Faculty Profiles - ONO Tsutomu

MISC - ONO Tsutomu

Division display >> ／ All the affair displays 1 - 74 of about 74

湿式紡糸で作るモノフィラメント状ナノファイバー Invited

小野努, 渡邉貴一

繊維学会誌 79 ( 6 ) P-188 - P-190 2023.6

高分子イオン液体材料とその分離場への期待 Invited

渡邉貴一, 小野努

分離技術 53 ( 5 ) 275 - 279 2023.5

Microfluidic Preparation of Hydrogel Microcapsules Using Aqueous Two-Phase System Formation in Monodisperse Droplets Invited

Takaichi WATANABE, Tsutomu ONO

JAPANESE JOURNAL OF MULTIPHASE FLOW 34 ( 2 ) 318 - 325 2020.6

Metal Microcapsules including liquid Invited

Tsutomu Ono, Takaichi Watanabe

C & I Commun 45 ( 3 ) 30 - 33 2020.3

マイクロ流体デバイスによる精密コロイド材料調製のための界面設計 Invited

小野努, 渡邉貴一

粉体工学会誌 55 340 - 345 2018.6

マイクロ流体デバイスを用いた単分散ポリ乳酸微粒子の連続調製 Invited

渡邉貴一, 小野努

ケミカルエンジニヤリング 63 ( 2 ) 2018.2

Interface engineering for colloid materials production using a microfluidic process

Tsutomu Ono, Takaichi Watanabe

Journal of the Society of Powder Technology, Japan 55 ( 6 ) 340 - 345 2018

Language：Japanese Publishing type：Book review, literature introduction, etc. Publisher：Society of Powder Technology

Microfluidics is a fascinating tool for colloidal materials design. It precisely controls the interface of immiscible fluids in a microchannel such as droplets, slug flow and jet flow. Monodisperse droplets prepared by microfluidic emulsification are readily converted into monodisperse particles by solvent diffusion. Furthermore, controlling the inner phase separation leads to construct the capsule structure containing oil or aqueous liquid. Based on the same concept, the rapid solidification from jet flow enables wet-spinning of nanofibers. In this review, we show our experimental procedure and the leading results and discuss key factors to develop such a microfluidic process to produce well-defined materials.

DOI： 10.4164/sptj.55.340

Study on Ultrasonic Emulsification Condition using Microchannel Device

117 ( 155 ) 37 - 40 2017.7

Effect of Pre-emulsion in Generation Nano-emulsion by Using Ultrasonic Vibration Device

33 1 - 4 2016.10

Relationship of applied voltage and droplet diameter in ultrasonic vibration device for generation of nano-emulsion

32 1 - 4 2015.10

Soap-Free Emulsion Polymerization using a Slug Flow in Microchannels

78 ( 7 ) 477 - 479 2014.7

マイクロ流路を用いた微細液滴調製に関する研究動向

渡邉貴一, 小野努

化学工学会誌 76 ( 12 ) 781 - 782 2012.12

Microfluidicsを利用したヒドロゲルカプセル新規調製法の開発

安川政宏, 小野努, 木村幸敬

化学とマイクロ・ナノシステム研究会講演要旨集 25th 2012

積層型超音波振動マイクロ流路デバイスによるエマルション生成

神田岳文, 富永宜幸, 原田拓也, 鈴森康一, 小野努, 岩渕草太郎, 伊東一行, 大河原賢一, 檜垣和孝, 吉澤雄太

精密工学会大会学術講演会講演論文集 2011 ROMBUNNO.M18 2011.3

ハイドロゲル内包マイクロカプセルの膜形成機構に及ぼす因子の検討

谷昌彦, 安川政宏, 小野努, 木村幸敬

化学工学会秋季大会研究発表講演要旨集(CD-ROM) 43rd 2011

ハイドロゲル粒子を芯物質とした鈴型構造を有する粒子の調製

谷昌彦, 安川政宏, 小野努, 木村幸敬

高分子学会予稿集(CD-ROM) 60 ( 2 Disk1 ) 2011

水相をコアテンプレートとした新規ヒドロゲルカプセル調製法の開発

安川政宏, 小野努, 木村幸敬

高分子ゲル研究討論会講演要旨集 22nd 2011

ダブルネットワーク構造を有する単分散マイクロゲル粒子の調製と機械的強度の評価

安川政宏, 小野努, 藤江史子, 岡本英子

ポリマー材料フォーラム講演予稿集 20th 2011

マイクロ流路の超音波励振によるエマルション生成

神田岳文, 原田拓也, 鈴森康一, 小野努, 岩渕草太郎, 伊東一行, 大河原賢一, 檜垣和孝

精密工学会大会学術講演会講演論文集 2010 ROMBUNNO.F26 2010.3

マイクロ流体デバイスと光重合を利用した単分散ゲル粒子調製に関する検討

安川政宏, 小野努, 木村幸敬

化学工学会年会研究発表講演要旨集(CD-ROM) 75th 2010

マイクロ流体デバイスと液・液相分離を利用した単分散ゲルカプセル調製

安川政宏, 小野努, 木村幸敬

高分子学会予稿集(CD-ROM) 59 ( 2 Disk1 ) 2010

マイクロ流路を利用した単分散複合エマルション調製における制御因子の検討

伊東一行, 安川政宏, 小野努, 木村幸敬

化学工学会年会研究発表講演要旨集(CD-ROM) 75th 2010

1P4-13 An emulsion generating device by an ultrasonic vibration and a microchannel(Poster Session)

