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

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

Institute of Molecular Biotechnology (6550)

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	249-253
Number of pages	5
Journal	Molecular Biotechnology
Volume	2
Publication status	Published - 2007

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

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.",

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author = "M. Schmidt and E. Henke and B. Heinze and R. Kourist and A. Hidalgo and Bornscheuer, {U. T.}",

note = "1860-7314 (Electronic) Journal Article",

year = "2007",

language = "English",

volume = "2",

pages = "249--253",

journal = "Molecular Biotechnology",

issn = "1073-6085",

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TY - JOUR

T1 - A versatile esterase from Bacillus subtilis: cloning, expression, characterization, and its application in biocatalysis

AU - Schmidt, M.

AU - Henke, E.

AU - Heinze, B.

AU - Kourist, R.

AU - Hidalgo, A.

AU - Bornscheuer, U. T.

N1 - 1860-7314 (Electronic) Journal Article

PY - 2007

Y1 - 2007

N2 - 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.

AB - 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.

KW - 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

M3 - Article

SN - 1073-6085

VL - 2

SP - 249

EP - 253

JO - Molecular Biotechnology

JF - Molecular Biotechnology

ER -

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

Abstract

Keywords

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