • Microorganisms with homocysteine thiolactone hydrolase: Screening and stereospecificity - Abstracts

    Shozo Honda, Teruhiko Nitoda, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   42 ( 3-4 )   126 - 127   2006.11

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  • Further proof for wide substrate specificity of cyclo(Leu-Phe) oxidase from an actinomycete: Enzymatic conversion of cyclic dipeptides containing non-aromatic amino acids - Abstracts

    Rie Hirata, Teruhiko Nitoda, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   42 ( 3-4 )   127 - 128   2006.11

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  • Cloning and expression of cyclo(Leu-Phe) oxidase gene from an actinomycete Streptomyces albulus K023

    Machiko Nagao, Atsushi Morimoto, Takashi Tamura, Tohru Dairi, Hiroshi Kanzaki

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   42 ( 3-4 )   126 - 126   2006.11

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  • Inhibitory specificity of a novel β-N-acetylglucosaminidase inhibitor HU-1

    USUKI Hirokazu, NITODA Teruhiko, KANZAKI Hiroshi

    31   81 - 81   2006.3

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  • Structure elucidation of a novel β-N-acetylglucosaminidase inhibitor HU-1

    USUKI Hirokazu, NITODA Teruhiko, YAMAJI Nahoko, IWASHITA Takashi, KOMURA Hajime, KANZAKI Hiroshi

    31   82 - 82   2006.3

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  • 抗がん活性化合物の酵素合成

    神崎 浩

    ケミカルエンジニアリング   50, 902-906   2005

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  • 2D15-5 Synthesis of novel fluoro analogs of albonoursin by precursor-directed biosynthesis

    KANZAKI Hiroshi, TAKEDA Chikahiro, FUKATANI Yoshimi, NITODA Teruhiko

    17   135 - 135   2005

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  • 2J13-4 Thionation of a cell cycle inhibitor dehydrophenylahistin

    KANZAKI Hiroshi, IKEDA Banri, TAMURA Takashi, NITODA Teruhiko

    16   237 - 237   2004

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  • デヒドロアミノ酸残基を含む生理活性環状ジペプチド類

    神崎 浩

    生物工学会誌   81, 396-399 ( 9 )   396 - 399   2003

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  • Enzymatic synthesis of a cell cycle inhibitor dehydrophenylahistin

    Kanzaki Hiroshi, Ikeda Banri, Morimoto Atsushi, Nitoda Teruhiko

    15   60 - 60   2003

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  • Chemical information-guided enzymatic synthesis of bioactive compounds

    KANZAKI Hiroshi

    Nippon Nōgeikagaku Kaishi   76, 539-541 ( 6 )   539 - 541   2002

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    DOI: 10.1271/nogeikagaku1924.76.539

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  • 環状ジペプチド脱水素酵素で新規生理活性物質を作る

    神崎 浩

    バイオサイエンスとインダストリー   60, 26-29   2002

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  • 649 Purification and characterization of dipeptidase from a cyclic dipeptide-assimilating bacterium

    Kanzaki Hiroshi, Miyoshi Kazuyuki, Nitoda Teruhiko

    14   107 - 107   2002

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  • 1 Production of Bioactive Compounds by Biosynthetic Enzymes of Dehydro Cyclic Dipeptides

    Kanzaki Hiroshi, Yanagisawa Satohiro, Akazawa Kazumi, Ikeda Banri, Morimoto Atsushi, Nitoda Teruhiko

    ( 43 )   1 - 5   2001.9

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    Cyclic dipeptides (CDPs, diketopiperazines) and their derivatives are widely distributed in nature as secondary metabolites. Although some of dehydro-CDPs are known as cell cycle inhibitors, their effective syntheses have not been reported. We found that Streptomyces albulus KO23, an albonoursin-producing actinomycete, had a biosynthetic pathway from cyclo (Leu-Phe) to albonoursin, cyclo (ΔLeu-ΔPhe) by the fed-batch culture and the resting-cell experiments. And this enzyme activity was found to be effectively extracted in the cell-free extract of this actinomycete. This is the first report for the dehydrogenation of amino acid residues at α,β-positions in CDPs. Furthermore, this enzyme system enables us to synthesize several didehydro- and tetradehydro-CDPs from the corresponding CDPs. Among dehydro-CDPs prepared, the tetradehydro-CDPs exhibited cytotoxicity, while the didehydro-CDPs had no activity, indicating that dehydrogenation at α,β-positions of both amino acid residues in CDPs is required for cytotoxicity. Based on the above results, we speculated that a tetradehydro-CDP prepared from a didehydro-CDP exhibiting cytotoxicity might be a potent cytotoxic compound. Dehydrophenylahistin synthesized by this enzyme system from (-)-phenylahistin, which was recently reported to be a new cell cycle inhibitor, exhibited 1000 times higher inhibitory activity toward the first cleavage of sea urchin embryo than (-)-phenylahistin, and thus, would be a promising lead compound for antitumor agents.

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  • Production of Bioactive Compounds by Microbial Diketopiperazine-Metabolizing Enzymes

    KANZAKI HIROSHI

    79(3), 71-77 ( 3 )   71 - 77   2001

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    iketopiperazines(DKPs)and their derivatives are widely distributed in nature as secondary metabolites.Although some dehydroDKPs are known as cell cycle inhibitors, their effective synthesis has not been reported. We found that the cell-free extract from an albonoursin-producing actinomycete, Streptomyces albulus KO23, catalyzed the conversion of cyclo(Leu-Phe)to albonoursin. This is the first report on the dehydrogenation of amino acid residues at the α, β-positions in DKPs. Furthermore, this enzyme system enabled us to synthesize several didehydro- and tetradehydro- DKPs from the corresponding DKPs. Among the dehydroDKPs prepared, dehydrophenylahistin from (-)-phenylahistin, which was recently reported to be a new cell cycle inhibitor, exhibited 1000 times higher inhibitory activity toward the first clevage of sea urchin embrvo than (-)-phenylahistin, and would thus be a promising lead compound for antitumor agents. Agrobacterium radiobacter NM 5-3 isolated from soil hydrolyzed CGL to from dipeptides(i.e., Leu-Gly and Gly-Leu)and amino acids(i.e., Leu and Gly). This CGL hydrolysis was catalyzed by two distinct enzymes, CGLase and dipeptidase, which were separated by anion-exchange column chromatography. The CGLase was found to catalyze the hydrolysis of CGDL, CGG, CAG, and CDAG. On the other hand, the dipeptidase exhibited L-specific substrate specificity.

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  • Effective production of dehydro cyclic dipeptide albonoursin exhibiting pronuclear fusion inhibitory activity II. Biosynthetic and bioconversion studies

    Hiroshi Kanzaki, Daisuke Imura, Teruhiko Nitoda, Kazuyoshi Kawazu

    Journal of Antibiotics   53 ( 1 )   58 - 62   2000

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    Albornoursin production was greatly enhanced when cycle (r-Leu-L-Phe) (CFL), a tetrahydro derivative of albonoursin, was added to the 2-day culture of an albonoursin-producing actinomycete, Streptomyces albulus KO-23. The increase in albonoursin production paralleled the amount of CFL added. Furthermore, the resting cells of the strain catalyzed the bioconversion of CFL to albonoursin. The optimum pH and temperature for the conversion were found to be pH 10.0 and 50°C. The feeding experiments and the resting-cell reactions revealed that albonoursin is biosynthesized by dehydrogenation of CFL in the actinomycete. This is the first report for a dehydrogenation of amino acid residues at the α,β-positions in cyclic dipeptides.

    DOI: 10.7164/antibiotics.53.58

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  • A novel microbial enzyme system for hydrolyzing a cyclic dipeptide, cyclo(Gly-Leu)

    Kanzaki Hiroshi, Miyoshi Kazuyuki, Mizuta Naoki, Nitoda Teruhiko

    12   210 - 210   2000

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  • Improved bioassay method for Spodoptera litura chitinase inhibitors using a colloidal chitin powder with a uniform particle size as substrate

    T Nitoda, H Kurumatani, H Kanzaki, K Kawazu

    PESTICIDE SCIENCE   55 ( 5 )   563 - 565   1999.5

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    A previously reported bioassay method for Spodoptera litura chitinase inhibitors has been improved by use of colloidal chitin powder with a uniform particle size. This improvement made the assay four times more sensitive. Detection of three active supernatants by screening of supernatants and cell extracts from 135 fermentation broths has proved the efficiency of this improved method. (C) 1999 Society of Chemical Industry.

    DOI: 10.1002/(SICI)1096-9063(199905)55:5<563::AID-PS972>3.0.CO;2-#

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  • Enzymatic dehydrogenation of cyclo(L-Phe-L-Leu) to a bioactive derivative, albonoursin

    H Kanzaki, D Imura, T Nitoda, K Kawazu

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   6 ( 3 )   265 - 270   1999.3

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    The cell-free extract of Streptomyces sp. KO-2388, an albonoursin-producing strain, was found to catalyze the conversion of cyclo(L-Phe-L-Leu) to albonoursin. The conversion activity was simply determined by measuring the increase in ultraviolet (UV) absorption of the reaction mixture at 317 nm, lambda(max) (epsilon 25,400) of albonoursin, where cyclo(L-Phe-L-Leu) had no absorption. The optimum pH and temperature of this bioconversion using the cell-free extract were determined to be pH 8.0-9.5 and 60 degrees C, respectively. Under the optimum conditions, 620 mg/l of albonoursin was obtained with a conversion ratio of 62% after 24 h incubation. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI: 10.1016/S1381-1177(98)00079-4

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  • Inhibition of plant transformation by phytolaccoside b from phytolacca americana callus

    Hiroshi Kanzaki, Toshihiko Kagemori, Yoko Yamachika, Teruhiko Nitoda, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   63 ( 9 )   1657 - 1659   1999.1

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    The newly established GUS expression bioassay on the callus extracts of 22 species of plants revealed that the methanol extract of Phytolacca americana callus had the most potent inhibitory activity against agrobacterial plant transformation. A triterpene glycoside phytolaccoside B was isolated from the extract as a genuine plant transformation inhibitor having neither antiagrobacterial nor phytotoxic activity. This compound is promising for use as a biochemical probe for studies on the plant transformation mechanism. © 1999, Taylor &amp
    Francis Group, LLC. All rights reserved.

    DOI: 10.1271/bbb.63.1657

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  • Inhibition of plant transformation by phytolaccoside b from phytolacca americana callus

    Hiroshi Kanzaki, Toshihiko Kagemori, Yoko Yamachika, Teruhiko Nitoda, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   63 ( 9 )   1657 - 1659   1999.1

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    The newly established GUS expression bioassay on the callus extracts of 22 species of plants revealed that the methanol extract of Phytolacca americana callus had the most potent inhibitory activity against agrobacterial plant transformation. A triterpene glycoside phytolaccoside B was isolated from the extract as a genuine plant transformation inhibitor having neither antiagrobacterial nor phytotoxic activity. This compound is promising for use as a biochemical probe for studies on the plant transformation mechanism. © 1999, Taylor &amp
    Francis Group, LLC. All rights reserved.

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  • Effective production of dehydro cyclic dipeptide albonoursin exhibiting pronuclear fusion inhibitory activity. I. Taxonomy and fermentation

    Hiroshi Kanzaki, Daisuke Imura, Reiko Sashida, Akio Kobayashi, Kazuyoshi Kawazu

    Journal of Antibiotics   52 ( 11 )   1017 - 1022   1999

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    Strain KO-23, an actinomycete producing albonoursin as well as streptopyrone, was identified as Streptomyces albulus by morphological and biochemical studies. Fermentation conditions for albonoursin, a dehydro cyclic dipeptide exhibiting a pronounced inhibitory activity toward pronuclear fusion of sea urchin eggs, were optimized. Under the optimum conditions, the actinomycete produced 16 mg/liter of albonoursin, 30 times higher than that in the original culture. The cells cultivated under these conditions highly express biosynthetic enzymes for albonoursin, and thus are available for biosynthetic studies of dehydro cyclic peptides.

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  • 病原体を接種して萎凋させたアカマツ実生からの病原体の再分離およびマツノザイセンチュウの分離系統間の病原性の差異.

    河津一儀, 金子昇, 平岡享子, 山下秀昭, 神崎浩

    岡山大学農学部学術報告   88   1 - 6   1999

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  • 新規の環状ジペプチド脱水素酵素とその活性測定法の確立.

    神崎浩, 赤澤和美, 井村大輔, 仁戸田照彦

    岡山大学農学部学術報告   1999

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  • Microbial hyrodolysis of cyclo(Gly-Leu)

    Kanzaki Hiroshi, Mizuta Naoki, Miyoshi Kazuyuki, Nitoda Teruhiko

    11   202 - 202   1999

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  • Calophyllum inophyllumの抗HIV活性成分inophyllum A,B,C,D,E,Pの液体クロマトグラフィー・による分析法の確立

    河津 一儀, 仁戸田 照彦, 神崎 浩

    岡山大学農学部学術報告   87   13 - 16   1998.2

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  • Novel bioactive oxazolomycin isomers produced by streptomyces albus JA3453

    Hiroshi Kanzaki, Ken-Ichi Wada, Teruhiko Nitoda, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   62 ( 3 )   438 - 442   1998

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    Two novel oxazolomycin isomers, oxazolomycins B (2) and C (3), were isolated from the fermentation broth of an oxazolomycin-producing strain, Streptomyces albus JA3453. Both compounds are geometrical isomers of oxazolomycin (1), the configurations of their triene moieties being (4′E, 6′E, 8′E) (2) and (4′Z, 6′E, 8′E) (3) while that of oxazolomycin (1) is (4′Z, 6′Z, 8′E). Compounds 2 and 3 exhibited potent inhibitory activity against crown gall formation with the same MIC (0.8 μg/disk) as oxazolomycin. Compounds 2 and 3 showed no antibacterial activity against Agrobacterium tumefaciens, in contrast to oxazolomycin which has specific anti-A. tumefaciens activity. © 1998, Taylor &amp
    Francis Group, LLC. All rights reserved.

