A versatile esterase from Bacillus subtilis: cloning, expression, characterization, and its application in biocatalysis.
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.
Schmidt
Marlen
M
Institute of Biochemistry, Department of Biotechnology and Enzyme Catalysis, Greifswald University, Greifswald, Germany.
Henke
Erik
E
Heinze
Birgit
B
Kourist
Robert
R
Hidalgo
Aurelio
A
Bornscheuer
Uwe T
UT
D000438:Alcohols / Q000737:chemistry / Q000378:metabolism; D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004950:Esterases / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009154:Mutation; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013696:Temperature
D000438:Alcohols; D001426:Bacterial Proteins; D004950:Esterases; D000581:Amidohydrolases