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

M. Schmidt, E. Henke, B. Heinze, R. Kourist, A. Hidalgo, U. T. Bornscheuer

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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.
Original languageEnglish
Pages (from-to)249-253
Number of pages5
JournalMolecular biotechnology
Volume2
Publication statusPublished - 2007

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Keywords

  • Alcohols/chemistry/metabolism Amidohydrolases/genetics/metabolism Amino Acid Sequence Bacillus subtilis/*enzymology/genetics Bacterial Proteins/genetics/*metabolism Catalysis Cloning, Molecular Enzyme Stability Esterases/genetics/*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

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