Isolation and identification of high-yield lipase producing bacteria and analysis of lipase gene

Objective:To separate natural bacteria producing high activities lipase from environment,and clone its lipase gene.Methods:High activities lipase producing bacteria was screened from soil which contained plenty of oils and fats.After identifying bacteria,PCR amplified lipase gene which was cloned to pMD18-T Vector.The recombint plasmid was transformed into E.Coli DH5α to build the engineering bacteria.At last,the lipase gene was identified with two restriction endonuclease digesting and sequencing.Results: After identifiying,4 producing high activitie lipases bacteria strains were obtained,one of which was a Staphylococcus aureus producing highest activities lipase,its lipase gene was amplified by PCR,then separated and identified as a new mutant lipase gene by restriction endonuclease digesting and sequencing.standard lipase gene registered on NCBI(EU310372),The secondary structure of the new mutant lipase gene was more different from standard lipase.Conclusion:A strain of high activities lipase producing bacteria-Staphylococcus aureus is successfully isolated and the cloned lipase gene is a new discovered mutant type.