    DOI: 10.1271/bbb.62.438

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  • Improved bioassay method for plant transformation inhibitors

    Hiroshi Kanzaki, Toshihiko Kagemori, Satomi Asano, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   62 ( 12 )   2328 - 2333   1998

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    A convenient and quantitative bioassay method for evaluating the efficiency of plant transformation by Agrobacterium tumefaciens is important to search plant transformation inhibitors, possible biochemical probes for study on its mechanism. Our previously reported method, in which the plant transformation had been detected by the expression of β-glucuronidase in transformed plants, was improved. The difference between the previous and the improved methods is the use of suspension-cultured cells of Ageratum conyzoides as the host plant instead of Nicotiana tabacum BY-2
    this alteration of the host enabled us to measure the β-glucuronidase activity in plant cells not only fluorometrically but also colorimetrically. The enzyme activity expressed in the cells of A. conyzoides was nearly 100 times higher than that of N. tabacum BY-2, and was enough for detection by colorimetric measurement. The method, therefore, is useful for a convenient determination of inhibitory activity against plant transformation. © 1998, Taylor &amp
    Francis Group, LLC. All rights reserved.

    DOI: 10.1271/bbb.62.2328

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  • Improved bioassay method for plant transformation inhibitors

    Hiroshi Kanzaki, Toshihiko Kagemori, Satomi Asano, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   62 ( 12 )   2328 - 2333   1998

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    A convenient and quantitative bioassay method for evaluating the efficiency of plant transformation by Agrobacterium tumefaciens is important to search plant transformation inhibitors, possible biochemical probes for study on its mechanism. Our previously reported method, in which the plant transformation had been detected by the expression of β-glucuronidase in transformed plants, was improved. The difference between the previous and the improved methods is the use of suspension-cultured cells of Ageratum conyzoides as the host plant instead of Nicotiana tabacum BY-2
    this alteration of the host enabled us to measure the β-glucuronidase activity in plant cells not only fluorometrically but also colorimetrically. The enzyme activity expressed in the cells of A. conyzoides was nearly 100 times higher than that of N. tabacum BY-2, and was enough for detection by colorimetric measurement. The method, therefore, is useful for a convenient determination of inhibitory activity against plant transformation. © 1998, Taylor &amp
    Francis Group, LLC. All rights reserved.

    DOI: 10.1271/bbb.62.2328

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  • Microbial hydrolysis of diketopiperazines: Different types of diketopiperazine-assimilating bacteria

    H Kanzaki, S Oda, A Kobayashi, K Kawazu

    JOURNAL OF FERMENTATION AND BIOENGINEERING   83 ( 4 )   386 - 388   1997

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    Arthrobacter sp. 1-3-1 and a coryneform rod bacterium, strain T-1-3-Y, which assimilate cyclo(Gly-Gly) and cyclo(Gly-L-Tyr) at high rates, respectively, were isolated from sell. Both bacteria exhibit diketopiperazine-hydrolyzing activity intracellularly in contrast to the extracellular cyclo(Gly-Gly) hydrolase reported previously. Arthrobacter sp. 1-3-1 shows a much broader substrate specificity than strain T-1-3-Y, indicating the diversity of diketopiperazine hydrolysis in nature.

    DOI: 10.1016/S0922-338X(97)80147-X

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  • Microbial hydrolysis of diketopiperazines: Different types of diketopiperazine-assimilating bacteria

    H Kanzaki, S Oda, A Kobayashi, K Kawazu

    JOURNAL OF FERMENTATION AND BIOENGINEERING   83 ( 4 )   386 - 388   1997

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    Arthrobacter sp. 1-3-1 and a coryneform rod bacterium, strain T-1-3-Y, which assimilate cyclo(Gly-Gly) and cyclo(Gly-L-Tyr) at high rates, respectively, were isolated from sell. Both bacteria exhibit diketopiperazine-hydrolyzing activity intracellularly in contrast to the extracellular cyclo(Gly-Gly) hydrolase reported previously. Arthrobacter sp. 1-3-1 shows a much broader substrate specificity than strain T-1-3-Y, indicating the diversity of diketopiperazine hydrolysis in nature.

    DOI: 10.1016/S0922-338X(97)80147-X

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  • Inhibitory effect of curromycins and their esters on plant transformation

    Hiroshi Kanzaki, Toshitsune Ichioka, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   60 ( 9 )   1535 - 1537   1996

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    Curromycins A and B, antibiotics specific to Agrobacterium tumefaciens, inhibited crown gall formation on potato tubers. Their esters showed potent inhibitory activity against crown gall formation without any antibacterial activity. The change in biological activity of curromycins by simple esterification was similar to our previous observation in oxazolomycin. Curromycin esters are possible chemical probes to study the mechanism for plant transformation. © 1996, Taylor &amp
    Francis Group, LLC. All rights reserved.

    DOI: 10.1271/bbb.60.1535

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  • Inhibitory effect of curromycins and their esters on plant transformation

    Hiroshi Kanzaki, Toshitsune Ichioka, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   60 ( 9 )   1535 - 1537   1996

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    Curromycins A and B, antibiotics specific to Agrobacterium tumefaciens, inhibited crown gall formation on potato tubers. Their esters showed potent inhibitory activity against crown gall formation without any antibacterial activity. The change in biological activity of curromycins by simple esterification was similar to our previous observation in oxazolomycin. Curromycin esters are possible chemical probes to study the mechanism for plant transformation. © 1996, Taylor &amp
    Francis Group, LLC. All rights reserved.

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  • An effective method of screening glucose-rich microbial culture filtrates for insect trehalase inhibitors

    Hiroshi Kanzaki, Takafumi Nuhama, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   59 ( 3 )   398 - 400   1995

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    Trehalase inhibitors are promising as a specific insect growth regulator. Microbial culture filtrates are a good source of bioactive compounds, but some of them retain a significant amount of glucose, a major component of the culture medium, which makes it difficult to detect trehalase inhibitors because of high blank values for the glucose determination. To find a variety of trehalase inhibitors in glucose-rich microbial culture filtrates, an effective method for removing glucose by enzymatic conversion prior to a routine trehalase-inhibitory assay was devised. Conversion with glucose oxidase and catalase could completely remove 10mg/ml of glucose, and a wide variety of microbial culture filtrates with high glucose content could then be subjected to the routine bioassay. This method can also be applied to screen inhibitors for other enzymes whose activity is assayed by the amount of glucose formed. © 1995 Taylor &amp
    Francis Group LLC.

    DOI: 10.1080/bbb.59.398

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  • An effective method of screening glucose-rich microbial culture filtrates for insect trehalase inhibitors

    Hiroshi Kanzaki, Takafumi Nuhama, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology and Biochemistry   59 ( 3 )   398 - 400   1995

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    Trehalase inhibitors are promising as a specific insect growth regulator. Microbial culture filtrates are a good source of bioactive compounds, but some of them retain a significant amount of glucose, a major component of the culture medium, which makes it difficult to detect trehalase inhibitors because of high blank values for the glucose determination. To find a variety of trehalase inhibitors in glucose-rich microbial culture filtrates, an effective method for removing glucose by enzymatic conversion prior to a routine trehalase-inhibitory assay was devised. Conversion with glucose oxidase and catalase could completely remove 10mg/ml of glucose, and a wide variety of microbial culture filtrates with high glucose content could then be subjected to the routine bioassay. This method can also be applied to screen inhibitors for other enzymes whose activity is assayed by the amount of glucose formed. © 1995 Taylor &amp
    Francis Group LLC.

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  • A novel bioassay method for inhibitors against plant transformation

    Kanzaki Hiroshi, Toyoshima Hiroto, Kobayashi Akio, Kawazu Kazuyoshi

    Bioscience, Biotechnology and Biochemistry   58 ( 3 )   531 - 534   1994.3

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    A rapid and quantitative bioassay method for inhibitors of plant transformation by Agrobacterium tumefaciens with an intron-GUS binary vector was devised as an alternative method to the time-consuming potato tuber disk assay. Among several host plants tested, tobacco BY-2 suspension cells were the most preferable for this GUS expression assay. Furthermore, as the phytotoxicity of the test material, if any, is apparent from the growth inhibition of BY-2 suspension cells, this method can exclude phytotoxic com pounds.

    DOI: 10.1271/bbb.58.531

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  • A Novel Bioassay Method for Inhibitors of Plant Transformation

    Kanzaki Hiroshi, Toyoshima Hiroto, Kobayashi Akio, Kawazu Kazuyoshi

    Bioscience, biotechnology, and biochemistry   58 ( 3 )   531 - 534   1994.3

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    A rapid and quantitative bioassay method for inhibitors of plant transformation by Agrobacterium tumefaciens with an intron-GUS binary vector was devised as an alternative method to the time-consuming potato tuber disk assay. Among several host plants tested, tobacco BY-2 suspension cells were the most preferable for this GUS expression assay. Furthermore, as the phytotoxicity of the test material, if any, is apparent from the growth inhibition of BY-2 suspension cells, this method can exclude phytotoxic com pounds.

    DOI: 10.1271/bbb.58.531

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  • 47 THE ROLE OF PEROXIDASE IN THE CHEMICAL DEFENCE OF PLANTS : A NOVEL PEROXIDASE INHIBITOR AND A NEW TYPE OF BIOACTIVE PHENOLICS PRODUCED BY PEROXIDASE

    Kobayashi A, Koguchi Y, Kanzaki H, Kajiyama S, Kawazu K

    天然有機化合物討論会講演要旨集   ( 35 )   361 - 368   1993.9

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    Plants induce varied resistance-response toward microbial challenge. In order to establish an infection, pathogenic fungi must break such defense barriers as lignin, phytoalexins, and hydrolytic enzymes. At the early stage of infection, POD (peroxidase) activity is enhanced dramatically. POD and lignin accumulates at the infected site and then protects from further fungal invasion. The suppression of lignin deposition at the site might be of advantage to susceptible pathogenic fungi. We, therefore, conducted a comprehensive screening to obtain POD inhibitors from fungal cultures and a potent radical scavenger named ST-1 was isolated from a fungus, and its chemical structure was elucidated to be 1,3-dihydro-4,5,6-trihydroxy-1,3-dimethoxy-7-methylisobenzofuran by a combination of chemical modification and spectroscopic techniques. ST-1 effectively scavenged oxygen radicals and also suppressed the phytoalexin accumulation in kidney bean. These results suggested that such a mechanism can be enumerated as one of significant defense systems intervening plants and microbes. On the other hand, the POD, activated upon pathogenic challenge, could convert common plant phenolics into bioactive compounds which interfered with pathogen and insect attacks. This idea prompted us to examine the possibility. Guaiacol and other phenols were chosen as model substrates for the enzyme reaction system, and several products with novel structures were confirmed to have unique biological activities. This paper deals with the chemical structures of a novel radical scavenger (ST-1) and the POD products with anti-microbial activities and phytotoxicities.

    DOI: 10.24496/tennenyuki.35.0_361

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  • Bisanthraquinones, Inhibitors of Plant Transformation

    Hiroshi Kanzaki, Goro Kawabata, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology, and Biochemistry   57 ( 12 )   2104 - 2106   1993

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    A potato tuber disk assay of culture broths which had been confirmed to show neither antibacterial nor phytotoxic activity resulted in the discovery of a novel type of crown gall formation inhibitors, julimycin B-II and julichrome Q1.3. These two bisanthraquinones, products of Streptomyces sp. TM-71, inhibited crown gall formation on potato tuber disks at a minimum inhibitory dose of 1.6μg/disk without affecting the growth of Agrobacterium tumefaciens or the germination of alfalfa seeds. © 1993, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb.57.2104

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  • Bisanthraquinones, Inhibitors of Plant Transformation

    Hiroshi Kanzaki, Goro Kawabata, Akio Kobayashi, Kazuyoshi Kawazu

    Bioscience, Biotechnology, and Biochemistry   57 ( 12 )   2104 - 2106   1993

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    A potato tuber disk assay of culture broths which had been confirmed to show neither antibacterial nor phytotoxic activity resulted in the discovery of a novel type of crown gall formation inhibitors, julimycin B-II and julichrome Q1.3. These two bisanthraquinones, products of Streptomyces sp. TM-71, inhibited crown gall formation on potato tuber disks at a minimum inhibitory dose of 1.6μg/disk without affecting the growth of Agrobacterium tumefaciens or the germination of alfalfa seeds. © 1993, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb.57.2104

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  • Oxazolomycin Esters, Specific Inhibitors of Plant Transformation

    Kazuyoshi Kawazu, Goro Kawabata, Satoru Kawai, Akio Kobayashi, Hiroshi Kanzaki

    Bioscience, Biotechnology and Biochemistry   56 ( 9 )   1382 - 1385   1992

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    Oxazolomycin diacetate, dipropionate, monobutyrate and dibutyrate were derived from oxazolomycin, a product of Streptomyces sp. KBFP-2025. These esters were potent inhibitors of crown gall formation on potato tuber disks upon infection with Agrobacterium tumefaciens. They showed neither antibacterial nor phytotoxic activity, whereas oxazolomycin showed both antibacterial and phytotoxic activities. Further, they had no inhibitory activity against A. tumefaciens on the potato tuber disk. The inhibitory activity of these esters against crown gall formation seems to be due to specific inhibition of the transformation of plants with A. tumefaciens. © 1992, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb.56.1382

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  • Oxazolomycin Esters, Specific Inhibitors of Plant Transformation

    Kazuyoshi Kawazu, Goro Kawabata, Satoru Kawai, Akio Kobayashi, Hiroshi Kanzaki

    Bioscience, Biotechnology and Biochemistry   56 ( 9 )   1382 - 1385   1992

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    Oxazolomycin diacetate, dipropionate, monobutyrate and dibutyrate were derived from oxazolomycin, a product of Streptomyces sp. KBFP-2025. These esters were potent inhibitors of crown gall formation on potato tuber disks upon infection with Agrobacterium tumefaciens. They showed neither antibacterial nor phytotoxic activity, whereas oxazolomycin showed both antibacterial and phytotoxic activities. Further, they had no inhibitory activity against A. tumefaciens on the potato tuber disk. The inhibitory activity of these esters against crown gall formation seems to be due to specific inhibition of the transformation of plants with A. tumefaciens. © 1992, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb.56.1382

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  • Preparation and identification of the alfalfa elicitor from brown algae laminaran.