Harada Takuya, Kanda Takefumi, Suzumori Koichi, Ono Tsutomu, Iwabuchi Sotaro, Ito Kazuyuki, Ogawara Ken-ichi, Higaki Kazutaka

Proceedings of Symposium on Ultrasonic Electronics 30 111 - 112 2009.11

光線力学的治療に有効な光増感剤内封ナノ粒子の開発

小野努, 廣田健, 木村幸敬, 白石太朗, 大河原賢一, 檜垣和孝, 阪田功

日本レーザー医学会誌 = The Journal of Japan Society for Laser Medicine 30 ( 2 ) 208 - 208 2009.7

高度に制御された巨大孔を有するマイクロカプセルの調製と応用

Eiji Kamio, Tsutomu Ono

ケミカルエンジニヤリング 54, 518-524 2009.6

水性二相系を利用したW/W/Oエマルションの調製とゲルカプセルへの応用

安川政宏, 小野努, 木村幸敬

ポリマー材料フォーラム講演予稿集 18th 2009

2Lp11 Development of high-throughput screening by culture in monodisperse gel particles

WASHIO Naoki, ONO Tsutomu, KIMURA Yukitaka

21 175 - 175 2009

水性二相系を利用したW/W/Oエマルションの調製

安川政宏, 神尾英治, 小野努

化学工学会研究発表講演要旨集 2008 ( 0 ) 643 - 643 2008

Characteristics of compacted plasmid DNA by high pressurization

Tsuyoshi Kimura, Kana Horiuchi, Kimio Kurita, Tsutomu Ono, Hidekazu Yoshizawa, Toshiya Fujisato, Akio Kishida

Nucleic Acids Symposium Series ( 51 ) 343 - 344 2007

In order to investigate the effect of pressure on the tertiary structure of plasmid DNA having the supercoiled and relaxed forms, the solution of plasmid DNA was hydrostatically pressurized at different atmosphere and 40 degrees C for various times. For dynamic light scattering (DLS) measurement of the pressurized plasmid DNA, the hydrodynamic diameters of the super-coiled and relaxed plasmid DNA were decreased with increasing pressure. Also, at constant pressure, a long period of pressure treatment effectively induced the decrease in plasmid DNA. These results suggest that the plasmid DNA was condensed by high hydrostatic pressurization. The circular dichroism (CD) spectrum of the pressurized plasmid DNA was slightly changed. For digestion by S1 nuclease, which selectively cleaves single strand DNA, the pressurized plasmid DNA was easily degraded compared to the non-pressurized plasmid DNA, suggesting that the double helix of plasmid DNA was partly dissociated to single strand by the pressure-induced compaction of plasmid DNA. These results indicate that high hydrostatic pressurization is one of powerful tools for preparing the compacted plasmid DNA.

DOI： 10.1093/nass/nrm172

Fabrication of a silica monolith micro-bioreactor entrapping highly activated lipase by a two-step sol-gel method

K. Kawakami, D. Abe, T. Urakawa, S. Sakai, T. Ono, H. Ijima

CHISA 2006 - 17th International Congress of Chemical and Process Engineering 2006.12

Micro-bioreactors are effective for high throughput production of expensive products from small amounts of substrates. Lipases are versatile enzymes for chiral syntheses, and highly activated when immobilized in alkyl-substituted silicates by the sol-gel method. For application of the immobilized lipases to a continuous-flow system, a micro-bioreactor loaded with a macroporous silica monolith is well suited, because it can easily be integrated with a separator for optical resolution. We attempted to develop a micro-capillary bioreactor containing a silica monolith-immobilized lipase. A silica monolith without shrinkage during lyophilization of the hydrogel was first formed from a 1:4 mixture of tetramethoxysilane (TMOS) and methyltrimethoxysilane (MTMS). It was then coated with silica precipitates entrapping lipase, derived from a 1:4 mixture of TMOS and n-butyltrimethoxysilane (BTMS). As a result, the monolith treated with the BTMS-based silica entrapping lipase, exhibited approximately ten times higher esterification activity than the non-treated monolith-immobilized lipase derived from the MTMS-based silica.

