Cloning and high-level expression of the esterase gene from Klebsiella pneumoniae in Escherichia coli
2021
Chung, Y.J.
Optically active DβAcetylthioisobutyric acid (DAT) is used as a starting material in the synthesis of captopril as a valuable chiral drug in the treatment of hypertension. For an efficient DAT production by a stereoselective hydrolysis, Klebsiella pneumoniae CJ317 producing a thermostable esterase was isolated in previous study. In order to construct an esterase hyperproducing strain, the esterase gene of K. pneumoniae CJ317 was cloned into Escherichia coli and was found to show a highlevel expression, which the esterase activity of the recombinant E. coli was more than 800 times higher than that of parental strain. The nucleotide sequences of 1.6 kb BamHI EcoRI fragment containing the esterase gene were sequenced and found to include an open reading frame (ORF) of 837 bp coding 278 amino acid residues. The deduced amino acid sequence of the ORF was classified as an α/β hydrolase superfamily and showed the highest identity of 71% with that of an esterase from Pseudomonas putida MR2068. By using the recombinant E. coli as a catalyst, highly expressing the thermostable esterase gene, an efficient production of DAT by stereoselective hydrolysis with microbial cells could be useful.
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