The role of the GGGX motif in determining the activity and enantioselectivity of pig liver esterase towards tertiary alcohols

被引：15
作者：
Gall, Markus [1 ]
Kourist, Robert [1 ]
Schmidt, Marlen [1 ]
Bornscheuer, Uwe T. [1 ]
机构：
[1] Ernst Moritz Arndt Univ Greifswald, Inst Biochem, Dept Biotechnol & Enzyme Catalysis, D-17487 Greifswald, Germany
来源：
BIOCATALYSIS AND BIOTRANSFORMATION | 2010年 / 28卷 / 03期
关键词：
Kinetic resolution; enantioselectivity; pig liver esterase; tertiary alcohol; molecular modeling; GGG(A) X motif; FUNCTIONAL EXPRESSION; KINETIC RESOLUTION; BACILLUS-SUBTILIS; PICHIA-PASTORIS; IDENTIFICATION; MUTANTS; CLONING; VERSATILE;
D O I：
10.3109/10242421003753803
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The GGG(A) X motif has been reported to determine the activity of esterases or lipases towards tertiary alcohols. The gamma-isoenzyme of pig liver esterase (gamma-PLE, PLE 1) contains a GGGX motif; this motif was systematically mutated to the corresponding AGGX, GAGX, GGAX, AAAX, AAGX, AGAX and GAAX motifs to investigate its influence on the activity and especially enantioselectivity in the kinetic resolution of three acetates of tertiary alcohols. None of the mutants showed higher enantioselectivity than the starting enzyme (E = 17), and mutation of the central glycine to alanine completely destroyed activity and selectivity. Hence, only subtle changes of this crucial motif lead to substantial changes in esterase properties. These findings were analyzed and interpreted by computer modeling.
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收藏
页码：201 / 208
页数：8
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