マイクロリアクターによるナノエマルションの調製 (特集マイクロ化学技術の実用化)

久保田潤, 小野努, 吉澤秀和

ケミカルエンジニヤリング 51 ( 11 ) 817 - 822 2006.11

Peroxidative catalytic behavior of cytochrome c solubilized in reverse micelles

Tsutomu Ono, Masahiro Goto

Biochemical Engineering Journal 28 ( 2 ) 156 - 160 2006.2

Language：English

Solubilization of horse heart cytochrome c into reverse micelles, a self-assembled nanostructure composed of sodium di-2-ethylhexyl sulfosuccinate (AOT), enhances the peroxidase activity in the presence of hydrogen peroxide. The catalytic activity of cytochrome c hosted in reverse micellar solution is 10-fold higher than that in water, and which depends on Wo, a molar ratio of water to AOT, and pH in aqueous droplets. In addition, the fluorescence intensity based on the tryptophan residue and the visible absorption identifying the axial Met 80-Fe bond imply that the reverse micellar solubilization induces the cleavage of the heme crevice and thus enables the formation of peroxidase-like peroxide-heme complexes. These results suggest that cytochrome c solubilized in reverse micelles provides a homogeneous catalyst as peroxidase available in organic media. © 2005 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.bej.2005.10.005

2L09-2 Protective effect of oligoglucosamine on the enzymatic degradation of liposome-entrapped DNA

MURAKAMI Sunao, ONO Tsutomu, SAKAI Shinji, IJIMA Hiroyuki, KAWAKAMI Koei

18 220 - 220 2006

Production of nanovesicles entrapping nucleic acids

T Ono, M Morita, S Sakai, H Ijima, K Kawakami

JOURNAL OF BIOTECHNOLOGY 118 S72 - S72 2005.8

2E10-5 Development of DNA-entrapping nano-vesicles using reverse micelles

MURAKAMI Sunao, KAWASHIMA Ayako, ONO Tsutomu, SAKAI Shinji, IJIMA Hiroyuki, KAWAKAMI Koei

17 152 - 152 2005

1H17-2 A novel approach for constructing microvascular system with desired structure

TAKEI Takayuki, SAKAI Shinji, ONO Tsutomu, IJIMA Hiroyuki, KAWAKAMI Koei

17 217 - 217 2005

Mammalian cell encapsulation in monodisperse subsieve-sized (<100 μm) capsules

S. Sakai, K. Kawabata, T. Ono, H. Ijima, K. Kawakami

Transactions - 7th World Biomaterials Congress 1510 2004.12

A method for preparing monodisperse mammalian cell-enclosing subsieve-sized microcapsules from viscous polymer solution was developed. The droplet breakup technique with an immiscible liquid coflowing stream was applied for the preparation of subsieve-sized mammalian cell-enclosing capsules. The reaction was found to enhance cell viability due to reduced resistance to the transport of oxygen, nutrients and metabolites. Mechanical strength and biocompatibility were also found to be enhanced.

DNA extraction by cationic reverse micelles

Masahiro Goto, Ayumi Momota, Tsutomu Ono

Journal of Chemical Engineering of Japan 37 ( 5 ) 662 - 668 2004.5

Language：English

Deoxyribonucleotide (DNA) was successfully extracted in a few hours by reverse micelles, which were formed and functionalized by a cationic quaternary ammonium surfactant in isooctane. Several parameters which affect the DNA transfer from an aqueous phase to a reverse micellar phase, such as the natures of a surfactant and a solvent, salt concentration in a feed solution, and hydrophobicity of micelles, were investigated. The use of a cationic surfactant was found to yield a high transfer rate of DNA into reverse micelles. We have attributed the DNA transfer to the electrostatic interaction between the cationic surfactant and the negatively charged DNA. We found that the higher the hydrophobicity of a surfactant used to form micelles, the better the DNA transfer efficiency in the forward extraction. Preferential extraction of DNA was achieved by using twin long-alkyl chains of cationic surfactants. However, pH in the aqueous feed solution did not affect the extraction performance of reverse micelles. On the other hand, alcohol, as a co-solvent plays an important role in the back extraction of DNAs. Complete back-extraction of DNAs was achieved without any conformational change by adding an appropriate alcohol to the reverse micellar phase. © 2004 The Society of Chemical Engineers, Japan.

