Faculty Profiles - TAMURA Takashi

2-III-25 放線菌由来L-グルタミン酸オキシダーゼの大腸菌での大量精製系の確立(一般研究発表,日本ビタミン学会第58回大会研究発表要旨)

The crystal structure of hypothetical methyltransferase from Thermus thermophilus HB8

C Sasaki, Sugiura, I, A Ebihara, T Tamura, S Sugio, K Nagaki

PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS 64 ( 2 ) 552 - 557 2006.8

坂口智香, 水野寛士, 田村隆, 日下部均, 稲垣賢二

ビタミン 80 ( 4 ) 253 - 253 2006.4

1-II-6 抗腫瘍性酵素L-methionine γ-lyaseの活性中心におけるAsp241の機能解析(一般研究発表,日本ビタミン学会第58回大会研究発表要旨)

工藤大蔵, 岬真太郎, 瀧本明生, 田村隆, 稲垣賢二

ビタミン 80 ( 4 ) 208 - 208 2006.4

2-III-14 大腸菌の亜セレン酸同化はチオレドキシン還元酵素に依存する(一般研究発表,日本ビタミン学会第58回大会研究発表要旨)

田村隆, 高畑宗明, 阿部勝正, 三原久明, 江崎信芳, 稲垣賢二

ビタミン 80 ( 4 ) 247 - 247 2006.4

Characterization of monofunctional catalase KatA from radioresistant bacterium Deinococcus radiodurans

Kobayashi, I, T Tamura, H Sghaier, Narumi, I, S Yamaguchi, K Umeda, K Inagaki

JOURNAL OF BIOSCIENCE AND BIOENGINEERING 101 ( 4 ) 315 - 321 2006.4

Language：English Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN

Catalase plays a key role in protecting cells against toxic reactive oxygen species. Here we report on the cloning, purification and characterization of a catalase (KatA, DR1998) from the extremely radioresistant bacterium Deinococcus radiodurans. The size of purified D. radiodurans KatA monomer was 65 kDa while gel filtration revealed that the size of the enzyme was 240 kDa, suggesting that KatA formed a homotetramer in solution. Purified KatA displayed a final specific activity of 68,800 U/mg of protein. The catalase activity of KatA was inhibited by sodium azide, sodium cyanide and 3-amino-1,2,4-triazole. The absorption spectrum of KatA exhibited a Soret band at 408 nm. The position of the spectral peak remained unchanged following reduction of KatA with dithionite. No peroxidase activity was found for KatA. These results demonstrate that D. radiodurans KatA is a typical monofunctional heme-containing catalase. The stability of KatA with respect to H2O2 stress was superior to that of commercially available Aspergillus niger and bovine liver catalases. The relative abundance of KatA in cells in addition to the H2O2 resistance property may play a role in the survival strategy of D. radiodurans against oxidative damage.

DOI： 10.1263/jbb.101.315

Web of Science

Analysis of Decolorization Mechanism by Azo Dyes Decolizing Microorganism

馬場直子, 坂口博脩, 田口隆章, 早瀬伸樹, 田村隆, 稲垣賢二

岡山大学農学部学術報告 95 ( 95 ) 1 - 5 2006.2

Language：Japanese Publisher：岡山大学農学部

The mehcanism of azo dyes decolorization by Candida sp. MK-1, Aeromonas sp. B-5 and Actinobacillus sp. B-11 were analyzed. The maximal decolorization activity was observed at pH 7.5 and 30℃ on Candida sp. MK-1, at alkaline and at 35℃ on Aeromonas sp. B-5 and Actinobacillus sp. B-11. The HPLC analysis of the supernatant of the Acid Red 27 detected in the blank. The retention time of this peak matched that of a reference standard compound of 4-amino-1-naphthalenesulfonate, produced by reductive cleavage of Acid Red 27. The decolorization of azo dyes with cell free extract of Candida sp. MK-a was promoted by the addition of several coenzymes or lawsone. The remarkable promotion of decolorization was observed by the addition of glutamate dehydrogenase with α-ketoglutarate and NH4+. Therefore, it was suggested that Candida sp. MK-1 azoreductase catalyzed decolorization of azo dye by NADPH dependent reductive cleavage.