    KOBAYASHI Akio, TAI Akihiro, KANZAKI Hiroshi, KAWAZU Kazuyoshi

    Chemical Regulation of Plants   26 ( 2 )   222 - 222   1991.12

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  • Production of Benzoylformic Acid from Phenylglycine by Saccharomycopsis lipolytica

    Hiroshi Kanzaki, Atsuhiko Isobe, Yoshikazu Izumi, Hideaki Yamada

    Agricultural and Biological Chemistry   54 ( 8 )   2101 - 2105   1990

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    Microbial production of benzoylformic acid (BF), which can be used as a substrate of enzymatic synthesis of (R)-(—)-mandelic acid, was investigated. Among 145 strains of yeasts and actinomycetes, Saccharomycopsis lipolytica (IAM 4964) was the best producer of BF from DL-phenylglycine (DL-PG). Culture conditions for BF production by the organism were optimized. When 0.2 % fructose as a carbon source and 0.7% Bacto-tryptone as a nitrogen source were used in the presence of 4 % dl-PG, 14.5 mg/ml of BF was produced (about 37 % molar yield) in 4 days of cultivation. BF was synthesized from the L-form of PG, but not from the D-form. The BF was isolated from culture broth in a crystalline form and physicochemically identified. © 1990, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb1961.54.2101

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  • Production of Benzoylformic Acid from Phenylglycine by Saccharomycopsis lipolytica

    Hiroshi Kanzaki, Atsuhiko Isobe, Yoshikazu Izumi, Hideaki Yamada

    Agricultural and Biological Chemistry   54 ( 8 )   2101 - 2105   1990

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    Microbial production of benzoylformic acid (BF), which can be used as a substrate of enzymatic synthesis of (R)-(—)-mandelic acid, was investigated. Among 145 strains of yeasts and actinomycetes, Saccharomycopsis lipolytica (IAM 4964) was the best producer of BF from DL-phenylglycine (DL-PG). Culture conditions for BF production by the organism were optimized. When 0.2 % fructose as a carbon source and 0.7% Bacto-tryptone as a nitrogen source were used in the presence of 4 % dl-PG, 14.5 mg/ml of BF was produced (about 37 % molar yield) in 4 days of cultivation. BF was synthesized from the L-form of PG, but not from the D-form. The BF was isolated from culture broth in a crystalline form and physicochemically identified. © 1990, Japan Society for Bioscience, Biotechnology, and Agrochemistry. All rights reserved.

    DOI: 10.1271/bbb1961.54.2101

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  • Characterization of crystalline formate dehydrogenase from Candida methanolica

    Yoshikazu IZUMI, Hiroshi KANZAKI, Shigeru MORITA, Hideaki FUTAZUKA, Hideaki YAMADA

    European Journal of Biochemistry   182 ( 2 )   333 - 341   1989

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    The crystalline formate dehydrogenase from Candida methanolica, which showed the highest specific activity (7.52 U/mg) so far reported, was characterized in detail. The enzyme is a dimer composed of identifical subunits, each containing one SH group related to the catalytic activity. The molecular mass of the enzyme is about 82–86 kDa. The Km values were found to be 3.0 mM for formate and 0.11 mM for NAD+. Even if the enzyme was incubated at pH 6.5–9.5 or at 55°C, the activity remained at 100%. Hg2+, Ni2+, NaCN, NaN3 and p‐chloromercuribenzoate strongly inhibited the enzyme activity, while the enzyme showed relatively high resistance to various chelating agents. The amino acid composition and some other physicochemical properties of the enzyme were studied. Immunological studies revealed that formate dehydrogenases of methanol‐utilizing yeasts immunologically more or less resemble each other, but differ from those of methanol‐utilizing bacteria. Furthermore, yeast formate dehydrogenases can be immunologically classified into three types: (a) the Candida type, (b) the Torulopsis/Hansenula/Pichia type and (c) the formaldehyde‐resistant yeast type. For simple and large‐scale preparation of the enzyme for practical use, treatment of cells of C. methanolica with the commercial cationic detergent, ‘Benzalkonium’ cation, is useful: the total and specific activities of the enzyme are 1.17‐fold and 3.10‐fold higher than those of the crude cell‐free extract, respectively. Copyright © 1989, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1432-1033.1989.tb14835.x

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  • Characterization of crystalline formate dehydrogenase from Candida methanolica

    Yoshikazu IZUMI, Hiroshi KANZAKI, Shigeru MORITA, Hideaki FUTAZUKA, Hideaki YAMADA

    European Journal of Biochemistry   182 ( 2 )   333 - 341   1989

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    The crystalline formate dehydrogenase from Candida methanolica, which showed the highest specific activity (7.52 U/mg) so far reported, was characterized in detail. The enzyme is a dimer composed of identifical subunits, each containing one SH group related to the catalytic activity. The molecular mass of the enzyme is about 82–86 kDa. The Km values were found to be 3.0 mM for formate and 0.11 mM for NAD+. Even if the enzyme was incubated at pH 6.5–9.5 or at 55°C, the activity remained at 100%. Hg2+, Ni2+, NaCN, NaN3 and p‐chloromercuribenzoate strongly inhibited the enzyme activity, while the enzyme showed relatively high resistance to various chelating agents. The amino acid composition and some other physicochemical properties of the enzyme were studied. Immunological studies revealed that formate dehydrogenases of methanol‐utilizing yeasts immunologically more or less resemble each other, but differ from those of methanol‐utilizing bacteria. Furthermore, yeast formate dehydrogenases can be immunologically classified into three types: (a) the Candida type, (b) the Torulopsis/Hansenula/Pichia type and (c) the formaldehyde‐resistant yeast type. For simple and large‐scale preparation of the enzyme for practical use, treatment of cells of C. methanolica with the commercial cationic detergent, ‘Benzalkonium’ cation, is useful: the total and specific activities of the enzyme are 1.17‐fold and 3.10‐fold higher than those of the crude cell‐free extract, respectively. Copyright © 1989, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1432-1033.1989.tb14835.x

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  • Purification and characterization of cystathionine γ‐synthase type II from Bacillus sphaericus

    Hiroshi KANZAKI, Michihiko KOBAYASHI, Toru NAGASAWA, Hideaki YAMADA

    European Journal of Biochemistry   163 ( 1 )   105 - 112   1987

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    Cystathionine γ‐synthase type II, which catalyzes l‐cystathionine synthesis from O‐acetyl‐l‐homoserine and l‐cysteine was purified from Bacillus sphaericus (IFO 3536) in seven steps. The purified enzyme appeared to be homogeneous by the results of polyacrylamide electrophoresis and ampholyte electrofocusing. The enzyme is a typical pyridoxal‐P dependent enzyme, has a molecular mass of 165 kDa and consists of four subunits identical in molecular mass. The enzyme catalyzed the γ‐replacement reaction and the elimination reaction was hardly detected even when a large amount of enzyme was added. In the replacement reaction, O‐acetyl‐l‐homoserine and the following thiol compounds: L and d‐cysteine, L and d‐homocysteine, sodium sulfide, various alkyl and aryl mercaptans, acted as the most suitable substrate to produce l‐cystathionine and the corresponding S‐substituted l‐homocysteine derivatives. Copyright © 1987, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1432-1033.1987.tb10742.x

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  • Purification and characterization of cystathionine γ‐synthase type II from Bacillus sphaericus

    Hiroshi KANZAKI, Michihiko KOBAYASHI, Toru NAGASAWA, Hideaki YAMADA

    European Journal of Biochemistry   163 ( 1 )   105 - 112   1987

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    Cystathionine γ‐synthase type II, which catalyzes l‐cystathionine synthesis from O‐acetyl‐l‐homoserine and l‐cysteine was purified from Bacillus sphaericus (IFO 3536) in seven steps. The purified enzyme appeared to be homogeneous by the results of polyacrylamide electrophoresis and ampholyte electrofocusing. The enzyme is a typical pyridoxal‐P dependent enzyme, has a molecular mass of 165 kDa and consists of four subunits identical in molecular mass. The enzyme catalyzed the γ‐replacement reaction and the elimination reaction was hardly detected even when a large amount of enzyme was added. In the replacement reaction, O‐acetyl‐l‐homoserine and the following thiol compounds: L and d‐cysteine, L and d‐homocysteine, sodium sulfide, various alkyl and aryl mercaptans, acted as the most suitable substrate to produce l‐cystathionine and the corresponding S‐substituted l‐homocysteine derivatives. Copyright © 1987, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1432-1033.1987.tb10742.x

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  • Insight into the active site of Streptomyces cystathionine γ-lyase based on the results of studies on its substrate specificity

    KANZAKI H.

    Biochim. Biophys. Acta   913   45 - 50   1987

  • Highly efficient production of l-cystathionine from O-succinyl-l-homoserine and l-cysteine by Streptomyces cystathionine γ-lyase

    Hiroshi Kanzaki, Toru Nagasawa, Hideaki Yamada

    Applied Microbiology and Biotechnology   25 ( 2 )   97 - 100   1986

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    Utilizing the γ-replacement reaction of Streptomyces cystathionine γ-lyase (EC 4.4.1.1.), an efficient production method for l-cystathionine has been established. Under optimal conditions, 50 mM l-cystathionine was synthesized from 50 mM O-succinyl-l-homoserine and 50 mM l-cysteine, added in four stages to the reaction mixture, with a substrate conversion rate of 100%. This productivity (11 gl-1 of reaction mixture) is about 3.5 times higher than that with l-homoserine and l-cysteine as substrates. © 1986 Springer-Verlag.

    DOI: 10.1007/BF00250516

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Presentations

  • Microbial Conversion of an Olive Leaf Extract Constituent to Create a Novel Antioxidative Compound B-olivol® as a Cosmetic Material Invited

    International Conference on Contemporary Science and Clinical Pharmacy (ICCSCP) 2023  2023.10.31 

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    Event date: 2023.10.30 - 2023.10.31

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  • Biocatalyst-mediated synthesis of bioactive ‘Unnatural’ natural products Invited

    Hiroshi Kanzaki

    The 23rd Symposium of the Society of Biocatalysis Japan  2023.9.29 

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    Event date: 2023.9.28 - 2023.9.29

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  • Inhibitory sensitivity of β-N-acetylglucosaminidase from Enterococcus sp. MK26 to N,N,N-trimethyl-β-D-glucosaminium-containing compounds

    K. Morioka, H. Ono, M. Koide, S. Suganuma, T. Nitoda, H. Kanzaki

    The 23rd Symposium of the Society of Biocatalysis Japan  2023.9.28 

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    Event date: 2023.9.28 - 2023.9.29

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  • Distribution of mammalian gut microbiota capable of converting C-glycoside mangiferin into its aglycone norathyriol

    Y. Uga, U. Hasanah, H. Kayaki, H. Kawakami, Y. Nishitani, H. Kuwahara, T. Nitoda, H. Kanzaki

    The 23rd Symposium of the Society of Biocatalysis Japan  2023.9.28 

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    Event date: 2023.9.28 - 2023.9.29

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  • Pochonia suchlasporia が生産するasteltoxin 類のMS/MS 分析を用いた探索

    加藤陽輝, 神崎 浩, 仁戸田照彦

    日本農芸化学会2023 年度中四国・西日本支部合同大会 (中四国支部第66 回・西日本支部第347 回講演会)  2023.9.22 

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    Event date: 2023.9.21 - 2023.9.22

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  • Cyclo(Leu-Phe) oxidase が触媒する環状ジペプチド脱水素反応の反応初期における人工電子受容体添加の影響の定量的評価

    猪口慧悟, 坂口幸士朗, 仁戸田照彦, 神崎浩

    日本農芸化学会2023 年度中四国・西日本支部合同大会 (中四国支部第66 回・西日本支部第347 回講演会)  2023.9.22 

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    Event date: 2023.9.21 - 2023.9.22

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  • 微生物変換で天然に存在しない 生物活性天然有機化合物を創生する Invited

    神崎 浩

    第40回農薬環境科学研究会 グローバルにアプローチするグリーンテクノロジー  2023.9.14 

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    Event date: 2023.9.14 - 2023.9.15

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  • Microbial conversion of C-glycoside mangiferin into its aglycone norathyriol by mammalian intestinal bacteria

    Y. Uga, U. Hasanah, H. Kayaki, H. Kawakami, Y. Nishitani, H. Kuwahara, T. Nitoda, H. Kanzaki

    2023.9.8 

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    Event date: 2023.9.8 - 2023.9.9

    Language:Japanese   Presentation type:Poster presentation  

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  • 麹菌固体培養技術を活かした植物資源の高機能化

    神崎 浩

    国際発酵・醸造食品産業展2023 発酵アカデミックフォーラム  2023.8.4 

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    Event date: 2023.8.2 - 2023.8.4

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Bioconversion of olive leaf constituents by Aspergillus solid state fermentation using an air flow type rotary solid state fermenter for laboratory use

    ○Nanami TATSUMI, Atsuko HASHIMOTO, Hinano OKUKAWA, Tsuyoshi MIYAKE, Kazunari ITO, Yuka TANINO, Hayato TAKEUCHI, Hideyuki YAMASHITA, Mami UCHIDA, Shouhei MIKI, Yasuhiro YOSHIDA, Megumi JYO, Keiichi KIKUCHI, Natsuki FUKANO, Teruhiko NITODA, Hiroshi KANZAKI

    2023.3.15 

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    Event date: 2023.3.14 - 2023.3.17

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Bioconversion of skin and seed constituents by Aspergillus solid state fermentation of wine pomace