DOI： 10.1252/jcej.37.662

2F15-3 Effect of cell-suspension viscosity on subsieve-sized capsule diameter

SAKAI Shinji, KAWABATA Kenji, ONO Tsutomu, IJIMA Hiroyuki, KAWAKAMI Koei

16 187 - 187 2004

3I09-4 Preparation of chitosan oligosaccharide-liposome complexes utilizing freeze-thawing method

MURAKAMI Sunao, SAKAI Shinji, ONO Tsutomu, IJIMA Hiroyuki, KAWAKAMI Koei

16 228 - 228 2004

Protein refolding membrane reactor using reverse micelles

Ono Tsutomu, Momozaki Tatsuro, Ijima Hiroyuki, Kawakami Koei

15 165 - 165 2003

Development of microcapsule for single cell encapsulation utilizing co-flowing liquid-liquid stream

Kawabata Kenji, Sakai Sinji, Ono Tsutomu, Ijima Hiroyuki, Kawakami Koei

15 91 - 91 2003

Optimization of co-culture conditions of hepatocytes and bone marrow cells for expression and maintenance of liver specific functions

Ijima Hiroyuki, Matsuo Takeshi, Murakami Sunao, Takei Takayuki, Sakai Shinji, Ono Tsutomu, Kawakami Koei

15 89 - 89 2003

ナノ空間で起こる逆ミセルの生化学反応とその応用

バイオサイエンスとインダストリー 60 ( 11 ) 747 - 750 2002

1161 Effect of chemical composition of alginate on properties of aminopropyl-silicate microcapsule for bioartificial pancreas

Sakai Shinji, Ono Tsutomu, Ijima Hiroyuki, Kawakami Koei

14 214 - 214 2002

Catalytic oxidation of o-phenylenediamine by cytochrome c encapsulated in reversed micelles

T Ono, K Kawakami, M Goto, S Furusaki

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC 11 ( 4-6 ) 955 - 959 2001.1

Language：English Publisher：ELSEVIER SCIENCE BV

Cytochrome c solubilized in reversed micelles formulated with di-2-ethylhexyl sulfosuccinate (AOT) has allowed to catalyze the oxidation of o-phenylenediamine (o-PDA) with hydrogen peroxide. The oxidative reaction did not occur effectively by cytochrome c in water. The nanostructural environment in reversed micelles seemed to perturb the three-dimensional structure of the active site of cytochrome c. The product obtained in reversed micelles had a specific absorption spectrum with the absorption peak at 720 nm, indicating a blue-colored solution, whereas common peroxidases such as horseradish peroxidase provided a yellow-colored product in water with the absorption peak at 420 nm. The difference in the oxidative products suggests that AOT molecules play an important role in the specificity of o-PDA oxidation. The dependency of the product species on the pH of the inner droplet in reversed micelles indicated that the specific oxidative product was prepared by electrostatic interaction with AOT molecules. (C) 2001 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S1381-1177(00)00173-9

Selective Extraction of Active α-Chymotrypsin by Reversed Micelles

Masahiro Goto, Maki Korenaga, Tsutomu Ono, Shintaro Furusaki

Kagaku Kogaku Ronbunshu 27 ( 2 ) 181 - 185 2001

Language：English

Extraction behavior of active and denatured α-chymotrypsin is investigated by the reversed micellar extraction technique. The anionic surfactant dioleyl phosphoric acid (DOLPA) selectively extracted the active protein over the denatured protein. However, the reversed micelles using AOT do not afford selectivity. Based on the difference in the structural properties between the active and the heated α-chymotrypsin, the extraction behavior by the DOLPA reversed micelles is discussed. Key words: extraction, reverse micelle, surfactant, protein separation, AOT. © 2001, The Society of Chemical Engineers, Japan. All rights reserved.

DOI： 10.1252/kakoronbunshu.27.181

生体分子とナノ集合体の融合による新機能発現

化学工業 52 ( 3 ) 192 - 197 2001

Behavior of protein refolding in reversed micellar solution.

Ono Tsutomu, Ando Takashi, Nagao Tomoaki, Iijima Hiroyuki, Kawakami Koei

13 201 - 201 2001

Fabrication and evaluation of aminopropylsilicate-titanate membrane toward microcapsule-shaped bioartificial organs

Sakai Shinji, Ono Tsutomu, Iijima Hiroyuki, Kawakami Koei

13 201 - 201 2001

Important parameters affecting efficiency of protein refolding by reversed micelles

M Goto, Y Hashimoto, T Fujita, T Ono, S Furusaki

BIOTECHNOLOGY PROGRESS 16 ( 6 ) 1079 - 1085 2000.11

Language：English Publisher：AMER CHEMICAL SOC

Refolding of denatured RNase A as a model of inclusion bodies was performed by reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) in isooctane. In the novel refolding process, a solid-liquid extraction was utilized as an alternative to the ordinary protein extraction by reversed micelles based on a liquid-liquid extraction. First, the effects of operational parameters such as concentration of AOT, W-o (= [H2O]/[AOT]), and pH were examined on the solubilization of solid denatured proteins into a reversed micellar solution. The solubilization was facilitated by a high AOT concentration, a high W-o value, and a high pH in water pools. These conditions are favorable for the dispersion of the solid protein aggregates in an organic solvent. Second, the renaturation of the denatured RNase A solubilized into the reversed micellar solution was conducted by addition of glutathione as a redox reagent. A complete renaturation of RNase A was accomplished by adjusting the composition of the redox reagent even at a high protein concentration in which protein aggregation would usually occur in aqueous media. In addition, the renaturation rates were improved by optimizing water content (W-o) and the pH of water pools in reversed micelles. Finally, the recovery of renatured RNase A from the reversed micellar solution was performed by adding a polar organic solvent such as acetone into the reversed micellar solution. This precipitation method was effective for recovering proteins from reversed micellar media without any significant reduction in enzymatic activity.

