A new high-throughput screening method for determining active and enantioselective hydrolases

被引：21

作者：

Wang, Bo ^{[1
]}

Tang, Xiaoling ^{[1
]}

Ren, Gangfeng ^{[1
]}

Liu, Ji ^{[1
]}

Yu, Hongwei ^{[1
]}

机构：

[1] Zhejiang Univ, Inst Bioengn, Dept Chem & Biochem Engn, Hangzhou 310027, Zhejiang, Peoples R China

来源：

BIOCHEMICAL ENGINEERING JOURNAL | 2009年 / 46卷 / 03期

关键词：

Biocatalyst preparation; Enzyme; Microbial; Protein engineering; High-throughput screening; Fluorescein sodium salt; KINETIC RESOLUTION; HYDROLYTIC ENZYMES; ASSAY; FINGERPRINTS; EVOLUTION; REACTOR;

D O I：

10.1016/j.bej.2009.06.002

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A new high-throughput screening method using. fluorescein sodium salt as an indicator to obtain hydrolases with high enantioselectivity is developed, which is demonstrated to be sensitive and reliable. The results determined by the method correlate well with those from GC analysis. This method can be applied to determine activity and enantioselectivity of not only lipase and esterase, but also other enzymes which catalyze hydrolysis reaction releasing proton, such as the protease or amidase. Because of the application of small amount of optically pure enantiomers, screening large libraries of enzymes is allowed at low cost and in short time. (C) 2009 Elsevier B.V. All rights reserved.

引用

页码：345 / 349

页数：5

共 30 条

[1]

Badalassi F, 2000, ANGEW CHEM INT EDIT, V39, P4067, DOI 10.1002/1521-3773(20001117)39:22<4067::AID-ANIE4067>3.0.CO

[2]

2-9

[3]

Baumann M, 2001, ANGEW CHEM INT EDIT, V40, P4201, DOI 10.1002/1521-3773(20011119)40:22<4201::AID-ANIE4201>3.0.CO

[4]

2-V

[5] Single-cell high-throughput screening to identify enantioselective hydrolytic enzymes [J].

Becker, Stefan ;

Hoebenreich, Horst ;

Vogel, Andreas ;

Knorr, Janina ;

Wilhelm, Susanne ;

Rosenau, Frank ;

Jaeger, Karl-Erich ;

Reetz, Manfred T. ;

Kolmar, Harald .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2008, 47 (27) :5085-5088

[6]

BORNSCHEUER UT, 1999, ORGANIC SYNTHESIS RE

[7] Thermally responsive reversed micelles for immobilization of enzymes [J].

Chen, Hong ;

Liu, Li-Hong ;

Wang, Li-Shan ;

Ching, Chi-Bun ;

Yu, Hong-Wei ;

Yang, Yi-Yan .

ADVANCED FUNCTIONAL MATERIALS, 2008, 18 (01) :95-102

[8] SPECTROPHOTOMETRIC SUBSTRATES FOR CYTOSOLIC EPOXIDE HYDROLASE [J].

DIETZE, EC ;

KUWANO, E ;

HAMMOCK, BD .

ANALYTICAL BIOCHEMISTRY, 1994, 216 (01) :176-187

[9]

Faber K., 2000, Biotransformations in Organic Chemistry, V4th

[10] Classifying enzymes from selectivity fingerprints [J].

Grognux, J ;

Reymond, JL .

CHEMBIOCHEM, 2004, 5 (06) :826-831

← 1 2 3 →