Open AccessMolecular cloning of enantioselective ester hydrolase from Bacillus pumilus DBRL‐191
Author(s) -
Rasool Shafaq,
Johri Sarojini,
RiyazulHassan Syed,
Maqbool QurratulAin,
Verma Vijeshwar,
Koul Surrinder,
Taneja Subhash C.,
Qazi Ghulam N.
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.06.022
Subject(s) - bacillus pumilus , hydrolase , escherichia coli , bacillus subtilis , stereochemistry , enantioselective synthesis , molecular cloning , chemistry , amino acid , esterase , biochemistry , enzyme , peptide sequence , biology , gene , bacteria , catalysis , genetics
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 ? 33 –103), ethyl 3‐hydroxy‐3‐phenylpropanoate ( E ? 45 –71), trans ‐4‐fluorophenyl‐3‐hydroxymethyl‐ N ‐methylpiperidine ( E ? 10 –13) and ethyl 2‐hydroxy‐4‐phenylbutyrate ( E ? 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 ?19.2 kD and pI ? 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 α/β hydrolase fold.