1F10-2 Purification and characterization of L-methionine γ-lyase from Streptomyces avermitilis

INAGAKI Kenji, KUDOU Daizou, HIRAI Yoshiyuki, TAMURA Takashi

18 92 - 92 2006

2-B-13 NAD依存型イソクエン酸脱水素酵素の量子酵素化学(第57回大会一般研究発表)(第57回大会研究発表要旨)

田村隆, 今田勝巳, 竹中涼, 小林一聖, 稲垣賢二

ビタミン 79 ( 4 ) 251 - 251 2005.4

化学合成独立栄養細菌Acidithiobacillus thiooxidans 由来アコニターゼの遺伝子解析と大腸菌での発現

金原陽平, 田村隆, 徳田千束, 中村淳雄, 松川寛和, 稲垣賢二

岡山大学農学部学術報告 94 1 - 7 2005

Language：Japanese Publisher：岡山大學農學部

Acidihiobacillus hiooxidans由来アコニターゼ遺伝子を大腸菌中にクローニングし、発現させ、精製及び性質検討を行った。A. hiooxidansアコニターゼはEscherichia coli JM109株で発現させ、活性が無細胞抽出液で確認された。646アミノ酸から成っており、イソクエン酸脱水素酵素遺伝子（icd）の上流に位置していた。またブタ心筋由来アコニターゼと大腸菌アコニターゼA型と高いアミノ酸配列相同性があり、ブタ心筋アコニターゼで触媒反応、基質の結合に関与するとされている27アミノ酸のうち25アミノ酸の保存が確認された。A. hiooxidansアコニターゼはDEAE-oyopearl 650M等により精製された。精製酵素は至適pH7.5、至適温度60℃であった。また40℃で1時間インキュベートを行った後にも、活性の低下は見受けられなかった。pH6.0-8.0で1時間インキュベートした後も、100％の活性を維持していた。更に本酵素は、分子量66000のモノマー酵素であることが明らかになった。なお、本論文中に記載したA. hiooxidansのアコニターゼ遺伝子を含む塩基配列はDDBJデータベースに登録されており、アクセション番号AB196983で検策できる。

ヒト肺がん細胞のセレノリン酸合成酵素Sps1,Sps2の機能解析

Other Link： http://agriknowledge.affrc.go.jp/RN/2010710742

田村隆, 山本真平, 高畑宗明, 田中英彦, 稲垣賢二

微量栄養素研究 22 147 - 152 2005

Discovery of a New Enzyme That Contains Biofactor Selenium and Characterization of Selenium Metabolism in Human Lung Adenocarcinoma Cells

TAMURA Takashi

VITAMINS 79 ( 3 ) 143 - 153 2005

Language：Japanese Publisher：THE VITAMIN SOCIETY OF JAPAN

Selenium is an essential trace element for mammals, birds, and some bacteria. Its remarkable biological effects in eukaryotes may be related to unique functions of various selenoproteins. A^<75>Se-labeled protein was purified from a human lung adenocarcinoma cell line, NCI-H441 . A homodimer of 57-kDa subunits contained selenium in the form of selenocysteine. The selenoprotein contained FAD as a prosthetic group and catalyzes NADPH-dependent reduction of insulin in the presence of thioredoxin. The subunit composition and catalytic properties were identical to those of mammalian thioredoxin reductase (TrxR). This was the first report on the biochemical relevance between selenium and Trx-mediated metabolic regulation. Interestingly, human lung adenocarcinoma cells produced the selenoenzyme TrxR over hundred times that of normal mammalian cells. A Iabile selenium donor compound monoselenophosphate, required for selenoprotein biosynthesis, is synthesized from selenide and ATP by selenophosphate synthetase (SPS).