DOI： 10.1021/bp000073m

Catalytic capabilities of lipase immobilized into organically modified silicates by sol-gel method

SY Furukawa, T Ono, H Ijima, K Kawakami

KAGAKU KOGAKU RONBUNSHU 26 ( 6 ) 882 - 886 2000.11

Language：Japanese Publisher：SOC CHEMICAL ENG JAPAN

The capabilities of Candida rugosa lipase, which was immobilized into organically modified silicates formed on kieselguhr such as Celite 545 are investigated in esterification of (-)- or (+)-menthol with butyric acid. In this study, we optimized the preparation conditions such as a molar ratio of precursors. From these investigations, we find that the optimized hybrid gel-immobilized lipase is derived from a mixture of dimethyldimethoxysilane and tetramethoxysilane at a molar ratio of 4 : 1 and its enantioselectivity, which is defined as a ratio of the initial reaction rate of (-)-menthol to that of (+) -menthol, is enhanced up to 41 at 75 degreesC. In addition, we observe the temperature dependence of these immobilized lipases. Consequently, the optimized hybrid gel-immobilized lipase shows higher activity than deposited lipase on Celite at all temperatures, and it is suggested that this activation results from a remarkable enhancement of the V-max value.

DOI： 10.1252/kakoronbunshu.26.882

Control of molecular weight cut-off for immunoisolation by multilayering glycol chitosan-alginate polyion complex on alginate-based microcapsules

S Sakai, T Ono, H Ijima, K Kawakami

JOURNAL OF MICROENCAPSULATION 17 ( 6 ) 691 - 699 2000.11

Language：English Publisher：TAYLOR & FRANCIS LTD

Glycol chitosan is a positively charged polysaccharide which is water-soluble at pH 7.4, and is able to form a polyion complex (PIC) with anionic polymers, such as alginate. The authors attempt to develop a novel type of alginate-based microcapsule using this glycol chitosan for a islets-encapsulated bioartificial pancreas. The number of layers composed of glycol chitosan-alginate (GC-Alg) PIC were optimized, in order to cut off immunoglobulin transport and to protect encapsulated islets from the host immune reaction, and the transport characteristics were evaluated of glucose, bovine serum albumin (BSA) and gamma-globulin. To add mechanical stability to the microcapsule, calcium ions, which crosslinked the alginate polymers close to the interface between core Ca-alginate and multilayered membrane, were partially substituted with barium ions after the formation of multilayered Ca-alginate gel beads. The partition coefficients of BSA and gamma-globulin were decreased with the increasing number of layers. The immunoisolation was achieved against gamma-globulin with four layers of the GC-Alg PIC membrane, while BSA could permate the membrane. The foul-layered Ba-alginate gel bead had a good permeability for glucose, giving a diffusion coefficient corresponding to 80% of that in pure water. Insulin secretion from the islets in the four-layered Ba-alginate microcapsule was satisfactorily observed with the fractional stimulation ratio of 2.17. This result indicates that the encapsulated islets maintained their viability even after encapsulation. It was, thus, shown that the Ba-alginate microcapsule with four layers of the GC-Alg PIC membrane is promising as the microencapsulation material for a bioartificial pancreas.

DOI： 10.1080/02652040050161684

Selective Separation of Biomolecules by Reversed Micelles : Extraction of Proteins and Nucleic Acids

GOTO Masahiro, ONO Tsutomu, FURUSAKI Shintaro

MEMBRANE 25 ( 1 ) 23 - 31 2000

Language：Japanese Publisher：THE MEMBRANE SOCIETY OF JAPAN

Reversed micelles, which are nanostructural molecular assemblies formed by surfactant molecules in an organic solvent, are a promising separation tool for biomolecules such as amino acids, proteins and nucleic acids. In the separation by reversed micelles, biomolecules transfer from an aqueous solution into a reversed micellar phase through the interaction between surfactant molecules and a target biomolecule : e.g. electrostatic interaction, hydrophobic interaction, hydrogen bonding and/or affinity interaction. In particular, the electrostatic interaction has been found to play an important role in the degree of extraction, and the design of reversed micelles often decides the success of separation efficiency. In this article, we reviewed the recent investigation concerning the reversed micellar extraction techniques for biomolecules ; mainly proteins and deoxyribonucleic acids (DNAs). We also describe the transfer mechanism of proteins through the protein-surfactant complexes at the liquid-liquid interface. DNAs were first demonstrated to be transferred from an aqueous solution to the reversed micellar phase by using a cationic surfactant. Furthermore, we introduce recent novel application of reversed micelles to the separation of biomolecules.

