Highly L and D enantioselective variants of horseradish peroxidase discovered by an ultrahigh-throughput selection method

被引：30

作者：

Antipov, Eugene ^{[1
]}

Cho, Art E. ^{[4
]}

Wittrup, K. Dane ^{[1
,2
]}

Klibanov, Alexander M. ^{[1
,3
]}

机构：

[1] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[2] MIT, Dept Chem Engn, Cambridge, MA 02139 USA

[3] MIT, Dept Chem, Cambridge, MA 02139 USA

[4] Korea Univ, Dept Biotechnol & Bioinformat, Seoul 136701, South Korea

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2008年 / 105卷 / 46期

基金：

美国国家卫生研究院;

关键词：

directed evolution; enzyme design; molecular modeling; redox enzymes; stereoselectivity;

D O I：

10.1073/pnas.0809851105

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

A highly efficient selection method for enhanced enzyme enantioselectivity based on yeast surface display and fluorescence-activated cell sorting (FACS) is developed and validated. its application to horseradish peroxidase has resulted in enzyme variants up to 2 orders of magnitude selective toward either substrate enantiomer at will. These marked improvements in enantioselectivity are demonstrated for the surface-bound and soluble enzymes and rationalized by computational docking studies.

引用

页码：17694 / 17699

页数：6

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