A versatile esterase from Bacillus subtilis: cloning, expression, characterization, and its application in biocatalysis

Marlen Schmidt; Erik Henke; Birgit Heinze; Robert Kourist; Aurelio Hidalgo; Uwe T Bornscheuer

doi:10.1002/biot.200600174

A versatile esterase from Bacillus subtilis: cloning, expression, characterization, and its application in biocatalysis

Biotechnol J. 2007 Feb;2(2):249-53. doi: 10.1002/biot.200600174.

Authors

Marlen Schmidt¹, Erik Henke, Birgit Heinze, Robert Kourist, Aurelio Hidalgo, Uwe T Bornscheuer

Affiliation

¹ Institute of Biochemistry, Department of Biotechnology and Enzyme Catalysis, Greifswald University, Greifswald, Germany.

PMID: 17136743
DOI: 10.1002/biot.200600174

Abstract

An esterase from Bacillus subtilis DSM402 (BS2) was cloned and functionally expressed in E. coli. The enzyme is active up to 50 degrees C, and the V(max) (1449 mM/min) and K(M) values (119 mM) were determined using p-nitrophenyl acetate as substrate. BS2 belongs to the few hydrolases that can act on tertiary alcohols and was therefore used to resolve racemic acetates of selected tertiary alcohols, but also to selectively remove the tert-butyl ester protecting group from peptides. In addition, the enzyme shows promiscuous amidase activity.

MeSH terms

Alcohols / chemistry
Alcohols / metabolism
Amidohydrolases / genetics
Amidohydrolases / metabolism
Amino Acid Sequence
Bacillus subtilis / enzymology*
Bacillus subtilis / genetics
Bacterial Proteins / genetics
Bacterial Proteins / metabolism*
Catalysis
Cloning, Molecular
Enzyme Stability
Esterases / genetics
Esterases / metabolism*
Gene Expression Regulation, Bacterial
Gene Expression Regulation, Enzymologic
Hydrogen-Ion Concentration
Kinetics
Molecular Sequence Data
Molecular Structure
Mutation
Sequence Homology, Amino Acid
Stereoisomerism
Temperature

Substances

Alcohols
Bacterial Proteins
Esterases
Amidohydrolases