    Ayaka FUJITA, Hinano OKUKAWA, Atsuko HASHIMOTO, Tsuyoshi MIYAKE, Kazunari ITO, Yuka TANINO, Hideyuki YAMASHITA, Takuro NAKAGAWA, Koji HIRANO, Teruhiko NITODA, Hiroshi KANZAKI

    2023.3.15 

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    Event date: 2023.3.14 - 2023.3.17

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Structure Analysis of Antifungal Sesterterpene ophiobilin K and Its Analogs Produced by Aspergillus ustus

    Nguyen T.K. Ngoc, Natthapat Sohsomboon, Terumi Ichimiya, Teruhiko Nitoda, Hiroshi Kanzaki

    2022.11.18 

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    Event date: 2022.11.17 - 2022.11.19

    Language:English   Presentation type:Oral presentation (general)  

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  • Comparison of Thai and Japanese Fermented Milk Produced by Potential Probiotic Lactic Acid Bacteria Isolated in Thailand

    Jirayu Jitpakdee, Hiroshi Kanzaki, Teruhiko Nitoda, Duangporn Kantachote

    The international conference on Sustainable Agriculture and Environment (SAE)  2022.11.18 

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    Event date: 2022.11.17 - 2022.11.19

    Language:English   Presentation type:Oral presentation (general)  

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  • Microbial conversion process of an olive leaf extract to produce a novel antioxidative compound B-olivol® as a cosmetic Invited

    Hiroshi Kanzaki, Hisao Moriya, Megumi Jyo, Keiichi Kikuchi, Yasuhiro Yoshida, Teruhiko Nitoda

    Active Enzyme Molecule 2022  2022.9.30 

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    Event date: 2022.9.30 - 2022.10.1

    Language:English   Presentation type:Oral presentation (invited, special)  

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  • ウマ腸内細菌叢によるC-配糖体mangiferinからそのアグリコンnorathyriolへの変換

    宇賀悠人, 川上秀昭, 西谷洋輔, 桑原浩誠, 仁戸田照彦, 神崎 浩

    日本農芸化学会 2022年度中四国支部大会(第63回講演会)  2022.9.22 

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    Event date: 2022.9.21 - 2022.9.22

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Enterococcus sp. MK26株由来β-N-acetylglucosaminidaseの N,N,N-trimethyl-β-D-glucosaminium化合物に対する阻害感受性

    森岡京平, 小野はるか, 小出麻奈, 菅沼笙子, 仁戸田照彦, 神崎浩

    おかやまバイオアクティブ研究会 第61回シンポジウム  2022.9.20 

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    Event date: 2022.9.20

    Language:Japanese   Presentation type:Poster presentation  

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  • 微生物発酵でおもろいもんを作る Invited

    神崎 浩

    超異分野学会 大阪大会2022 発酵リモデリングによる物質の価値創造〜発酵をアップデートする〜  2022.8.27 

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    Event date: 2022.8.27

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

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  • バイオサーファクタント・サーファクチンの自己集合特性への対カチオン種の違いによる影響

    柳澤恵広, 平 敏彰, 仁戸田照彦, 神崎 浩

    日本農芸化学会中四国支部 第62回支部講演会(例会)  2022.6.4 

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    Event date: 2022.6.4

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • ワインパミスの麹菌固体培養における熱水処理の影響

    奥川 日菜乃, 橋本 敦子, 三宅 剛史, 伊藤 一成, 谷野 有佳, 山下 秀行, 中川 拓郎, 平野 幸司, 仁戸田 照彦, 神崎 浩

    日本農芸化学会2022年度京都大会  2022.3.16 

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    Event date: 2022.3.15 - 2022.3.18

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • オリーブ葉の麹菌固体培養における蒸煮処理の影響

    橋本 敦子, 奥川 日菜乃, 三宅 剛史, 伊藤 一成, 谷野 有佳, 山下 秀行, 内田 真美, 三木 翔平, 吉田 靖弘, 徐 恵美, 菊地 敬一, 仁戸田 照彦, 神崎 浩

    日本農芸化学会2022年度京都大会  2022.3.16 

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    Event date: 2022.3.15 - 2022.3.18

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  • MS/MS分析による効率的な新規asteltoxin類探索法の検討

    三谷尚大, 神崎浩, 仁戸田照彦

    日本農薬学会第47回大会(岡山)  2022.3.8 

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    Event date: 2022.3.7 - 2022.3.9

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • Microbial conversion of constituents of Eucommia ulmoides leaves into useful compounds

    仁戸田照彦, 神崎浩

    日本杜仲研究会定期大会講演要旨集  2021 

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  • Penicillium aurantiogriseumが触媒するAspergillus ustus代謝物の新規フロイソキノリンアルカロイドへの変換反応

    山田光代, 齊藤太樹, 小川凌太, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2021 

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  • High functionalization of Japanese wine pomace constituents by Aspergillus solid-state fermentation

    奥川日菜乃, 橋本敦子, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 中川拓郎, 平野幸司, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2021 

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  • 麹菌固体培養による杜仲葉二次代謝産物の微生物変換

    村上響, 奥川日菜乃, 山下秀行, 三木翔平, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2021 

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  • High functionalization of olive leaf constituents by Aspergillus solid state fermentation

    橋本敦子, 奥川日菜乃, 三宅剛史, 伊藤一成, 谷野有佳, 山下秀行, 内田真美, 三木翔平, 吉田靖弘, 徐恵美, 菊地敬一, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2021 

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  • Investigation of the sensitivity of β-N-acetylglucosaminidase from p-nitrophenyl N-acetyl-β-D-glucosaminide assimilating bacteria to N,N,N-trimethylglucosaminium containing compounds

    小出麻奈, 菅沼笙子, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2020 

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  • Pochonia suchlasporiaの固体培養により生産される糸状菌胞子発芽阻害物質(第2報)

    片岡智範, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2020 

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  • エノキタケを栽培した針葉樹菌床の水抽出物が示す脂肪蓄積抑制効果

    畑本梨衣, 永谷直哉, 仁戸田照彦, 畑生俊光, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2020 

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  • Synthesis of pNP-TMG via electrochemical glycosylation

    桑名春希, 矢野君晟, 野上敏材, 仁戸田照彦, 神崎浩

    日本化学会春季年会講演予稿集(CD-ROM)  2020 

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  • A novel furo-isoqunoline alkaloid formed from Aspergillus ustus metabolites by Penicillium aurantiogriseum catalyst

    山田光代, 齊藤太樹, 小川凌太, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2020 

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  • Aspergillus ustusとPenicillium aurantiogriseumの組み合わせ培養により生産される新規フロイソキノリンアルカロイドの絶対立体配置の決定

    山田光代, 齊藤太樹, 小川凌太, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2019 

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  • 過酸化水素定量を利用したCyclo(Leu-Phe)oxidaseの活性測定法の確立と基質特異性検討

    小西健太, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2019 

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  • マウス腸内細菌叢によるC-配糖体mangiferinからそのアグリコンnorathyriolへの変換

    三木香澄, 川上秀昭, 西谷洋輔, 桑原浩誠, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2019 

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  • オリーブ葉二次代謝産物oleuropein aglyconのエピマーとその微生物還元生成物の構造解析

    栗原小蒔, 古賀まり子, 長谷井拓真, 菊地敬一, 徐恵美, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2019 

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  • Aspergillus ustusとPenicillium aurantiogriseumの組み合わせ培養により生産される新規フロイソキノリンアルカロイド

    山田光代, 齊藤太樹, 小川凌太, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2019 

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  • Pochonia suchlasporiaの固体培養により生産される糸状菌胞子発芽阻害物質

    片岡智範, 神崎浩, 仁戸田照彦

    日本農芸化学会大会講演要旨集(Web)  2019 

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  • Penicillium multicolorが生産する3種のp-nitrophenyl β-D-glucopyranoside分解酵素が触媒するL-アスコルビン酸への糖転移反応における基質特異性の比較研究

    小出麻奈, 菅沼笙子, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2018 

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  • オリーブ葉二次代謝産物3,4-dihydroxyphenylethanol-elenolic acidの還元を触媒するSaccharomyces cerevisiae由来酵素の探索と同定

    松川加奈, 巻尾沙織, 仁戸田照彦, 守屋央郎, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2018 

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  • β-N-Acetylglucosaminidase阻害物質TMG-chitotriomycin類縁体のLC-MSによる安定性および構造の検討

    田淵勇, 淺尾昂平, 水田雄大, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2018 

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  • Penicillium multicolorが生産する3種のp-nitrophenyl β-D-glucopyranosideの分解酵素の基質特異性に関する比較研究

    小出麻奈, 菅沼笙子, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2018 

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  • Saccharomyces cerevisiae由来alcohol dehydrogenase6は3,4-dihydroxyphenylethanol-elenolic acidの還元を触媒する

    松川加奈, 巻尾沙織, 仁戸田照彦, 守屋央郎, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2018 

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  • 糖供与体選択性の異なるβ-グルコシダーゼが触媒するL-アスコルビン酸への糖転移反応の速度論的研究

    菅沼笙子, 寺坂美紀, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2017 

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  • Cyclo(Leu-Phe)酸化酵素が有する広い基質特異性を利用したMet含有環状ジペプチドの変換反応

    小西健太, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2017 

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  • Aspergillus ustusとPenicillium aurantiogriseumの組み合わせ培養により生産される新規イソキノリンアルカロイドの生成経路

    小川凌太, 齊藤太樹, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2017 

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  • β-N-Acetylglucosaminidase阻害物質pochonicineのアセチル化によるNOE相関への影響

    江本啓輔, 奥田徹, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2016 

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  • オリーブ葉二次代謝産物oleuropein aglycon及びその微生物変換により生成するreduced oleuropein aglyconの詳細構造解析

    古賀まり子, 栗原小蒔, 徐恵美, 菊地敬一, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2016 

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  • L-アスコルビン酸への糖転移反応における基質特異性および位置特異性の異なるβ-グルコシダーゼの速度論的研究

    菅沼笙子, 寺坂美紀, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2016 

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  • オリーブ葉二次代謝産物oleuropein aglyconをreduced oleuropein aglyconへと変換するSaccharomyces cerevisiae由来酵素の探索

    巻尾沙織, 森吉弘, 仁戸田照彦, 守屋央朗, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2016 

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  • β-N-Acetylglucosaminidase阻害物質pochonicineおよびその類縁体の構造活性相関:6位と7位の水酸基の寄与

    虫明優一, 土田彩, 奥田徹, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2016 

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  • 食用キノコ処理木質バイオマスの家畜飼料素材としての利用

    永谷直哉, 小島豪, 柏野泰章, 柏野泰章, 長尾伸一郎, 時本景亮, 西野直樹, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集(Web)  2015 

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  • アスコルビン酸の位置特異的な配糖化を触媒するAspergillus niger由来β-グルコシダーゼを用いたアスコルビン酸6βグルコシドの合成

    寺坂美紀, 早川有美, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集  2012 

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  • 糸状菌Paecilomyces sp.F30株の生産するβ-N-Acetylglucosaminidase阻害剤

    戎義子, 奥田徹, 神崎浩, 仁戸田照彦

    生体触媒化学シンポジウム講演要旨集  2012 

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  • アスコルビン酸の位置特異的な配糖化を触媒するAspergillus niger由来β-グルコシダーゼを用いたアスコルビン酸6βグルコシドの合成

    寺坂美紀, 早川有美, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2012 

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  • ヒノキ木粉の生分解性を向上させる白色腐朽菌の探索

    小島豪, 光永均, 時本景亮, 西野直樹, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集  2012 

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  • β-N-Acetylglucosaminidase阻害剤pochonicineのGC-MS分析及び新規類縁体の確認

    土田彩, 安井あゆみ, 奥田徹, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集  2012 

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  • Cyclo(Leu-Phe)oxidase変換反応を受ける糸状菌由来二次代謝産物の検出

    高橋佳子, 福田優, 仁戸田照彦, 神崎浩

    日本農芸化学会大会講演要旨集(Web)  2012 

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  • A202 MS-MS fragmentation-guided search of TMG-chitotriomycin analogs and its structure-activity relationship

    Kanzaki Hiroshi, Usuki Hirokazu, Nitoda Teruhiko, Hatanaka Tadashi

    2011.2.25  Pesticide Science Society of Japan

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  • A203 Search for the analogs of β-N-acetylglucosaminidase inhibitor pochonicine produced by a fungal strain and improvement of their culture productivity

    Nitoda Teruhiko, Yamada Asami, Toyo-oka Miho, Usuki Hirokazu, Okuda Toru, Kanzaki Hiroshi

    2011.2.25  Pesticide Science Society of Japan

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    Language:Japanese  

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  • β-N-Acetylglucosaminidase阻害物質pochonicineとその類縁体の培養生産

    山田あさ美, 豊岡実穂, 臼木博一, 臼木博一, 奥田徹, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集  2011 

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  • Oleuropein aglyconの微生物変換で得られる還元化合物の詳細な抗酸化活性評価

    高津綾香, 秋久恵里佳, 吉田靖弘, 徐恵美, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2011 

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  • 精製CFL oxidaseの凝集性確認と可溶化の試み

    相澤ゆかり, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2011 

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  • β-N-Acetylglucosaminidase阻害物質TMG-chitotriomycinの阻害特性

    塩田博人, 行本千恵, 臼木博一, 臼木博一, 神崎浩, 仁戸田照彦

    日本農芸化学会大会講演要旨集  2011 

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  • 鰹節生産糸状菌が生産する抗酸化活性物質

    高橋佳子, 高津綾香, 福田優, 土居幹冶, 小山大, 仁戸田照彦, 神崎浩

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集  2011 

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  • 膜面液体培養による生理活性環状ジペプチド類の生産

    今宮亮, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集  2011 

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  • 発酵食品微生物が生産する抗酸化活性環状ジペプチドの酵素変換手法を用いる検出

    神崎浩, 高津綾香, 福田優, CHAO Man, 仁戸田照彦

    日本放線菌学会大会講演要旨集  2011 

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  • β-N-Acetylglucosaminidase阻害物質TMG-chitotriomycinの阻害選択性

