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Rational Protein Design of Paenibacillus barcinonensis Esterase EstA for Kinetic Resolution of Tertiary Alcohols
Author(s) -
Bassegoda Arnau,
Nguyen GiangSon,
Schmidt Marlen,
Kourist Robert,
Diaz Pilar,
Bornscheuer Uwe T.
Publication year - 2010
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201000053
Subject(s) - enantiopure drug , kinetic resolution , esterase , protein tertiary structure , chemistry , protein engineering , mutant , rational design , tertiary alcohols , biocatalysis , enzyme , directed evolution , alcohol , enantioselective synthesis , stereochemistry , combinatorial chemistry , organic chemistry , catalysis , biochemistry , materials science , nanotechnology , reaction mechanism , gene
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 °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.