title: Expression and biochemical characterization of a thermophilic organic solvent-tolerant lipase from Bacillus sp. DR90.
abstract: The objective of the present study was the isolation, molecular cloning and biochemical characterization of a thermophilic organic solvent-resistant lipase from Bacillus sp. DR90. The lipase gene was expressed in Escherichia coli BL21(DE3) using pET-28a(+) vector. The purification of recombinant lipase was conducted by nickel affinity chromatography and its biochemical properties were determined. The lipase sequence with an ORF of 639 bp contains the conserved pentapeptide Ala-His-Ser-Met-Gly. His-tagged recombinant lipase had a specific activity of 1,126 U/mg with a molecular mass of 26.8 kDa. The cloned lipase was optimally active at pH 8.0 and 75 °C representing high stability in broad ranges of temperature and pH. High performance liquid chromatography was used to determine the major compounds released during the lipase-catalyzed reaction of p-nitrophenyl derivatives as well as the substrate specificity. The purified lipase showed high compatibility towards various organic solvents, surfactants and commercial solid/liquid detergents; therefore the recombinant DR90 lipase could be considered as a probable candidate for future applications, predominantly in detergent processing industries.
authors: Department of Chemistry, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran, asoodeh@um.ac.ir.
authors: Ahmad
authors: A
authors: Asoodeh
authors: Shirin
authors: S
authors: Emtenani
authors: Shamsi
authors: S
authors: Emtenani
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019216:Metals, Heavy / Q000494:pharmacology; D008969:Molecular Sequence Data; D009930:Organic Chemicals; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
chemical_list: D019216:Metals, Heavy; D009930:Organic Chemicals; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D008049:Lipase