Rational Protein Design of Paenibacillus barcinonensis Esterase EstA for Kinetic Resolution of Tertiary Alcohols

被引：28

作者：

Bassegoda, Arnau ^{[1
,2
]}

Nguyen, Giang-Son ^{[1
]}

Schmidt, Marlen ^{[1
]}

Kourist, Robert ^{[1
]}

Diaz, Pilar ^{[2
]}

Bornscheuer, Uwe T. ^{[1
]}

机构：

[1] Ernst Moritz Arndt Univ Greifswald, Dept Biotechnol & Enzyme Catalysis, Inst Biochem, D-17487 Greifswald, Germany

[2] Univ Barcelona, Dept Microbiol, E-08028 Barcelona, Spain

来源：

CHEMCATCHEM | 2010年 / 2卷 / 08期

关键词：

alcohols; enzyme catalysis; esterases; kinetic resolution; protein engineering; DIRECTED EVOLUTION; BACILLUS-SUBTILIS; ENANTIOSELECTIVITY; INHIBITORS; CARBOXYLESTERASES; IDENTIFICATION; ENZYMES;

D O I：

10.1002/cctc.201000053

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

Protein engineering is a very powerful tool to optimize enzymes for specific applications and thus provide important chiral building blocks such as tertiary alcohols. By use of structural comparisons, esterase from Paenibacillus barcinonensis (EstA) was engineered to convert tertiary alcohol esters with excellent enantioselectivity. Whereas the wild-type enzyme converts 1,1,1-trifluoro-2-phenylbut-3-yn-2-yl acetate with very low activity and enantioselectivity (E = 12, at 4 degrees C), several mutants show a significantly increased enantioselectivity, for example E > 100 for mutant EstA-AGA, under the same reaction conditions. Furthermore, the range of tertiary alcohols obtained in enantiopure form was also broadened for EstA mutants.

引用

页码：962 / 967

页数：6

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