Expression of Lipase Genes in Bacillus subtilis and Characterization of the Recombinant Lipase / 中国生物工程杂志

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,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.subtilisH11 .The recombinant lipase precipitated by(NH4)2SO4, purified by ion exchange chromatography and was identified by SDS-PAGE.It was revealed that the molecular mass of the recombinant lipase is 32kDa;the recombinant lipase show maximum activity at 41℃,pH8.0;the values of Km and Vm were found to be 0.34mmol/L and 308?mol/mg.min;The lipase can be activated by the metal ions Ca2+,K+ and Mg2+ and be inhibited by Fe2+,Cu2+,and Co2+.