title: Gene cloning, expression, and characterization of the Bacillus amyloliquefaciens PS35 lipase.
abstract: Lipases are enzymes of immense industrial relevance, and, therefore, are being intensely investigated. In an attempt to characterize lipases at molecular level from novel sources, a lipase gene from Bacillus amyloliquefaciens PS35 was cloned, heterologously expressed in Escherichia coli DH5α cells and sequenced. It showed up to 98% homology with other lipase sequences in the NCBI database. The recombinant enzyme was then purified from E. coli culture, resulting in a 19.41-fold purification with 9.7% yield. It displayed a preference for long-chain para-nitrophenyl esters, a characteristic that is typical of true lipases. Its optimum pH and temperature were determined to be 8.0 and 40 °C, respectively. The half-lives were 2.0, 1.0 and 0.5 h at 50 °C, 60 °C and 70 °C, respectively. The metal ions K+ and Fe3+ enhanced the enzyme activity. The enzyme displayed substantial residual activity in the presence of various tested chemical modifiers, and interestingly, the organic solvents, such as n-hexane and toluene, also favored the enzyme activity. Thus, this study involves characterization of B. amyloliquefaciens lipase at molecular level. The key outcomes are novelty of the bacterial source and purification of the enzyme with desirable properties for industrial applications.
authors.affiliation: Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, India.
authors.forename: Palanisamy
authors.initials: P
authors.lastname: Kanmani
authors.affiliation: Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, India.
authors.forename: Kuppamuthu
authors.initials: K
authors.lastname: Kumaresan
authors.affiliation: Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, India.
authors.forename: Jeyaseelan
authors.initials: J
authors.lastname: Aravind
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D008049:Lipase