DOI： 10.5360/membrane.25.23

Other Link： https://jlc.jst.go.jp/DN/JALC/00087896763?from=CiNii

Reversed micelles as novel protein refolding media

ACS Symp. Series 740 374 - 383 2000

ゾルーゲル法により調製した有機-無機ハイブリッドシリカ固定化リパーゼの触媒性能

化学工学論文集 26 ( 6 ) 882 - 886 2000

機能性流体逆ミセルのバイオ分野への応用

ケミカルエンジニアリング 54 ( 6 ) 418 - 424 2000

逆ミセルの生物工学的応用

小野努, 後藤雅宏

表面 37 ( 1 ) 59 - 66 1999

Extraction of DNA by reversed micelles

Masahiro Goto, Tsutomu Ono, Akihiko Horiuchi, Shintaro Furusaki

Journal of Chemical Engineering of Japan 32 ( 1 ) 123 - 125 1999

Language：English Publisher：Society of Chemical Engineers, Japan

Deoxyribonucleotide (DNA) from salmon testes was successfully extracted by reversed micelles that are formed by a cationic surfactant in isooctane. The degree of extraction of DNA was strongly influenced by the type of surfactant in the organic solvent and ionic strength in the material aqueous phase. The driving force of the DNA transfer was electrostatic interaction between cationic surfactants and negatively charged DNA. Cationic surfactants possessing two long-alkyl chains are one of the best candidates for the successful transfer of DNA from an aqueous solution to a reversed micellar solution. Copyright © 1999 The Society of Chemical Engineers,.

DOI： 10.1252/jcej.32.123

Peroxidase activity of cytochrome c in organic media

Ono Tsutomu, Kawakami Koei, Goto Masahiro, Furusaki Shintaro

11 252 - 252 1999

Factors affecting protein transfer into surfactant-isooctane solution: A case study of extraction behavior of chemically modified cytochrome c

T Ono, M Goto

BIOTECHNOLOGY PROGRESS 14 ( 6 ) 903 - 908 1998.11

Language：English Publisher：AMER CHEMICAL SOC

The extraction mechanism of proteins by surfactant molecules in an organic solvent has been investigated using a chemically modified protein. We conducted guanidylation on lysine residues of cytochrome c by replacing their amino groups with homoarginine to enhance the protein-surfactant interaction. Results have shown that guanidylated cytochrome c readily forms a hydrophobic complex with dioleyl phosphoric acid (DOLPA) through hydrogen bonding between the phosphate moiety and the guanidinium groups. Although improved protein-surfactant interaction activated the formation of a hydrophobic complex at the interface, it could not improve the protein transfer in isooctane. It has been established that the protein extraction mechanism using surfactant molecules is mainly governed by two processes: formation of an interfacial complex at the oil-water interface and the subsequent solubilization of the complex into the organic phase. In addition, a kinetic study demonstrated that guanidylation of lysine accelerated the initial extraction rate of cytochrome c. This fact implies that the protein transferability from aqueous phase into organic phase depends on the protein-surfactant interaction which can be modified by protein surface engineering.

DOI： 10.1021/bp9800844

Extraction and activity of chymotrypsin using AOT-DOLPA mixed reversed micellar systems

Masahiro Goto, Yoshio Ishikawa, Tsutomu Ono, Fumiyuki Nakashio, T. Alan Hatton

Biotechnology Progress 14 ( 5 ) 729 - 734 1998.9

Language：English Publisher：AIChE

Novel reversed micellar solutions formulated with a mixture of AOT (dioctyl sulfosuccinate) and DOLPA (dioleyl phosphoric acid) show good potential for use in reversed micellar protein extraction operations. Chymotrypsin is easily extracted from an aqueous phase into organic isooctane containing 10 mM AOT and DOLPA in a 4:1 ratio. The extraction ability of the mixed reversed micelles of 10 mM was higher than that of 200 mM AOT alone. The results of extraction indicated that the AOT-DOLPA mixed reversed micelles are very useful for separating and enriching chymotrypsin. Back-extraction of chymotrypsin from the organic phase to a fresh aqueous phase is also accomplished by adding an alcohol to the organic phase. Although the backtransfer of chymotrypsin from the reversed micelles formed by AOT alone is very slow and difficult, in the AOT-DOLPA mixed reversed micelles, the back-extraction can be achieved completely by addition of 10% (v/v) isobutyl alcohol to the reversed micellar phase. The time to attain to the equilibrium of back-extraction was reduced from more than 24 to 2 h by adding the alcohol. On the basis of the activity data, the best composition of AOT and DOLPA was a 4:1 ratio and the total surfactant concentration was 10 mM. The activity of chymotrypsin recovered from the mixed reversed micelles was higher than that of the initial protein before the forward-transfer. This result means that the novel mixed reversed micellar solutions are useful not only in separation but also in purification of proteins.