    塩田博人, 臼木博一, 臼木博一, 神崎浩, 仁戸田照彦

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集  2011 

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  • 微生物変換で得られるoleuropein aglyconの還元型化合物およびその関連化合物の抗酸化活性評価

    高津綾香, 秋久恵里佳, 吉田靖弘, 徐恵美, 仁戸田照彦, 神崎浩

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集  2011 

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  • 放線菌Streptomyces anulatusにより生産されるβ-N-acetylglucosaminidase阻害剤の阻害選択性

    塩田博人, 臼木博一, 臼木博一, 神崎浩, 仁戸田照彦

    日本放線菌学会大会講演要旨集  2011 

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  • β-N-アセチルグルコサミニダーゼ阻害物質生産菌Streptomyces anulatusによるキチナーゼ阻害物質の生産

    塩田博人, 市川美里, 臼木博一, 神崎浩, 仁戸田照彦

    生体触媒化学シンポジウム講演要旨集  2010 

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  • β-N-アセチルグルコサミニダーゼ阻害物質生産菌Streptomyces anulatusにより生産されるキチナーゼ阻害物質の予備的性状解析

    塩田博人, 市川美里, 神崎浩, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集  2010 

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  • アスコルビン酸への糖転移反応を触媒しアスコルビン酸6βグルコシドを生成するAspergillus niger由来酵素(第2報)

    早川有美, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2010 

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  • アスコルビン酸の6位水酸基を特異的にβ-グルコシル化するAspergillus niger由来酵素(第2報)-糖転移活性の詳細検討-

    早川有美, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集  2010 

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  • 乳酸菌が生産する環状ジペプチド類の酵素法による検出

    張曼, ARUNRATTIYAKORN Panarat, 烏力吉徳力根, 宮本拓, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部講演会講演要旨集  2010 

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  • アスコルビン酸の6位水酸基を特異的にβ-グルコシル化するAspergillus niger由来酵素

    早川有美, 水津佳子, 仁戸田照彦, 神崎浩

    日本農芸化学会関西支部講演会講演要旨集  2009 

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  • アスコルビン酸への糖転移反応を触媒しアスコルビン酸6βグルコシドを生成するAspergillus niger由来酵素

    早川有美, 水津佳子, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2009 

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  • Streptomyces albulus KO23株由来組換えcyclo(Leu-Phe)oxidaseの精製と諸性質

    今村六美, 畑中唯史, 臼木博一, 仁戸田照彦, 神崎浩

    生体触媒化学シンポジウム講演要旨集  2009 

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  • β-N-アセチルグルコサミニダーゼ阻害剤MT-1の合成

    佐藤瑞穂, 豊岡実穂, 臼木博一, 仁戸田照彦, 神崎浩, 中島修平, 馬場直道, 泉実

    日本農芸化学会大会講演要旨集  2009 

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  • 糸状菌由来の新規β-N-acetylglucosaminidase阻害剤

    仁戸田照彦, 豊岡実穂, 臼木博一, 奥田徹, 神崎浩

    日本農薬学会大会講演要旨集  2007 

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  • 細胞分裂阻害物質phenylahistin生産糸状菌Aspergillus ustus由来のcyclo(Leu-Phe)酸化酵素により変換される新規二次代謝産物

    神崎浩, 杉原孝治, 平田里枝, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集  2006 

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  • 放線菌由来cyclo(Leu-Phe)脱水素酵素の精製と諸性質の検討

    神崎浩, 森本篤史, 池田万里, 仁戸田照彦

    日本農芸化学会中四国支部講演会講演要旨集  2002 

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  • 糸状菌の生産する細胞分裂阻害物質(-)-phenylahistinの培養生産

    神崎浩, 福田隆雄, 仁戸田照彦

    日本農芸化学会大会講演要旨集  2002 

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  • DL-Homocysteine thiolactoneからL-homocysteineへの微生物変換 (第三報)

    神崎浩, 用貝さと子, 藤田恒, 仁戸田照彦

    日本農芸化学会大会講演要旨集  2002 

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  • 微生物で原石を磨く ー天然に存在しない生理活性天然有機化合物の創生ー Invited

    神崎 浩

    日本農芸化学会中四国支部 学会創立 100 周年記念 第 37 回若手シンポジウム (第 13 回農芸化学の未来開拓セミナー)  2023.5.12 

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    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  • 麹菌固体培養による杜仲葉二次代謝産物の微生物変換 (第2報)

    村上響, 奥川日菜乃, 山下秀行, 三木翔平, 仁戸田照彦, 神崎浩

    日本農芸化学会中四国支部第64 回講演会(例会)  2023.1.21 

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  • 糸状菌Pochonia suchlasporia TAMA87株の固体培養による生薬センザンリュウ二次代謝産物の微生物変換

    竹本 実加, 神崎 浩, 仁戸田 照彦

    日本農芸化学会中四国支部第64 回講演会(例会)  2023.1.21 

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  • 麴菌による地域植物資源の高機能化

    神崎 浩

    微生物インダストリー講座開設記念シンポジウム 麴を活かしたものづくり 固体培養が生み出す発酵のイノベーション  2022.11.1 

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    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

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  • 日本が世界に誇る発酵技術による「もの作り」-微生物変換による,医薬品・化粧品・食品機能性素材の開発- Invited

    神崎 浩

    日本科学者会議岡山支部・農学部班合同例会  2021.1.25 

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  • 強力な抗腫瘍活性物質へ変換可能なphenylahistinアナログ,aurantiamineの微生物生産と酵素変換

    日本農芸化学会2007年度大会  2007 

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  • 大腸菌で発現させた放線菌由来Cyclo(Leu-Phe) oxidaseの精製と性質検討

    日本農芸化学会中四国支部第17回講演会  2007 

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  • 微生物変換によるオリーブ成分の高機能化

    2007 

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  • Verticillium sp. F40株が生産する新規 β-N-acetylglucosaminidase阻害剤

    日本農芸化学会2007年度大会  2007 

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  • 微生物変換によるオリーブ二次代謝産物の高機能化

    日本農芸化学会中四国支部第18回講演会  2007 

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  • Pochoina属糸状菌が生産する新規β-N-glucosaminidase阻害剤の阻害活性

    日本農芸化学会2007年度中四国・西日本支部合同大会(中四国支部第19回講演会)  2007 

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  • 昆虫β-N-glucosaminidase特異的阻害剤HU-1の構成異常糖の合成と阻害活性評価

    日本農芸化学会2007年度中四国・西日本支部合同大会(中四国支部第19回講演会)  2007 

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  • Screening for cyclic dipeptide producing microorganisms by cyclo(Leu-Phe) oxidase

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Production of a cell cycle inhibitor phenylahistin using membrane-surface liquid culture

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Purification and charactarization of cyclo(Leu-Phe) oxidase from Streptomyces albulus KO23: An enzyme useful for bioactive dehydro cyclic dipeptide production

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Effect of an acidic polysaccharide PS-1 on Frankia-actinorhizal plant nitrogen-fixing root nodule symbiosis

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Production of 2-O-(b-D-glucopylanosyl)ascorbic acid by cultured cells of Lycium chinese

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Enzymatic synthesis of bioactive dehydrogenated cyclic dipeptides by cyclo(Leu-Phe) oxidase from Streptomyces albulus KO23

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Purification and characterization of L-homocysteine thiolactone hydrolase involved in dynamic resolution process of L-homocysteine.

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • Screening for cyclic dipeptide-producing microorganisms by cyclo(Leu-Phe) oxidase

    日本農芸化学会  2005 

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  • 微生物酵素を利用するものづくり

    日本化学会中国四国支部岡山地区化学講演会  2005 

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  • 放線菌由来環状ジペプチド酸化酵素を利用するCyclo(His-His)脱水素体の合成

    日本農芸化学会  2005 

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  • フッ素アミノ酸添加培養による新規フッ素化アルボノルシンの微生物合成

    生物工学会大会  2005 

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  • Cyclo(Leu-Phe)酸化酵素が触媒するcyclo(Asp(OMe)-Asp(OMe))の脱水素反応

    日本農芸化学会関西・中四国・西日本支部合同支部大会  2005 

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  • Search for enzyme inhibitors specific for insect b-N-acetylglucosaminidase

    2005環太平洋国際化学会議 (PACIFICHEM 2005)  2005 

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  • 微生物酵素を利用するものづくり

    第3回 ニューバイオ技術研究交流会  2005 

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  • 微生物由来酸性多糖のアクチノリザル植物根粒窒素固定促進効果

    日本農芸化学会中四国支部例会  2005 

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  • 昆虫のβ-N-acetylglucosaminidaseに特異的な阻害剤の探索(第3報)

    日本農芸化学会大会  2005 

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  • 抗がん活性化合物の酵素合成

    化学工学会秋季大会 シンポジウム 化学品製造をめざすバイオプロセス  2005 

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  • Novel Bioactive Dehydro Cyclic Dipeptides Prepared by Enzymatic Conversion

    国際微生物学会大会 (IUMS2005)  2005 

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  • 膜面液体培養による細胞周期阻害剤Phenylahistinの生産

    平成16年度日本農芸化学会中四国支部大会  2004 

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  • 硝酸イオン資化性菌のスクリーニングと同定

    日本農芸化学会2004年度大会  2004 

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  • Albonoursin生合成系の素反応 cyclo (Leu-ΔPhe) からalbonoursinへの変換を触媒する酵素に関する検討

    日本農芸化学会2004年度大会  2004 

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  • Cyclo (Gly-Leu)資化性菌が生産するaminopeptidase

    日本農芸化学会2004年度大会  2004 

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  • 糸状菌TNPT116-Cz株が生産する昆虫キチナーゼ阻害多糖FPS-1の阻害特性

    日本農芸化学会2004年度大会  2004 

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  • 昆虫のβ-N-acetylglucosaminidaseに特異的な阻害剤の探索

    日本農芸化学会2004年度大会  2004 

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  • L-Homocysteine thiolactone加水分解酵素の精製と諸性質の検討 (第二報)

    平成16年度日本農芸化学会中四国支部大会  2004 

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  • 微生物由来酸性多糖のアクチノリザル植物根粒形成促進効果

    日本農芸化学会中四国支部第9回講演会  2004 

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  • 生物資源からの高機能物質(生理活性物質)の探索と低機能物質(安価な基質)からの変換

    アグリビジネス創出産学官連携シンポジウム  2004 

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  • 細胞周期阻害剤dehydrophenylahistinのチオカルボニル化

    平成16年度日本生物工学会大会  2004 

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  • 糸状菌TNPT116-Cz株が生産する昆虫キチナーゼ阻害多糖FPS-1の阻害特性

    第8回生体触媒化学シンポジウム  2004 

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  • Agrobacterium radiobacter由来L-homocysteine thiolactone加水分解酵素の精製と性状解析

    第8回生体触媒化学シンポジウム  2004 

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  • 昆虫のb-N-acetylglucosaminidaseに特異的な阻害剤の探索(第2報)

    平成16年度日本農芸化学会中四国支部大会  2004 

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  • 糸状菌TNPT116-Cz株が生産する昆虫キチナーゼ阻害多糖FPS-1の阻害特性(第2報)

    平成16年度日本農芸化学会中四国支部大会  2004 

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  • 昆虫キチナーゼ阻害物質生産糸状菌由来のキチナーゼ

    日本農芸化学会中四国支部,西日本支部,日本栄養・食糧学会西日本支部, 日本食品科学工学会西日本支部 合同大会  2003 

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  • 糸状菌が生産する細胞分裂阻害物質 phenylahistin の生合成

    2003年度日本農芸化学会大会  2003 

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  • 細胞周期阻害剤dehydrophenylahistinの酵素合成 −糸状菌培養物由来の立体異性混合物を基質とする効率的合成−

    日本生物工学会2003年度大会  2003 

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  • L-Homocysteine thiolactone加水分解酵素の精製と諸性質の検討

    2003年度日本農芸化学会大会  2003 

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  • Cyclo(Leu-Phe)からalbonoursinへの酵素変換反応の解析

    日本農芸化学会中四国支部,西日本支部,日本栄養・食糧学会西日本支部, 日本食品科学工学会西日本支部 合同大会  2003 

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  • 放線菌由来cyclo (Leu-Phe)脱水素酵素の性状解析

    日本農芸化学会中四国支部,西日本支部,日本栄養・食糧学会西日本支部, 日本食品科学工学会西日本支部 合同大会  2003 

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  • 糸状菌が昆虫キチナーゼ阻害物質とともに生産するキチナーゼ

    第7回生体触媒化学シンポジウム  2003 

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  • 放線菌由来cyclo (Leu-Phe)脱水素酵素の性状解析

    第7回生体触媒化学シンポジウム  2003 

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  • 放線菌由来環状ジペプチド脱水素酵素系を利用するcyclo(Phe-Pro)脱水素体の合成

    日本農芸化学会中四国支部大会  2002 

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  • 糸状菌HUF45株が生産する昆虫キチナーゼ阻害物質

    2002年度日本農芸化学会大会  2002 

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  • 糸状菌TNPT116-Cz株が生産する昆虫キチナ-ゼ阻害活性を有する多糖

    2002年度日本農芸化学会大会  2002 

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  • 強力な細胞分裂阻害活性を有するdehydrophenylahistinの化学-酵素合成

    2002年度日本農芸化学会大会  2002 

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  • DL-Homocysteine thiolactoneからL-homocysteineへの微生物変換(第三報)

    2002年度日本農芸化学会大会  2002 

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  • 糸状菌の生産する細胞分裂阻害物質(-)- phenylahistin の培養生産

    2002年度日本農芸化学会大会  2002 

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  • 放線菌由来脱水素酵素系を用いた脱水素型環状ジペプチド類の合成

    第6回生体触媒化学シンポジウム  2002 

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  • 放線菌由来cyclo (Leu-Phe)脱水素酵素の精製と諸性質の検討

    日本農芸化学会中四国支部大会  2002 

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  • Agrobacterium属細菌の凝集促進活性を有する微生物細胞外多糖の性状解析

