Gene Cloning. Expression and Characterization of the Lipase from Bacillus pumilus S6

Abstract

Abstract: A lipase-producing strain was screened from the soil samples. It was identified as Bacillus pumilus by the 16S rRNA method. According to the reported lipase genes from Bacillus, the primers were derived and the full-length gene was cloned, named “lipS6”, which had the size of 648 bp, encoding 215 amino acids. “lipS6” held 96% homology with the reported lipase gene B26. Recombinant plasmid pET32a-lipS6 was construced and expressed in E. coli. “lipS6”could be expressed in soluble form, and the lipase activity reached 2 000 U/L. The optimum temperature of recombinant lipase LipS6 was 30℃, and the optimum pH was 9. Low concentrations of Ca²⁺ and Mn²⁺ could activate the activity of lipase LipS6. The harmful effect of hydrophobic organic solvent on the lipase LipS6 was little. LipS6 could catalyze the esterification reaction in the n-hexane system, with the myristic acid and n-butanol as the optimum substrates. These properties show that LipS6 can be used in the fields of pulp and paper, leather, textiles and bioenergy.

Key words: Lipase, Bacillus pumilus, Gene cloning, Recombinant expression, Characterization

Su Erzheng, Wu Xiangping, Gao Bei, Wei Dongzhi,. Gene Cloning. Expression and Characterization of the Lipase from Bacillus pumilus S6[J]. Biotechnology Bulletin, 2014, 0(4): 132-138.

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