Cloning gene encoding lysophospholipase from Bacillus halodurans CM1 to Escherichia coli DH5α

Abstract

Enzyme is a biocatalyst that is widely used in industry, for example detergent, pharmaceutical, food or oil purification. One of the most widely using enzymes for oil purification is lysophospholipase. As much as 50% of the needs of industrial enzyme are obtained from microorganisms. However, enzyme productivity of these wild type microbial strains is usually limited and cannot be applied in industry, so a genetic engineering is necessary. Cloning gene encoding for lysophospholipase was once performed in Aspergillus niger and Cryptococcus neoformans, but has never been conducted from alkalothermophilic bacteria, such as Bacillus halodurans. Bacillus halodurans CM1 is an isolate from Badan Pengkajian dan Penerapan Teknologi (BPPT). Previous research has shown that these bacteria have lipase enzymes, but the study about their properties have not been conducted. This study aims to clone the gene lysophospholipase from Bacillus halodurans CM1 to Escherichia coli DH5α using the pGEM-T easy vector. The recombinant plasmid is sequenced. The gene fragment encoding lysophospholipase was successfully obtained with size 783 base pairs and 100% similarity with gene encoding lysophospholipase from Bacillus halodurans C-125 (No access GenBank: BA000004.3).