Overexpression of lipase genes in Bacillus subtilis and characterization of the recombinant lipase

China Biotechnology

2008, Vol. 28

Issue (1): 25-29 DOI:

Overexpression of lipase genes in Bacillus subtilis and characterization of the recombinant lipase

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Abstract

A Staphylococcus aureus JH　strain which can produce lipase was obtained from environment．According to polysequence comparision of prokaryote’s lipase gene published on NCBI ，we found that its sequence is very conservative． Lipase gene was obtained by PCR from genome DNA of Staphylococcus aureus JH，and then it was incorporated into plasmid pC194 and transformed into B.subtilis ．We selected the recombinant strain which is antipenicillin．The recombinant lipase was purified by ammonium sulfate fractionation and ion exchange ．SDS-PAGE was used to identify the recombinant lipase ．It was revealed that the molecular mass of the recombinant lipase is 24kDa；the recombinant lipase show maximum activity at 41℃ and pH8.0；the values of Km and Vm were found to be 0.34mM and 308 μmol•mg-1•min-1；and the metal ions Ca2+,K+and Mg2+ can activate the lipase, wheras Fe2+,Cu2+,and Co2+inhibit it.

Key words： Staphylococcus aureus lipase gene B.subtilis oveerexpression characterization

Received: 21 August 2007 Published: 25 January 2008

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Cite this article:

. Overexpression of lipase genes in Bacillus subtilis and characterization of the recombinant lipase. China Biotechnology, 2008, 28(1): 25-29.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2008/V28/I1/25

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