Purification and characterization of lipase from Aeromonas sobria LP004.
Lipase from Aeromonas sobria LP004, isolated from raw milk, was purified and characterized. The lipase was purified 10.29 fold to a homogeneous state by ultrafiltration and column chromatography on phenyl sepharose. The molecular weight of the lipase determined by SDS-PAGE was 97 kDa. Purified A. sobria LP004 lipase exhibited the maximum activity at pH 6.0 and 45 degrees C and was stable under alkaline conditions (pH 6.5-10.0) and at temperatures lower than 40 degrees C. This lipase could be classified as a 1,3-position specific enzyme and its catalytic activity was calcium dependent. PMSF, a serine enzyme inhibitor and 2-mercaptoethanol, a reducing agent, did not affect the enzyme activity.
Lotrakul
P
P
Center for Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Dharmsthiti
S
S
D000333:Aeromonas / Q000201:enzymology*; D000818:Animals; D002845:Chromatography; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008892:Milk / Q000382:microbiology; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature; D014105:Tosyl Compounds / Q000494:pharmacology; D014462:Ultrafiltration
D004791:Enzyme Inhibitors; D011480:Protease Inhibitors; D014105:Tosyl Compounds; C003614:4-toluenesulfonyl fluoride; D004492:Edetic Acid; D008049:Lipase