    日本農芸化学会中四国支部大会  2002 

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  • 環状ジペプチド分解細菌が生産するジペプチダーゼの精製と諸性質の検討

    2002年度生物工学会大会  2002 

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  • 放線菌由来環状ジペプチド脱水素酵素系を利用するCyclo (Met-Met)の脱水素体の合成

    日本農芸化学会2001年度関西・西日本・中四国支部合同大会  2001 

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  • DL-ホモシステインチオラクトンからL-ホモシステインへの立体選択的微生物変換

    第4回生体触媒化学シンポジウム  2001 

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  • 生物変換による新規抗腫瘍活性化合物の生産

    バイオプロセスによる有機合成 岡山ミニシンポジウム(第4回)  2001 

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  • 糸状菌の生産する昆虫キチナーゼ阻害物質

    日本農芸化学会2001年度大会  2001 

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  • Novel Diketopiperazine Metabolism in a Microorganism

    18th International Congress on Heterocyclic Chemistry (第18回国際複素環化合物会議)  2001 

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  • Production of Novel Bioactive Dehydro Cyclic Dipeptides By Microbial Enzyme System

    18th International Congress on Heterocyclic Chemistry (第18回国際複素環化合物会議)  2001 

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  • 放線菌由来の新規環状ジペプチド脱水素酵素系を利用する有用物質生産(第二報)

    日本農芸化学会2001年度大会  2001 

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  • Cyclo(Leu-Phe) からalbonoursin への脱水素反応を触媒する新規酵素系(第五報)

    日本農芸化学会2001年度大会  2001 

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  • 脱水素型環状ジペプチド生合成酵素系による生理活性物質生産

    第43回天然有機化合物討論会  2001 

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  • Production of Novel Bioactive Dehydro Cyclic Dipeptides By Microbial Enzyme System

    Biotrans 2001(The 5th International Symposium on Biocatalysis and Biotransformation)  2001 

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  • 糸状菌由来昆虫キチナーゼ阻害物質の探索および性状比較

    日本農芸化学会2001年度関西・西日本・中四国支部合同大会  2001 

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  • 酵素による生理活性物質生産と情報化学の接点

    藪田セミナー「IT時代のバイオサイエンス」  2001 

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  • 糸状菌由来昆虫キチナーゼ阻害物質の探索

    第5回生体触媒化学シンポジウム  2001 

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  • 生理活性を示す脱水素環状ジペプチド生産に有用な放線菌由来の新規脱水素酵素系

    第5回生体触媒化学シンポジウム  2001 

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  • Agrobacterium tumefaciensの植物形質転換に関与する多糖

    日本農芸化学会2001年度関西・西日本・中四国支部合同大会  2001 

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  • Cyclo(Gly-Leu)資化性菌由来Leucylglycine分解酵素

    第5回生体触媒化学シンポジウム  2001 

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  • Synthesis of bioactive dehydrodiketopiperazines catalyzed by a novel actinomycetous enzyme system

    Pacifichem 2000  2000 

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  • cyclo(Leu-Phe) から albonoursin への脱水素反応を触媒する新規酵素系(第四報)

    日本農芸化学会2000年度大会  2000 

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  • 放線菌の新規酵素系を用いた細胞分裂阻害活性を示す脱水素型環状ジペプチド類の合成

    日本農芸化学会2000年度大会  2000 

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  • Agrobacterium属細菌の植物への付着を阻害する oxazolomycin diacetate の作用点

    日本農芸化学会2000年度大会  2000 

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  • 放線菌新規酵素系を用いる生理活性脱水素型ジケトピペラジン類の合成

    第3回生体触媒化学シンポジウム  2000 

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  • 糸状菌の生産する昆虫キチナーゼ阻害物質

    日本農芸化学会2000年度大会  2000 

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  • cyclo(Gly-Leu)資化性菌が生成するdipeptidase

    日本農芸化学会2000年度大会  2000 

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  • 微生物細胞外酸性多糖の植物形質転換に対する生理活性

    日本農芸化学会2000年度関西支部大会  2000 

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  • 微生物由来のジケトピペラジン環化合物代謝酵素による生理活性物質生産

    日本生物工学会平成12年度大会  2000 

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  • Production of Novel Bioactive Dehydro Cyclic Dipeptides by Streptomyces Enzyme System

    『耐熱性微生物の学際的研究ならびに資源開発とその応用』に関する拠点大学方式によるタイ国との共同研究の第2回合同セミナー  2000 

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  • 放線菌由来の新規環状ジペプチド脱水素酵素系を利用する有用物質生産

    日本農芸化学会2000年度関西支部大会  2000 

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  • 環状ジペプチド, cyclo (Gly-Leu) の微生物分解代謝に関与する新規酵素系

    日本生物工学会平成12年度大会  2000 

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  • Novel microbial metabolism of a morphine antagonist cyclo (Gly-Leu)

    Pacifichem 2000  2000 

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  • A Search for Insect Chitinase Inhibitors of Fungal Origin

    Pacifichem 2000  2000 

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  • Oxazolomycin diacetate, inhibitor against agrobacterial attachment to plants

    Pacifichem 2000  2000 

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  • Cyclo(Phe-Leu) からalbonoursin への脱水素反応を触媒する新規酵素系(第二報)

    日本農芸化学会1999年度大会  1999 

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  • デヒドロアミノ酸を構成成分とするジケトピペラジン類の新規生合成系を利用する生理活性物質の酵素合成

    日本放線菌学会 1999年度大会  1999 

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  • Oxazolomycin ester の示すAgrobacterium 属細菌の植物への付着阻害活性

    日本農芸化学会1999年度大会  1999 

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  • DL-Homocysteine thiolactone から L-homocysteine への微生物変換(第二報)

    日本農芸化学会1999年度大会  1999 

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  • 微生物による cyclo(Gly-Leu) の分解

    日本生物工学会1999年度大会  1999 

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  • 寄付講座『微生物インダストリー講座』開設記念シンポジウムを開催

    神崎 浩, 深野 夏暉, 藤原 加奈, 狩山 昌弘

    岡山大学 教育研究等の状況の広報に係る記者発表 

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    Language:Japanese   Presentation type:Media coverage  

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Industrial property rights

  • 脂肪蓄積抑制剤及びその製造方法並びに血糖値上昇抑制剤及び腸内細菌叢改善剤

    神崎 浩, 仁戸田 照彦, 畑生 俊光

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    Applicant:国立大学法人 岡山大学

    Application no:特願2020-004396  Date applied:2020.1.15

    Announcement no:特開2021-109865  Date announced:2021.8.2

    J-GLOBAL

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  • 抗酸化性化合物及びそれを含有する組成物の製造方法、並びに、それに用いられる新規微生物

    神崎 浩, 仁戸田 照彦, 徐 恵美, 菊地 敬一

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    Applicant:国立大学法人 岡山大学

    Application no:特願2015-139849  Date applied:2015.7.13

    Announcement no:特開2017-018051  Date announced:2017.1.26

    Patent/Registration no:特許第6609132号  Date registered:2019.11.1 

    J-GLOBAL

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  • オリーブ葉抽出物を酵母処理することにより得られる抗酸化物質

    神崎 浩, 仁戸田 照彦, 服部 恭一郎, 吉田 靖弘

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    Applicant:国立大学法人 岡山大学

    Application no:特願2007-039296  Date applied:2007.2.20

    Announcement no:特開2008-201715  Date announced:2008.9.4

    Patent/Registration no:特許第4652355号  Date registered:2010.12.24 

    J-GLOBAL

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  • N-アセチルグルコサミニダーゼ阻害活性を有する化合物

    神崎 浩, 仁戸田 照彦, 奥田 徹, 内田 有紀

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    Applicant:国立大学法人 岡山大学

    Application no:特願2007-035676  Date applied:2007.2.16

    Announcement no:特開2008-195696  Date announced:2008.8.28

    Patent/Registration no:特許第4636623号  Date registered:2010.12.3 

    J-GLOBAL

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  • β-N-アセチルグルコサミニダーゼ阻害活性を有する化合物

    神崎 浩, 仁戸田 照彦, 臼木 博一, 小村 啓, 山路 奈保子

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    Applicant:サントリーホールディングス株式会社

    Application no:特願2005-364111  Date applied:2005.12.16

    Announcement no:特開2007-161690  Date announced:2007.6.28

    Patent/Registration no:特許第5049489号  Date registered:2012.7.27 

    J-GLOBAL

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  • 細胞分裂阻害剤及びその製造方法

    神崎 浩, 加納 周雄, 柳澤 恵広, 仁戸田 照彦, 赤澤 和美

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    Applicant:ネリアス・ファーマシュティカルズ・インコーポレーテッド

    Application no:特願2001-553764  Date applied:2000.9.29

    Patent/Registration no:特許第4810043号  Date registered:2011.8.26 

    J-GLOBAL

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Awards

  • 日本生物工学会斎藤賞

    2000  

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Research Projects

  • タイ産有色米の麹菌固体培養微生物変換による高機能化

    2023.03 - 2024.03

    飯島藤十郎記念食品科学振興財団  2022年度外国人留学生研究助成 

    Jitpakdee Jirayu, 神崎 浩

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  • Microbial conversion of plant resources by three different controlled Koji-mold solid cultivation methods

    Grant number:22H02242  2022.04 - 2026.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    神崎 浩, 仁戸田 照彦

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    Grant amount:\17420000 ( Direct expense: \13400000 、 Indirect expense:\4020000 )

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  • 麹菌固体培養技術を駆使する高機能化日 本ワインパミス麹の創生

    2021.07 - 2022.03

    岡山工学振興会  令和3年度学術研究助成 特別研究 内山勇三 科学技術賞 

    神崎浩, 仁戸田照彦, 伊藤一成, 谷野有佳, 竹内赴登

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    Authorship:Principal investigator 

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  • 食品・化粧品素材開発を目指す麹菌固体培養によるオリーブ資源の高機能化

    2021.04 - 2022.03

    大下財団  2021年度大下財団「生物機能の産業利用・工業化」研究助成 

    神崎浩, 仁戸田照彦

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  • 2020年度 小型通気式固体培養装置を用いる麹菌による地域植物資源の 高機能化 補助事業

    2020.04 - 2021.03

    JKA  JKA 機械振興補助事業 開発研究 

    神崎浩, 仁戸田照彦

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  • 日本ワインぶどう果皮成分の麹菌固体培養による食品・化粧品機能性素材への高機能化

    2020.04 - 2021.03

    中国地域創造研究センター  2020年度 新産業創出研究会 

    神崎 浩, 仁戸田 照彦, 三宅 剛史, 伊藤 一成, 谷野 有佳, 山下 秀行, 中川 拓郎, 平野 幸司, 栢菅 一久

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  • サブサイト認識阻害剤による新奇β-N-アセチルグルコサミニダーゼの探索と性質精査

    Grant number:19K05792  2019.04 - 2022.03

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    神崎 浩

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    糖質加水分解酵素(GH)には同じファミリーに属しながら, Subsiteによる基質認識機構が異 なる酵素が知られている。GHの基質認識機構の多様性を詳細に明らかにするには,ユニークな基質認識機構を有する新奇なGHを見出すことが重要である。 我々は,β-N-Acetylglucosaminidaseの阻害剤TMG-chitotriomycinを発見し、TMG(Trimethylglucosaminium)残基が阻害活性に重要で、PNP(p-Nitrophenyl)-TMGも阻害を示すことを明らかにした。 今回、TMG阻害剤に対して既存酵素と異なる阻害特異性を示す酵素を生産する微生物の獲得を目指した。
    自然界から単離したPNP-GlcNAc資化性菌27株を,ブイヨン培地およびpNP-GlcNAc培地で培養し,休止菌体のPNP-GlcNAc分解活性を測定したところ、ブイヨン培地を用いて培養したとき,全ての菌株においてpNP-GlcNAc分解活性が確認されたことからGlcNAcaseを構成的に発現していると推測された。
    また、両培地でβ-GlcNAcaseを高生産するMK26株の, PNP-GlcNAc含有培地で生産される酵素(β-GlcNAcase-M K26-pGN)は,ブイヨン培地で生産される酵素(β-GlcNAcase-MK26-Bouillon)と異なり,タンパク質の疎水性が高く,界面活性剤存在下で抽出液 中に活性が回収された。1つの菌株が2種の 特性の異なるβ-GlcNAcaseを生産することは大変興味深い。
    β-GlcNAcase-MK26-Bouillonについて、イオン交換及びゲル濾過クロマトグラフィー で部分精製し、その部分精製酵素についてTMG-chitotriomycin、PNP-TMGに対する阻害特異性を検討したところ、既存酵素と異なっていることが判明した。

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  • オリーブ葉成分の微生物変換による高機能化と化粧品素材としての応用開発研究

    2019

    日本農芸化学会  農芸化学中小企業産学・産官連携研究助成金 

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  • Subsite studies on active sites of beta-diglycoside-hydrolyzing enzymes based on detail substrate specificity analysis

    Grant number:15K07393  2015.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KANZAKI HIROSHI, SUGANUMA Shoko, KOIDE Mana

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    Grant amount:\5070000 ( Direct expense: \3900000 、 Indirect expense:\1170000 )

    In the case of β-Glucosidase studies, we purified three different pNPG-hydrolyzing enzymes (50kDa, 102kDa, 117kDa) from the enzyme preparation of Penicillium multicolor and found that the 102kDa enzyme exhibited the unique substrate specificity. In the case of GlcNAcase studies, microorganisms assimilating pNP-TMG, an analog of TMG-chitotriomycin, were isolated, and some of them exhibited pNP-TMG- and pNP-GlcNAc-hydrolyzing activities. As pNP-TMG was found to inihibit the known GlcNAcase, elucidation of the substrate recognition mechanism of the enzymes of pNP-TMG-asimilating microorganisms is supposed to be interesting.

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  • 酵素変換による高感度検出法を用いる微生物由来新規生理活性環状ジペプチド類の探索

    Grant number:07F07431  2007 - 2009

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    神崎 浩, BUETUSIWA M T, BUETUSIWA M. T.

