title: Lipase of Bacillus stratosphericus L1: Cloning, expression and characterization.
abstract: Present work was undertaken to discover new lipolytic enzymes as well as novel bacterial strains for applications in biotechnology. One of the isolated strains identified as Bacillus stratosphericus L1 produced extracellular lipase (LipBST) which was cloned and expressed in Escherichia coli. Purified mature enzyme had a molecular mass of 19kDa. Recombinant protein showed an activity of 6244.5U/mg at pH 9, 35°C. It was stable in the range of 35-55°C and retained more than 60% activity after incubation for 4h. LipBST was activated by organic solvents such as acetone and n-hexane. Lipase was inactivated by all investigated metal ions, inhibitors and detergents. LipBST was determined to be short-chain specific, but also hydrolyzed medium-, long-chain p-nitrophenyl and natural fatty substrates. The values of Vmaxand KMfor p-nitrophenyl butyrate, p-nitrophenyl caprylate, p-nitrophenyl decanoate were 1.1, 2.5, 0.1mMmin-1and 5×10-2, 3.4×10-2, 194×10-2mM, respectively. Biochemical characteristics of LipBST suggest a great potential for various biotechnological applications including detergent formulation, bioremediation and organic synthesis processes.
authors.affiliation: Department of Microbiology and Biotechnology, Faculty of Natural Science, Vilnius University, 21/27, M. K. Čiurlionio Street, LT-03101, Vilnius, Lithuania. Electronic address: alisa.gricajeva@gmail.com.
authors.forename: Alisa
authors.initials: A
authors.lastname: Gricajeva
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Natural Science, Vilnius University, 21M.K. Čiurlionio Street, LT-03101, Vilnius, Lithuania.
authors.forename: Vida
authors.initials: V
authors.lastname: Bendikienė
authors.affiliation: Department of Microbiology and Biotechnology, Faculty of Natural Science, Vilnius University, 21/27, M. K. Čiurlionio Street, LT-03101, Vilnius, Lithuania.
authors.forename: Lilija
authors.initials: L
authors.lastname: Kalėdienė
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D015870:Gene Expression; D008049:Lipase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D008049:Lipase
keywords: Bacillus stratosphericus; Expression; Kinetics; Lipase; Purification