DOI： 10.1021/bp9800790

Protein refolding by reversed micelles utilizing solid-liquid extraction technique

Y Hashimoto, T Ono, M Goto, TA Hatton

BIOTECHNOLOGY AND BIOENGINEERING 57 ( 5 ) 620 - 623 1998.3

Language：English Publisher：JOHN WILEY & SONS INC

This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by a solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates. (C) 1998 John Wiley & Sons, Inc.

DOI： 10.1002/(SICI)1097-0290(19980305)57:5<620::AID-BIT14>3.0.CO;2-F

Catalytic properties of novel reversed micellar system on trans-esterification by α-chymotrypsin in organic media

Ly Iskandar, Tsutomu Ono, Noriho Kamiya, Masahiro Goto, Fumiyuki Nakashio, Shintaro Furusaki

Biochemical Engineering Journal 2 ( 1 ) 29 - 33 1998

Language：English Publisher：Elsevier

Novel reversed micelles formulated by sodium dioleylphosphate (SDOLP) has been developed for enzymatic reaction in organic media. Comparison study between sodium bis-(2-ethylhexyl)-sulfosuccinate (Aerosol-OT or AOT) and SDOLP, which was carried out by investigating the reactivity of each reversed micellar system as reaction media for enzymatic transesterification reaction of N-acetyl-1-phenylalanine ethyl ester (APEE) and 1-propanol, leads to the upshot that α-chymotrypsin immobilized within SDOLP reversed micelles can persevere its high catalytic activity in isooctane. The initial reaction rate of the SDOLP reversed micellar system is almost 60 times higher than that in the AOT system. Additional kinetic results, including the Michaelis constant and the maximum reaction rate, reveal that the inner core of the SDOLP reversed micelles provides a better environment for enzymatic reaction than that of the conventional AOT reversed micelles. © 1998 Elsevier Science S.A. All rights reserved.

DOI： 10.1016/S1369-703X(98)00013-8

Application of reversed micelles in bioengineering

T Ono, M Goto

CURRENT OPINION IN COLLOID & INTERFACE SCIENCE 2 ( 4 ) 397 - 401 1997.8

Language：English Publishing type：Book review, literature introduction, etc. Publisher：CURRENT SCIENCE LTD

Reverse micelles have found wider biological applications recently in terms of providing an enabling hydrophilic environment for solubilized functional biomaterials in organic media. Further, separation and purification of target proteins from aqueous solution using micellar aggregates created in hydrophobic media as well as efficient refolding of proteins in micellar solutions have become possible.

Design of surfactants suitable for protein extraction by reversed micelles

Masahiro Goto, Tsutomu Ono, Fumiyuki Nakashio, T. Alan Hatton

Biotechnology and Bioengineering 54 ( 1 ) 26 - 32 1997.4

Language：English Publisher：John Wiley & Sons Inc

New surfactants have been synthesized for potential use in reversed micellar protein extraction operations. Preferential solubility of the surfactant in an aliphatic solvent such as hexane, heptane, or isooctane and the formation of reversed micelles accompanied with solubilization of significant quantities of water can be achieved by using strongly hydrophobic, twin alkyl chains as the hydrophobic moiety. Different surfactants having identical water-solubilizing capacities can have significantly different behavior in protein extractions, where extraction efficiency appears to be governed by the nature of the interfacial complex that forms between surfactants and proteins. Bulky surfactant chains provide a steric hindrance to the adsorption of the surfactant to the protein surface, thus inhibiting solvation of the protein/surfactant complex, and hence protein extraction. Under these conditions, a precipitate forms either in the bulk aqueous phase or at the interface. Surfactants that can form a close-packed complex with the protein are excellent protein-solubilizing agents. Dioleyl phosphoric acid (DOLPA) appears to be the best surfactant currently available for protein extraction.

DOI： 10.1002/(SICI)1097-0290(19970405)54:1<26::AID-BIT3>3.0.CO;2-#

分子集合体を利用した抽出分離技術 (特集分離技術シンポジウム)

後藤雅宏, 小野努

分離技術 27 ( 3 ) 193 - 198 1997.4

Effect of organic solvents on extraction behaviour of cytochrome c using reversed micellar solutions

Tsutomu Ono, Takayuki Miyazaki, Masahiro Goto, Fumiyuki Nakashio

Solvent Extraction Research and Development 1996 1 - 9 1996.12

The effect of organic solvents on the extraction behaviour of cytochrome c has been investigated with reversed micellar solutions. An aliphatic solvent such as hexane, octane or isooctane was suitable for protein extraction operations using the reversed micelles. Organic solvents having a different hydrophobicity show significantly different behaviour in protein extraction, where extraction efficiency appears to be governed by the nature of the interfacial complex that is formed by surfactants and proteins. Solvent effects on the protein extraction are more dominant in the AOT system rather than that in the DOLPA system. Addition of alcohol appears to decrease the activity of proteins in the back-extraction step, although it is effective for recovering proteins from the reversed micellar solutions.

