Purification and catalytic properties of novel enantioselective lipase from Acinetobacter sp. ES-1 for hydrolysis of ( S)-ketoprofen ethyl ester

• Published in: Enzyme and microbial technology Vol. 38; no. 3; pp. 443 - 448
• Main Authors: Lee, Kwang-Woo, Bae, Hyun-Ae, Shin, Gab-Sang, Lee, Yong-Hyun
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.02.2006; Elsevier Science
• Subjects: ( R, S)-Ketoprofen ethyl ester; ( S)-Ketoprofen; Acinetobacter; Acinetobacter sp. ES-1; Biological and medical sciences; Biotechnology; Catalytic properties; Enantioselectivity; Fundamental and applied biological sciences. Psychology; Lipase; Acinetobacter sp. ES-1; Catalytic properties; ( R, S)-Ketoprofen ethyl ester; ( S)-Ketoprofen; Enantioselectivity; Lipase; Purification; (S)-Ketoprofen; Enzyme; Neisseriaceae; Triacylglycerol lipase; Enzymatic catalysis; Esterases; Ketoprofen; Carboxylic ester hydrolases; Hydrolysis; (R,S)-Ketoprofen ethyl ester; Ester; Bacteria; Hydrolases; Micrococcales; Acinetobacter
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2005.06.017

A novel enantioselective lipase from Acinetobacter sp. ES-1 exhibiting an excellent enantioselectivity for ( S)-ketoprofen ethyl ester was purified to homogeneity using hydrophobic interaction chromatography. The molecular weight and p I value of the purified enantioselective lipase were 32 kDa and 6.2, respectively, and the optimal temperature and pH were 40 °C and 7.0. The activity was enhanced 1.5-fold in the presence of Ca 2+ ions, and showed the substrate specificity preferably for a medium-chain length, such as triacylglycerides and p-nitrophenyl esters. The N-terminal amino acid residue was A–V–R–S–I–D–G–L–A–I–N–Q–S–A, and no homology was found among other known lipases, indicating a novel type enzyme. The enantioselective hydrolysis of ( R, S)-ketoprofen ethyl ester to optically pure ( S)-ketoprofen was carried out in the aqueous phase reaction system achieving an high enantiomeric excess of 0.99 and conversion yield of 0.49 after 72 h.