DOI： 10.20632/vso.79.3_143

2C13-5 Large scale purification and crystal structure of L-glutamate oxidase

Other Link： http://search.jamas.or.jp/link/ui/2005136810

INAGAKI Kenji, SAKAGUCHI Chika, TAMURA Takashi, KUSAKABE Hitoshi, KASHIMA Akiko, SASAKI Chizuko, SUGIO Shigetoshi

17 109 - 109 2005

Selenophosphate synthetase genes from lung adenocarcinoma cells: Sps1 for recycling L-selenocysteine and Sps2 for selenite assimilation

T Tamura, S Yamamoto, M Takahata, H Sakaguchi, H Tanaka, TC Stadtman, K Inagaki

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 101 ( 46 ) 16162 - 16167 2004.11

Language：English Publisher：NATL ACAD SCIENCES

A labile selenium donor compound monoselenophosphate is synthesized from selenide and ATP by selenophosphate synthetase (SPS). In the present study, Sps1 and Sps2 were cloned from a cDNA library prepared from human lung adenocarcinoma cells (NCI-H441). The human lung Sps1 has been cloned as an ORF of 1,179 bp, identical in sequence to that of the recently revised human liver Sps1. The in-frame TGA codon of the lung Sps2 was genetically altered to TGT (Cys) to obtain the Sps2Cys gene. Expression of the recombinant plasmids containing Spsi or Sps2Cys was highly toxic to Escherichia coli host cells grown aerobically. Accordingly, the human lung Sps homologs were characterized by an in vivo complementation assay using a selD mutant strain. An added selenium source and a low salt concentration (0.1-0.25% NaCl) in the medium were required for reproducible and sensitive in vivo complementation. Sps2Cys effectively complemented the selD mutant, and the resulting formate dehydrogenase H activity was as high as that of WT E. coli MC4100. In contrast, only a weak complementation of the selD mutant by the Sps1 gene was observed when cells were grown in selenite media. Better complementation with added L-selenocysteine suggested involvement of a selenocysteine lyase for mobilization of selenium. Based on this apparent substrate specificity of the Sps1 and Sps2 gene products we suggest that the Sps1-encoded enzyme depends on a selenium salvage system that recycles L-selenocysteine, whereas the Sps2 enzyme can function with a selenite assimilation system.

DOI： 10.1073/pnas.0406313101

Web of Science

Saccharomyces cerevisiae 由来D-アミノ酸アセチルトランスフェラーゼの精製と性質.

2-II-26 超好熱好酸性古細菌Sulfolobus tokodaii由来シスタチオニンγ-シンターゼの精製および性質検討(第56回大会一般研究発表)

遠藤祐一, 田村隆, 倉光成紀, 田中英彦, 稲垣賢二

ビタミン 78 ( 4 ) 246 - 246 2004.4

田代(山田, 百合子, 田村隆, 田中英彦, 稲垣賢二

岡山大学農学部学術報告 93 ( 1 ) 13 - 18 2004

Language：Japanese Publisher：岡山大学農学部

Some properties of D-amino scid acetyltransferase purified from Saccharomyces cerevisiae were investigated. The enzyme was purified to homogeneity by ammonium sulfate fractionation, column chromatographies on DEAE-Toyopearl 650M, Sephacryl S-200, QAE-Toyopearl 550C and affinity chromatography with D-glutamate as a ligand. The molecular weight was estimated to be about 53,000 by gel filtration. Relative molecular mass studies indicated that the enzyme was a monomer structure. The purified enzyme had an optimum pH of 8.4 and an optimum temperature of 40C. The Km values of the purified enzyme determined with tryptophan and acetyl-CoA were 4.5 * 10 -3M, respectively. The 20 residues of N-terminal amino acid sequence were analyzed.