title: Production, characterization, and application of an organic solvent-tolerant lipase present in active inclusion bodies.
abstract: An organic solvent-tolerant lipase from Serratia marcescens ECU1010 (rSML) was overproduced in Escherichia coli in an insoluble form. High concentrations of both biomass (50 g cell wet weight/L culture broth) and inclusion bodies (10.5 g/L) were obtained by applying a high-cell-density cultivation procedure. Activity assays indicated that the enzymatic activity of rSML reached 600 U/L. After treatment with isopropyl ether for 12 h, the maximum lipase activity reached 6,000 U/L. Scanning electron microscopy and Fourier transform infrared microspectroscopy revealed the activation mechanism of rSML in the presence of organic solvents. rSML was stable in broad ranges of temperatures and pH values, as well as in a series of organic solvents. Besides, rSML showed the best enantioselectivity for the kinetic resolution of (±)-trans-3-(4-methoxyphenyl)glycidic acid methyl ester. These features render the S. marcescens ECU1010 lipase attractive for biotechnological applications in the field of organic synthesis and pharmaceutical industry.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, People's Republic China. suxia5664@163.com
authors.forename: Suxia
authors.initials: S
authors.lastname: Li
authors.forename: Kang
authors.initials: K
authors.lastname: Lin
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Pang
authors.forename: Yixin
authors.initials: Y
authors.lastname: Wu
authors.forename: Jianhe
authors.initials: J
authors.lastname: Xu
mesh_terms: D004789:Enzyme Activation; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D002479:Inclusion Bodies / Q000201:enzymology*; D008049:Lipase / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry*; D009930:Organic Chemicals / Q000737:chemistry*; D015202:Protein Engineering / Q000379:methods; D012706:Serratia marcescens / Q000201:enzymology* / Q000235:genetics; D012997:Solvents / Q000737:chemistry*
chemical_list: D009930:Organic Chemicals; D012997:Solvents; D008049:Lipase