Vol. 12, Issue 6 (2023)

Author(s):
Tejeshwari, Ningaraju TM, Lakshmipathi RN, Poornima R and Anitha Peter

Abstract:
Lipases are enzymes which catalyze the hydrolysis of triglycerides into fatty acid and glycerol. Chemical catalysts are presently used in the food and detergent industries in several processing steps in the development of many commercial products. Naturally occurring enzymes with more efficiency, energy saving and environmental friendly properties can be promising alternatives to them. The present investigation has thus been carried out with the aim for heterologous expression of lipase gene isolated from Bacillus subtilis using gene specific primers in the host E. coli. The gene consists of an open reading frame of 639 bp which encodes 213 amino acids. Initially, the isolated lipase gene was cloned in to T/A cloning vector (pTZ57 R/T) and later sub-cloned into the bacterial expression vector pET- 28a (+) and then expressed in E. coli BL21. The small scale recombinant protein expression was achieved by inducing with 1 mM IPTG at 37 °C with 4 h. of induction time. The size of the expressed recombinant protein was estimated to be 23 kDa upon electrophoresis with 12% SDS-PAGE. This lipase gene from Bacillus subtilis successfully expressed in E. coli BL21 strain can be explored for commercial application.