Cloning, Expression and Characterization of a Lipase Gene from Marine BacteriumPseudoalter

Cloning, Expression and Characterization of a

Lipase Gene from Marine

BacteriumPseudoalteromonas lipolytica SCSIO

04301

SU Hongfei;MAI Zhimao;ZHANG Si

【期刊名称】《中国海洋大学学报（英文版）》 【年(卷),期】2016(015)006

【摘要】A lipase gene, lip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80 kDa. Lip1233 was grouped into the lipase family X because it contained a highly conserved motif GHSLG. The recombinant enzyme was purified with Ni-NTA affinity chromatography. The optimal temperature and pH value of Lip1233 were 45℃ and 8.0, respectively. It retained more than 70% of original activity after being incubated in pH ranging from 6.0 to 9.5 for 30 min. It was stable when the temperature was below 45℃, but was unstable when the temperature was above 55℃. Most metal ions tested had no significant effect on the activity of Lip1233. Lip1233 remained more than original activity in some organic solvents at the concentration of 30% (v/v). It retained more than 30% activity after incubated in pure organic solvents for 12 h, while in hexane the activity was nearly 100%. Additionally, Lip1233 exhibited