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    Grant amount:\2300000 ( Direct expense: \2300000 )

    放線菌Streptomycs albulus KO23株が生産する環状ジペプチド酸化酵素(CFLoxidase)が環状ジペプチドに対して,非常に幅広い特異性を示すこと,その反応生成物,脱水素型環状ジペプチドが特徴的なUVスペクトルを示すことから,酵素反応とDAD-HPLC法を組み合わせることで、環状ジペプチドを効率よく検出する方法を確立した。平成19年度の研究において,その検出法をより簡便で高感度にする手法へと改善し,平成20年度はこの方法を用いて,微生物および植物抽出物を探索源として環状ジペプチドの検索を行い,幾つかの抽出物中に酵素反応を受ける化合物の存在を認めた。
    今年度はまず,環状ジペプチドと予測される化合物の存在を認めた数種の糸状菌抽出物のうち,鰹節生産に用いられる糸状菌Aspergillus repensの培養抽出物から,目的化合物を単離し,脱水素型環状ジペプチドneoechinilin Aと同定した。さらにこの化合物を基質としたCFLoxidaseによる変換反応で得られるneoechinilin Bを単離した。これらの化合物の細胞分裂阻害活性を測定し,環状ジペプチド類の構造活性相関に関して新たな知見を得た。すなわち,これらの化合物に共通するTrp残基に置換しているイソプレニル基が,細胞分裂阻害活性の発現に重要であることが判明した。また,CFLoxidaseによる酵素反応を受ける化合物の存在を認めたマツ培養細胞抽出物から,目的化合物の単離精製に成功した。

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  • Confirmation of multiple symbiotic systems of plant-Frankia-mycorrhiza with a higher stress tolerance and its application to the rehabilitation of degraded lands

    Grant number:16380051  2004 - 2007

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    SASAKAWA Hideo, KANZAKI Hiroshi, YAMAMOTO Mikihiro, SHIMA Kazuto, EZAWA Tatsuhiro

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    Grant amount:\13010000 ( Direct expense: \12500000 、 Indirect expense:\510000 )

    This research was clone to establish a multiple symbiotic system of actinorhizal plant-Frankia- mychorrhiza with a higher environmental stress tolerance and to confirm it as an effective system for the recovery of the degraded lands. The results obtained were as follows.
    1. One strain from the root nodules of Elaeagnus macrophylla and 4 strains from the root nodules of Myrica rubra were isolated and they were identified as Frankia by their morphological characteristics and inoculation test.
    2. Frankia Apel strain was isolated from the root nodules of Alnus pendula growing in acid soil. The growth pH range of the strain was between 6.0 and 8.0, and the value was not different from those of Frankia which were isolated from the root nodules of actinorhizal plants growing in neutral pH.
    3. Host range of each Frankia strain obtained by inoculation test and the result of phylogenic analysis with 16S rDNA sequence of each strain were well consistent.
    4. Frankia strains reserved in the lab could be well discriminated by RFLP analysis of nifD-KIGS region and this method was effective for the determination of Frankia strain in the nodules of actinorhizal plants growing under natural conditions.
    5. Pisolithus tinctorius or Cenococcum geophilum which had been isolated and reserved were inoculated with Frankia to 4 actinorhizal plants, but no mycorrhizal infection was observed.
    6. One mycorrhiza-like fungus was isolated from 4 species of pine mycorrhizal mushrooms, respectively, and also one mycorrhiza-like fungus was isolated from mycorrhizal mushroom of Alnus sieboldiana. PCR-RFLP analyses strongly suggested that these fungi could be mycorrhizal fungi which infect on both pine and Alnus species.
    These results gave us a prospect for the establishment of tripartite symbiotic system on actinorhizal plants which is effective for the recovery of the degraded lands

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  • コンビナトリアルバイオコンバージョンによる新規生理活性物質ライブラリー創製の試み

    Grant number:14656046  2002 - 2003

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    神崎 浩, 仁戸田 照彦

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    Grant amount:\3400000 ( Direct expense: \3400000 )

    本来生物が生産しない化合物を生物変換で合成する研究は,生理活性物質生産において幾つか知られている。しかしこのような反応は,目的物が決まった上で選抜されたものであり,ランダムにバイオコンバージョンを行い,その結果得られる化合物群をラィブラリーとする研究はほとんど行われていない。現在の医薬などの生理活性物質探索では,ハイスループットスクリーニング(HTS)が全盛であり,そのための化合物ライブラリーの構築が不可欠とされている。そのためにコンビナトリアル合成が行われているが,これもあくまで構造予測のうえでのライブラリー構築である。そこで,本研究では,2種類の生物の持つ二次代謝産物生産能を組み合わせて,1種の生物では生産不可能な化合物を合成する『コンビナトリアルバイオコンバージョン』という手法の基礎的な検討を行った。我々は既に,糸状菌Aspergillus ustusが生産する細胞分裂阻害物質phenylahistinを放線菌Streptomyces albulusが有している環状ジペプチド脱水素酵素系で処理すると,出発物質のさらに1000倍以上も強力な活性を有する新規化合物dehydrophenylahistinが生成することを明らかにしており,昨年度は,この反応をより効率よく行う技術開発を試み,2種の微生物を寒天培地上で培養し,途中で微生物を移し替える培養法により,目的物質の生産が可能となった。本年度は寒天培養より様々な利点を有する液体培養による方法の確立を試みた。液体振とう培養や,液体静置培養ではPhenylahistinの生産はほとんど見られなかったこと,菌の移し替えを考えると膜状で菌体を生育させるのが望ましいことから,岡山大学工学部中西教授が考案された膜面液体培養による生産を試みたところ,寒天培養より生産性の高い条件が確立できた。以上のことから,寒天培養あるいは膜面液体培養で二種の微生物を培養し,培養途中にそれらを移し替える二段階醗酵により,新規の化合物が生産できると期待される。

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  • Development of reagents for effective genetic modification of plants

    Grant number:09556024  1997 - 1999

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    KANZAKI Hiroshi, OBATA Tokio, NITODA Teruhiro

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    Grant amount:\11400000 ( Direct expense: \11400000 )

    Tow novel geometrical isomers of oxazolomycin, oxazolomycins B (2) and C(3), were isolated from the fermentation broth of an oxazolomycin- producing strain, Streptomyces albus JA3453. The configurations of the triene moieties of 2 and 3 are (4'E, 6'E, 8'E) and (4'Z, 6'E, 8'E), respectively, while that of oxazolomycin (1) is (4'Z, 6'Z, 8'E). 2 and 3 exhibited potent inhibitory activity against crown gall formation with the same MIC (0.8 μg/disk) as oxazolomycin. 2 and 3 showed no antibacterial activity against Agrobacterium tumefaciens, in contrast to oxazolomycin which has specific anti-A. tumefaciens activity. The newly established GUS expression bioassay of the plant callus extracts revealed that the extracts of Phytolacca americana callus had the most potent inhibitory activity against agrobacterial plant transformation. A triterpene glycoside phytolaccoside B was isolated from the extract as a genuine plant transformation inhibitor having neither antiagrobacterial nor phytotoxic activity. We found that the inhibitory activity of oxazolomycin oxazolomycin diacetate against plant transformation was due to its inhibitory effect against the attachment of A. tumefaciens to host plants. Furthermore, oxazolomycin diacetate was found to exhibit an inhibitory activity against the rhicadhesin-mediated agrobacterial attachment to plants, not the biosynthesis of cellulose and β-1, 2-glucan which have been reported to be involved in the agrobacterial attachment. And we found that a novel bacterial acidic polysaccharide from a soil bacterium exhibited plant transformation enhanced plant transformation due to its stimulatory effect for the attachment of A. tumefaciens to host plants.

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  • Analysis of biosynthetic enzyme system for dehydrodiketopiperazines with antitumor activity and its application

    Grant number:09660095  1997 - 1999

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KANZAKI Hiroshi

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    Grant amount:\3300000 ( Direct expense: \3300000 )

    Albonoursin production was greatly enhanced when cyclo (L-Leu-L-Phe) (CFL), a tetrahydroderivative of albonoursin, was added to the culture of an albonoursin-producing actinomycete. Furthermore, the resting cells and the cell-free extract of the strain catalyzed the bioconversion of CFL to albonoursin. We isolated and identified the biosynthetic intermediates of this conversion as (Z)-3-benzylidene-6-isobutyl-2, 5-piperazinedione and (Z)-3-benzyl-6-isobutylidene-2, 5-piperazinedione. These results revealed that albonoursin was biosynthesized by dehydrogenation of CFL in the actinomycete. We established the simple assay method for this enzyme activity using phenazine methosulfate and dichlorophenol indophenol as hydrogen acceptors. The cell-free extract was found to catalyze the conversion of several cyclic dipeptides other than CFL to the corresponding dehydro derivatives. Thus, this system might be a powerful tool for producing a variety of dehydro derivatives. Among dehydro cyclic dipeptides prepared, tetradehydro derivatives exhibited an inhibitory activity toward the first cleavage of sea urchin embryo, while didehydro derivatives did not. These facts indicated that dehydrogenation at α, β-positions of both amino acid residues in this type of cyclic dipeptides were required for the inhibitory activity.

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  • 微生物が生産するジケトピペラジン環分解酵素の性質と物質生産への利用

    Grant number:06760088  1994

    日本学術振興会  科学研究費助成事業 奨励研究(A)  奨励研究(A)

    神崎 浩

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    Grant amount:\1000000 ( Direct expense: \1000000 )

    ジケトピペラジン環構造を有する化合物は植物や微生物の二次代謝産物中に多く見いだされており,抗癌活性・抗菌活性などの興味深い生理活性を有している。本研究者は,ジケトピペラジン環が基本的に2分子のアミノ酸が脱水縮合した構造であること,プロテアーゼの逆反応によりペプチドの合成が可能となっていることから,ジケトピペラジン分解酵素が取得できれば,その酵素の多機能性を利用してジケトピペラジンが合成可能であると推測し,ジケトピペラジン分解酵素の探索を行ってきた。ジケトピペラジン分解酵素としては無水グリシン分解酵素のみが報告されているが,その酵素の基質特異性は非常に高く,無水グリシン以外を基質としないため,物質生産には必ずしも適当とは言えない。この無水グリシン分解酵素が菌体外酵素であるのに対しこれまでに本研究者は菌体内に無水グリシン分解活性を有する菌株を取得している。今回,無水グリシルチロシン,無水フェニルアラニルロイシンの2種を資化する微生物を土壌から単離し,それらの分解酵素生産菌のスクリーニングを行った。その過程で菌体内に無水グリシルチロシン分解活性を有する菌株を見いだし,菌体反応によりジケトピペラジン類に対する基質特異性を検討したところ,無水チロシルグリシン以外に多くのジケトピペラジン類が分解された。片方のアミノ酸をグリシンで固定し,もう一方を中性アミノ酸で変化させたところ,かさ高さが大きいアミノ酸がよりよい基質となった。また,D-体のアミノ酸由来のジケトピペラジンはほとんど基質とならないことが分かった。一方,無水フェニルアラニルロイシンは水に対する溶解性が極端に低いため,その分解酵素誘導のために2段階培養を採用した。すなわち,一般の細菌用培地で充分に菌を生育させた後,無水フェニルアラニルロイシンを含む培地に無菌的に移すことにより酵素誘導を行わせた。この条件で菌体外に分解活性を示す菌株を取得した。この方法により,他のジケトピペラジン分解酵素も容易に大量に誘導可能と考えられる。今後,菌体外の酵素を精製し,その諸性質を明らかにすることにより,ジケトピペラジン類の酵素合成の可能性を追及する予定である。

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  • 生理活性ジケトピペラジン分解酵素の性質及びその応用

    Grant number:04760081  1992

    日本学術振興会  科学研究費助成事業 奨励研究(A)  奨励研究(A)

    神崎 浩

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    Grant amount:\900000 ( Direct expense: \900000 )

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  • Organic chemial approach to the elucidation of the mechanism of plant transformation induced by Agrobacterium tumefaciens

    Grant number:03453145  1991 - 1993

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for General Scientific Research (B)  Grant-in-Aid for General Scientific Research (B)

    KAWAZU Kazuyoshi, KANZAKI Hiroshi, KOBAYSHI Akio

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    Grant amount:\6900000 ( Direct expense: \6900000 )

    Plant transformation by Agrobacterium tumefaciens has been dramatically developed to create plants acquiring new characters, e.g., herbicide- or virus-resistance. Many scientists have been exploring the mechanism for the plant transformation, although the entire mechanism is not yet fully understood. As the plant transformation consists of several complex steps, a specific chemical probe affecting each step is required to elucidate its mechanism. The discovery of transformation regulators and the classification of these probes based on their mode of action are both helpful. We devised a rapid and quantitative bioassay method for regulators of plant tranformation by A.tumefaciens with an intron-GUS binary vector as an alternative method to the time-consuming potato tuber disk assay. Among several host plants tested, tobacco BY-2 suspension cells were the most preferable for this GUS expression assay. Furthermore, as the phytotoxicity of the test material, if any, is apparent from the growth inhibition of BY-2 suspension cells, this method can exclude phytotoxic compounds.
    In the course of our screening by the potato tuber disk assay for inhibitors of crown gall formation, oxazolomycin was isolated. Oxazolomycin inhibited crown gall formation owing to strong anitbacterial activity to the pathogen, A.tumefaciens. Interestingly, oxazolomycin esters had no antibacterial activity, although they inhibited crown gall formation. Curromycins, oxazolomycin analogs, were reproted as antibiotics. We isolated curromycin B from the curromycin-producing strain and prepared curromycin B diacetate. Curromycin B showed both crown gall formation inhibitory and antibacterial activities against A.tumefaciens. Curromycin B diacetate did not show any antibacterial activity, although it inhibited crown gall fromation. These results indicated that curromycin B diacetate was a promising chemical probe for studying the mechanism of plant transformation by A.tumefaciens.

