文献类型：期刊文献

英文题名：Refolding, purification and characterization of an organic solvent-tolerant lipase from Serratia marcescens ECU1010

作者：Li, Suxia[1];Pang, Huaiyu[1];Lin, Kang[1];Xu, Jianhe[1];Zhao, Jian[1];Fan, Liqiang[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2011

卷号：71

期号：3-4

起止页码：171

外文期刊名：JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000292228100014)】；

基金：This work was financially supported by the National Natural Science Foundation of China (grant no. 20506037) and the Ministry of Science and Technology, PR China (grant no. 2007AA02Z225).

语种：英文

外文关键词：Lipase; Serratia marcescens; Refolding; Organic solvent tolerance; FT-IR microspectroscopy

摘要：Expression of recombinant proteins as inclusion bodies in bacteria is one of the most efficient ways to produce cloned proteins, as long as the inclusion bodies can be successfully refolded. In this study, the different parameters were investigated and optimized on the refolding of denatured lipase. The maximum lipase activity of 5000 U/L was obtained after incubation of denatured enzyme in a refolding buffer containing 20 mM Tris-HCl (pH 7.0), 1 mM Ca2+ at 20 degrees C. Then, the refolded lipase was purified to homogeneity by anion exchange chromatography. The purified refolded lipase was stable in broad ranges of temperatures and pH values, as well as in a series of water-miscible organic solvents. In addition, some water-immiscible organic solvents, such as petroleum ether and isopropyl ether, could reduce the polarity and increase the nonpolarity of the refolding system. The results of Fourier transform infrared (FT-IR) microspectroscopy were the first to confirm that lipase refolding could be further improved in the presence of organic solvents. The purified refolded lipase could enantioselectively hydrolyze trans-3-(4-methoxyphenyl) glycidic acid methyl ester [(+/-)-MPGM]. These features render the lipase attraction for biotechnological applications in the field of organic synthesis and pharmaceutical industry. Published by Elsevier B.V.