title: Gene cloning and catalytic characterization of cold-adapted lipase of Photobacterium sp. MA1-3 isolated from blood clam.
abstract: A lipase-producing Photobacterium strain (MA1-3) was isolated from the intestine of a blood clam caught at Namhae, Korea. The lipase gene was cloned by shotgun cloning and encoded 340 amino acids with a molecular mass of 38,015 Da. It had a very low sequence identity with other bacterial lipases, with the exception of that of Photobacterium lipolyticum M37 (83.2%). The MA1-3 lipase was produced in soluble form when Escherichia coli cells harboring the gene were cultured at 18°C. Its optimum temperature and pH were 45°C and pH 8.5, respectively. Its activation energy was calculated to be 2.69 kcal/mol, suggesting it to be a cold-adapted lipase. Its optimum temperature, temperature stability, and substrate specificity were quite different from those of M37 lipase, despite the considerable sequence similarities. Meanwhile, MA1-3 lipase performed a transesterification reaction using olive oil and various alcohols including methanol, ethanol, 1-propanol, and 1-butanol. In the presence of t-butanol as a co-solvent, this lipase produced biodiesel using methanol and plant or waste oils. The highest biodiesel conversion yield (73%) was achieved using waste soybean oil and methanol at a molar ratio of 1:5 after 12 h using 5 units of lipase.
authors: Biotechnology Research Division, National Fisheries Research and Development Institute, 408-1 Gijang, Busan 619-902, Republic of Korea.
authors: Young Ok
authors: YO
authors: Kim
authors: Vivia
authors: V
authors: Khosasih
authors: Bo-Hye
authors: BH
authors: Nam
authors: Sang-Jun
authors: SJ
authors: Lee
authors: Antonius
authors: A
authors: Suwanto
authors: Hyung Kwoun
authors: HK
authors: Kim
mesh_terms: D000438:Alcohols / Q000378:metabolism; D000818:Animals; D055162:Biocatalysis; D056804:Biofuels / Q000600:supply & distribution*; D049872:Bivalvia / Q000382:microbiology*; D003001:Cloning, Molecular; D003080:Cold Temperature; D004795:Enzyme Stability; D004951:Esterification; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000432:Methanol / Q000378:metabolism; D008970:Molecular Weight; D000069463:Olive Oil; D010776:Photobacterium / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D010938:Plant Oils / Q000378:metabolism; D012997:Solvents; D013024:Soybean Oil / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000438:Alcohols; D056804:Biofuels; D000069463:Olive Oil; D010938:Plant Oils; D012997:Solvents; D013024:Soybean Oil; D008049:Lipase; D000432:Methanol