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  • Development of trunk injection agents for a pine-wilt desease

    Grant number:03556016  1991 - 1992

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Developmental Scientific Research (B)  Grant-in-Aid for Developmental Scientific Research (B)

    KAWAZU Kazuyoshi, KANZAKI Hiroshi, KOBAYASHI Akio

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    Grant amount:\8300000 ( Direct expense: \8300000 )

    Dehydrosinefungin is the potent nematicidal compound, but the productivity of this compound by Strepromyces sp. H-114 is very low. Thus, an improvement of the productivity is needed. We developed the quantitative method for dehydrosinefungin to find the best conditions for production of this compound. Paired-ion chromatography using an HPLC aparatus resulted in
    the best separation of dehydrosinefungin and its analogs, sinefungin and amidinosinefungin. Guided by the productivity of dehydrosinefungin, we determined the best culture conditions. Using the best conditions, the productivity reached more than 20 mg/l, that is, ten times as high as that of original conditions. Large scale cultivation using a jar-fermentor gave us more than 200 mg of pure dehydrosinefugin. This enabled us to carry out the assay using three-year pine trees.
    On the other hand, we established the method for a trunk injection of the active compound using morantel tartarate as a test material. Using this method, dehydrosinefungin was injected to three-year pine trees before the treatment of nematodes. We found that 0.5 mg dehydrosinefungin per a three-year pine tree resulted in the resistance of tested trees against wilting. This result indicated that dehydrosinefungin was the promising agent for a pine-wilt desease.

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  • シンク器官形成を誘導・促進する生理活性物質の探索分離

    Grant number:03257205  1991

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    小林 昭雄, 神崎 浩

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    Grant amount:\1500000 ( Direct expense: \1500000 )

    天然に存在する多糖類には中性,塩基性多糖類があり,構成する単糖の種類によってその機能が異なる。シンク細胞あるいはソ-ス細胞の成立には種々の要因が考えられるが,近年細胞膜由来の微量なオリゴ糖の関与が指摘されている。このような状況の中で昨年度以来中性多糖であるデンプン,セルロ-ス,カ-ドラン,ペクチン,ラミナラン等から,また塩基性多糖であるキチン,キトサン等から,酵素あるいは酸分解によってオリゴ糖を切り出し,その断片のもつ二次代謝産物合成経路の活性化に対する影響を検討した。そこで天然多糖をモデルとし,アルファルファ子葉のフラボノイド生合成経路の活性化を4',7ーdihyd roxyflavone ,4',7ーdihydroxyflavanone,medicarpinの蓄積量を指標とし評価した。最も活性の高かったEisenia bicyclis由来のラミナランをTFAにより加水分解したところ,活性が著しく増大した。このことから加水分解により活性単位が切り出されることが示唆された。そこでこれを活性炭カラムに供したところ80%MeOH溶出区に活性が認められ,それをBioーGe1 Pー4カラム,逆相HPLCにより分取を行い,活性画分を得た。これをピリジルアルミノ化し,LCーMSに供したところ質量1070と1232に相当するイオンピ-クがそれぞれ多数確認できた。即ち,この活性画分は結合様式の異なった6糖と7糖の混合物からなることが明らかとなった。また,カビ,放線菌細胞壁を構成する塩基性多糖キチンを濃塩酸で加水分解し,キチン6糖を調制し還元アミノ化反応により還元末端にフェノ-ル性化合物チラミンを導入した。これをアルカリ部分脱アセチル化し,エンドウに対するエリシタ-活性をファイトアレキシンであるpisatinの蓄積量を指価したところ,これまでエンドウ・エリシタ-と考えられていたキトサン6糖より2.4倍の活性を示した。
    LCーMS分析により本オリゴ糖は2脱アセチルキチン6糖であることが明らかとなった。

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  • シンク器官形成を誘導・促進する生理活性物質の探索分離

    Grant number:02261213  1990

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    小林 昭雄, 神崎 浩

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    Grant amount:\2000000 ( Direct expense: \2000000 )

    天然多糖の潜在的エリシタ-活性の開拓とその組織培養系,特に研究が進んでいない脱分化初期組織を用いた有用物質生産に対する応用を目的としてマメ科植物をモデルに脱分化初期組織におけるシンク細胞からソ-ス細胞への相互変換機構を解析した。
    1.アルファルファ子葉組織に含まれる蛋白質を脱分化後経時的にSDSーPAGEにより分析すると0〜7日目にかけて顕著な変動が見られ,それ以降は泳動パタ-ンに変化はみられなかった。子葉組織を脱分化後経時的にG1cβ→3G1cβ1→6G1c結合を持つラミナラン水溶液(500μg/m1,48hr)に浸漬し,誘導されるフラボノイドの細胞内蓄積量・細胞外排出量をHPLC分析した。その結果,メディカルピンが誘導されたほか,生合成的に初期段階に位置する4',7ーdihydroxyflavanoneと4',7ーdihydroxyflavoneに蛋白質変動に対応した顕著な変化がみられた。両化合物は0〜5日目に著し誘導された。しかし7日目以降はほとんど誘導されなかった。フラバノンは細胞内蓄積量に比して細胞外排出量が約3倍を示し,フラボンは誘導初期(0〜3日目)は細胞外に排出されるが,5日目には細胞内に保持された。
    2.G1cNAcβ1→4G1cNAc結合を持つキチンをキチン資化性放線菌菌体で処理することにより得られたキチン分解物(100μg/m1,24hr)およびG1cNβ1→4G1cN結合を持つキトサン水溶液(4mg/m1,24hr)でエンドウ上胚軸組織を脱分化後経時的に処理し,誘導されるフラボノイド蓄積量をHPLCで定量した。6日目には母植物を大幅に上回るピサチンの蓄積が認められた。4〜12日目にかけてピサチンと酷似したUVスペクトルを示す成分の顕著な蓄積が見られた。この成分を糖断片で刺激した脱分化6日目のエンドウ上胚軸から精製・単離し構造解析したところ,報告例のない2ーhydroxypisatinであった。

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  • 土壌微生物由来、新規微小管配向制御物質の性状解析と作用機作

    Grant number:02660142  1990

    日本学術振興会  科学研究費助成事業 一般研究(C)  一般研究(C)

    小林 昭雄, 神崎 浩

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    胚細胞が最初のM期を迎えるまでに,雌核雄核の融合,中心体の複製,DNA合成,染色体の凝集など一連の生化学反応が完結する必要があり,この反応系に微小管からなる細胞骨格が重要な役割を果たしている。
    我々は有糸分裂を最も効果的に阻止するには,分裂装置形成を特異的に阻害するのが有効と考え,現在までにTー1をはじめとする新しい有系分裂停止物質を明らかにしてきた。今回,偏光及び蛍光顕微鏡を用いて,カシパンウニの中期紡錘体を解消させ,倍数核を誘起する新しい活性をもった化合物を微生物代謝産物中に求めた。
    さまざまな環境の土壌より放線菌を分離し,10mlのPSM培地で振盪培養した後アッセイに供した。その結果,KAー48株に目的とする活性を認めた。すなわち,中期紡錘体を本培養液で処理すると,紡錘体は直ちに解消した。このことより,本培養液は微小管脱重合活性を持つことが示唆された。また,受精20分後にウニ胚を処理し,2時間後に核をDAPI染色したところ大きな核が認められ,倍数性の誘導が確認された。このことは本活性物質が微小管に特異的に作用しており,DNA合成及び呼吸に影響を与えない事を示唆している。そこで,放線菌KAー48株の生産する活性物質を明らかにするため,大量培養を行い活性物質の単離精製を試みた。
    放線菌KAー48株をPSM培地(3l)で27℃,5日間振蕩培養し,濾過後培養液を酢酸エチルで分配した後,シリカゲルカラムクロマトグラフィ-及びODSカラムクロマトグラフィ-で精製し3種の活性物質を得た。
    これらはsilica TLC上,Rf0.47,0.30(BenzeneーAcetone=8:2),Rf0.25(benzeneーacetoneーMeOH,7:2:1)のスポットとして確認できた。また,これら化合物はいずれも330nm付近にUVの極大吸収を示した。構造の詳細については現在検討中である。

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  • Chemical Studies on the Infection Mechanism of Crown Gall Disease

    Grant number:01560152  1989 - 1990

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for General Scientific Research (C)  Grant-in-Aid for General Scientific Research (C)

    KAWAZU Kazuyoshi, KANZAKI Hiroshi

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    Grant amount:\2100000 ( Direct expense: \2100000 )

    1. Agrobacterium tumefaciens is a soil bacterium which induces crown gall tumors in a wide variety of dicotyledonous plants. This tumorigenic interaction between the bacterium and the plant is a natural example of the genetic engineering of eukaryotes. Indeed, the oncogenic bacterium harbors a large Ti plasmid, whose T-DNA region is transferred to and integrated into the plant genome during tumorous transformation. However, very little information is available concerning the early process leading to tumor formation. In order to study further the mechanism of tumor induction, it is necessary to develop a chemical probe that will help in exploring the early steps of tumor formation. We searched transformation inhibitors which do not inhibit the growth of A. tumefaciens and are nontoxic against the transformed plant cells.
    2. A few esters derived from oxazolomycin isolated from Streptomyces sp. KBFP-2025 as well as two active compounds of bisanthraquinone type, julichrome Q_<1・3> and julimycin B-II, isolated from Streptomyces sp. TM-71, inhibit crown gall formation, but are nontoxic against this bacterium and the host plant.
    3. These compounds administered to a potato disk 12 - 24 hours after the inoculation of the bacterium hardly inhibited crown gall formation, and no longer necrosed the potato disk. This suggests that these compounds act on any step of transformation process, but have no toxicity toward transformed plant cells whose genome has integrated the T-DNA of A. tumefaciens. It is probable that the transformation process of this system is completed within 12 - 24 hours after the inoculation. For a direct proof of this hypothesis, a quantitative microanalytical method of opines, indices of transformation, has to be established. This method involves HPLC analysis of fluorescent compound formed by reaction of benzoin with guanidino group of the characteristic amino acid. Time course of transformation will be possibly traced by this method.

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  • Microtubulin assembly inhibitors from microorg origin

    Grant number:63560130  1988 - 1989

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for General Scientific Research (C)  Grant-in-Aid for General Scientific Research (C)

    KOBAYASHI Akio, KANZAKI HIROSHI Assist.Prof.

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    Grant amount:\2100000 ( Direct expense: \2100000 )

    Several cell organelles assembled from microtubules (MTs) and other proteins perform an important role in cell biology. MTs are found in a great variety of situations in cells, and may occur singly, loosely ordered into bundles, or in highly ordered arrays of a define pattern. These variations are presumably related to the different functions of MTs. In order to understand such functions, several naturally occurring compounds have been used as molecular probes. T-1 and curvularin have been found to be effective mitotic inhibitors which induced barrel-shaped spindles in sea urchin eggs due to disorder of the microtubule organising centers ( MTOCs) in centrosomes. MTs are also involved in the pronuclear fusion (PF) of sea urchin eggs. Early literature contains few reports of this process, and a better understanding of the detailed role and nature of MTs awaits the discovery of new types of specific inhibitors toward PF. We made the first systematic search for microbial products that might have PF inhibitory activity using a novel bioassay system. A Streptomyces sp. K02388 was discovered, which produced a novel gamma-pyrone, named streptopyrone with pronounced inhibitory activity toward PF. A fungus, Curvularia harveyi, was found to produce a new mitotic arrester, which was named harveynone and its structure was elucidated. This compound effectively inhibited spindle formation and arrested the first cell division of sea urchin eggs. The mode of operation of these compounds are in progress.

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  • アグロバクテリウム菌が形成するクラウンゴールを用いての二次代謝産物の探索及び生産

    Grant number:63560129  1988

    日本学術振興会  科学研究費助成事業 一般研究(C)  一般研究(C)

    河津 一儀, 神崎 浩

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    Grant amount:\1400000 ( Direct expense: \1400000 )

    1.近年、植物細胞へ外来遺伝子を導入するのに、Agrobacterium tumefaciens菌のTiプラスミドが、ベクターとして用いられているが、一般的実用化のために解決すべき問題が残っている。形質転換した植物組織よりの本細菌の除去と形質転換頻度の向上である。本研究は、これらの問題点を解決するため、本細菌感染に対する制御物質を、微生物資源に求め、その物質の化学的性状およびその生理作用を明らかにし、利用のための知見を得ることを目的としている。
    2.バクテリアの1菌株であるXanthobacter sp.KB-3001が A.tumefaciens菌を凝集させるクラウンゴール形成促進物質を生産していることを見出した。この物質PS-1は、分子量約3.5×10^6の酸性多糖で、グルコースとガラクチュロン酸とから成り、その比率は、4:1である。この物質は本菌の植物細胞への付着を助長することによりクラウンゴール形成を促進することを示した。以上の成果は、本菌を植物細胞の遺伝子組換えのベクターとして使用する際、形質転換の頻度を高め得ることを示唆するものである。
    3.A.tumefaciensに対して特異的な抗菌活性を有し、クラウンゴール形成を顕著に阻害する物質をStreptomyces sp.KBFP-2025株の培養液から単離し、Oxazolomycinと同定した。この物質KB-1はクラウンゴール形成の極めて早い段階を阻害するのみで、一旦感染が成立した植物細胞に対してはなんら阻害的な影響を示さなかった。この物質のA.tumefaciens菌及びクラウンゴール形成に対する以上の効果は、他の抗生物質には認められないものである。本物質は、100μg/mlの濃度でA.tumefaciensの増殖を抑えるが、クラウンゴールの増殖を全く阻害しなかった。以上の成果は、本物質が、クラウンゴール組織の効率よい無菌化培養に使用できることを示すものである。

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  • 放線菌における含硫アミノ酸代謝の酵素化学的研究

    Grant number:61790175  1986 - 1987

    日本学術振興会  科学研究費助成事業 奨励研究(A)  奨励研究(A)

    神崎 浩

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    Grant amount:\1090000 ( Direct expense: \1090000 )

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