Molecular cloning of enantioselective ester hydrolase from Bacillus pumilus DBRL-191.
A gene from Bacillus pumilus expressed under its native promoter was cloned in Escherichia coli. Recombinant B. pumilus esterase (BPE) affects the kinetic resolution of racemic mixtures such as unsubstituted and substituted 1-(phenyl)ethanols (E approximately 33-103), ethyl 3-hydroxy-3-phenylpropanoate (E approximately 45-71), trans-4-fluorophenyl-3-hydroxymethyl-N-methylpiperidine (E approximately 10-13) and ethyl 2-hydroxy-4-phenylbutyrate (E approximately 7). The enzyme is composed of a 34-amino acid signal peptide and a 181-amino acid mature protein corresponding to a molecular weight of approximately 19.2kD and pI approximately 9.4. 3-D the structural model of the enzyme built by homology modelling using the atomic coordinates from the crystal structure of B. subtilis lipase (LipA) showed a compact minimal alpha/beta hydrolase fold.
Rasool
Shafaq
S
Biotechnology Division, Regional Research Laboratory, Canal Road, Jammu Tawi 180001, India.
Johri
Sarojini
S
Riyaz-ul-Hassan
Syed
S
Maqbool
Qurrat-ul-Ain
QU
Verma
Vijeshwar
V
Koul
Surrinder
S
Taneja
Subhash C
SC
Qazi
Ghulam N
GN
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D004950:Esterases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004952:Esters / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D017422:Sequence Analysis, DNA; D013237:Stereoisomerism; D013379:Substrate Specificity
D004952:Esters; D004950:Esterases