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Functional Expression of an Alkaline Lipase in Escherichia Coli Publisher



Rabbani M1 ; Mirmohammadsadeghi H1 ; Ani M2 ; Chegini KG2 ; Etemadifar Z3 ; Moazen F1
Authors
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Authors Affiliations
  1. 1. Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran
  2. 2. Department of Clinical Biochemistry and Isfahan Pharmaceutical Sciences Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran
  3. 3. Department of Biology, School of Basic Sciences, University of Isfahan, Isfahan, Iran

Source: Annals of Microbiology Published:2009


Abstract

The aim of the present study was to express and evaluate a previously cloned lipase gene. In this study, the cloned gene was subcloned in the pET15b Escherichia coli BL21(DE3) expression system. The expression of the recombinant lipase was induced using 1 mM IPTG for 3 hours. The enzyme activity was measured using p-nitrophenyl-decanoate as substrate. The recombinant lipase showed a molecular weight of 26 kDa by SDS-PAGE. Maximum activity was found at pH 9-10 and 40-50 °C. ZnCl2 at 1, 0.3, 0.1, 0.03, and 0.01 mM concentrations were found to be inhibitory to the enzyme activity and did not improve enzyme thermostability. The recombinant lipase showed an optimum temperature higher than lipase of Bacillus subtilis (its closest relative in primary structure) while similar activity in the alkaline pH.