Extraction behavior of hemoglobin using reversed micelles by dioleyl phosphoric acid

T Ono, M Goto, F Nakashio, TA Hatton

BIOTECHNOLOGY PROGRESS 12 ( 6 ) 793 - 800 1996.11

Language：English Publisher：AMER CHEMICAL SOC

A new surfactant, dioleyl phosphoric acid (DOLPA), has been applied to the extraction of hemoglobin using reversed micelles. The reversed micelles formed by DOLPA can easily extract hemoglobin from aqueous to reversed micellar solutions. DOLPA is the first surfactant to extract hemoglobin completely without using any cosurfactants. On the basis of the difference between DOLPA and AOT reversed micelles in the forward extraction behavior of hemoglobin, the nature of the interfacial complex that would be formed between surfactants and hemoglobin at the oil-water interface was found to be the dominant factor in determining the extraction efficiency of hemoglobin by reversed micelles. In addition, back-transfer studies of hemoglobin from the DOLPA reversed micelles were also carried out by the phase transfer method. It was found that hemoglobin, once dissolved into the DOLPA reversed micelles, is not transferred to a fresh aqueous solution even when the conditions are adjusted to not allow the forward transfer of hemoglobin. However, the addition of several kinds of alcohol drastically improved the yield in the back-transfer of hemoglobin. The efficiency in the back-transfer of hemoglobin strongly depends on the aqueous conditions that are in contact with the reversed micelles, such as pH, ionic strength, and alcohol concentration. A pH higher than the pi of hemoglobin, a salt concentration lower than that of the water pool, and the proper concentration of alcohol are required for the recovery aqueous phase to ensure the back-transfer of hemoglobin from the DOLPA reversed micelles.

DOI： 10.1021/bp960055y

Novel immobilized enzymes utilizing reversed micellar solutions as an optical resolution tool

T Ono, M Goto, F Nakashio, TA Hatton

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN 29 ( 4 ) 695 - 697 1996.8

Reversed micelles recognize an active protein

Masahiro Goto, Tsutomu Ono, Fumiyuki Nakashio, T. Alan Hatton

Biotechnology Techniques 10 ( 3 ) 141 - 144 1996

Language：English Publisher：Springer Netherlands

The extraction behavior of native and heated-denatured α-chymotrypsin has been investigated with two different reversed micellar systems. A large difference in the degree of extraction was observed for the native relative to the denatured α-chymotrypsin. In particular, mixed reversed micelles formulated with DOLPA (dioleyl phosphoric acid) and AOT show a high selectivity for the active α-chymotrypsin.

DOI： 10.1007/BF00158935

Protein Extraction by New Reversed Micelles with Di(Tridecyl) Phosphoric Acid

Masahiro Goto, Motohiro Kuroki, Tsutomu Ono, Fumiy uki Nakashio

Separation Science and Technology 30 ( 1 ) 89 - 99 1995

Language：English

Di(tridecyl) phosphoric acid (DTDPA) was synthesized as a new surfactant for protein extraction by reversed micelles. DTDPA can form stable reversed micelles in the concentration range above 1 mM. The size of the DTDPA reversed micelles was smaller than that of conventional AOT (sodium-di-2-ethylhexyl sulfosucci-nate) reversed micelles under the same conditions. The formation of reversed micelles using DTDPA can be controlled by adjusting the aqueous pH. The extraction behavior of some proteins from an aqueous phase into the reversed micellar phase of DTDPA has been investigated by varying the experimental conditions (pH, ionic strength, surfactant concentration). It was found that reversed micelles with DTDPA can extract many proteins. In particular, hemoglobin, which is difficult to extract with an AOT reversed micelle, can be extracted in the DTDPA reversed micelles. In the new reversed micellar system, phase separation was very fast even at a high concentration of DTDPA. © 1995, Taylor &amp
Francis Group, LLC. All rights reserved.

DOI： 10.1080/01496399508012215

有機媒体中における分子集合系のタンパク質の分離場としての利用--逆相ミセルによるタンパク質の抽出とその可溶化機構

小野努, 後藤雅宏

表面 32 ( 9 ) p601 - 611 1994.9

Protein extraction and extraction mechanism by reversed micelles

32 ( 9 ) 601 1994

　 PREV - NEXT 　

1