Han, SJ Back, JH Yoon, MY Shin, PK Cheong, CS Sung, MH Hong, SP Chung, IY Han, YS 2024-01-21T09:02:26Z 2024-01-21T09:02:26Z 2021-09-03 2003-05 0300-9084 https://pubs.kist.re.kr/handle/201004/138613 A novel lipase gene, lipase A, of Acinetobacter species SY-01 (A. species SY-01) was cloned, sequenced, and expressed in Bacillus subtilis 168. The deduced amino acid (aa) sequences for the lipase A and its chaperone, lipase-specific chaperone, were found to encode mature proteins of 339 aa (37.2 kDa) and 347 aa (38.1 kDa), respectively. The aa sequence of lipase A and lipase-specific chaperone shared high homology 82 and 67% identity with the lipase A and the lipase B of A. species RAG-1. This new lipase was defined as a group I Proteobacterial lipase family. The expressed lipase A was purified through sequential treatment with Q-Sepharose, Resource Q, and Superdex-S75 columns. The maximal activity was observed at 50 degreesC for hydrolysis of p-nitrophenyl monoesters and found to be stable at pH 9-11, with optimal activity at pH 10. Lipase A hydrolyzed wide range of fatty acid esters of p-nitrophenyl, but preferentially hydrolyzed short length acyl chains (C2 and C4). Moreover, lipase A from A. species SY-01 catalyzed hydrolysis of the two acetate isomers of cis-(+/-)-2-(bromomethyl)-2-(2,4-dichloro phenyl)-1,3-dioxolane-4-methyl acetate, an intermediate required for the synthesis of Itraconazole which was an anti-fungal drug, at different rate and yielded cis-(-)-isomer in 81.5% conversion with 91.9% enantiomeric excess. (C) 2003 Editions scientifiques et medicales Elsevier SAS and Societe francaise de biochimie et biologic moleculaire. All rights reserved. English EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PSEUDOMONAS-GLUMAE LIPASE EXTRACELLULAR LIPASE MOLECULAR-CLONING BACTERIAL LIPASES IN-VITRO SEQUENCE-ANALYSIS ESCHERICHIA-COLI GENE-EXPRESSION PURIFICATION AERUGINOSA Expression and characterization of a novel enantioselective lipase from Acinetobacter species SY-01 Article 10.1016/S0300-9084(03)00057-9 1 BIOCHIMIE, v.85, no.5, pp.501 - 510 BIOCHIMIE 85 5 501 510 scie scopus 000183355900003 2-s2.0-0037809707 Biochemistry & Molecular Biology Biochemistry & Molecular Biology Article PSEUDOMONAS-GLUMAE LIPASE EXTRACELLULAR LIPASE MOLECULAR-CLONING BACTERIAL LIPASES IN-VITRO SEQUENCE-ANALYSIS ESCHERICHIA-COLI GENE-EXPRESSION PURIFICATION AERUGINOSA Acinetobacter species SY-01 lipase lipase-specific chaperone enantioselective cis-(+/-)-2-(bromomethyl)-2-(2,4-dichlorophenyl)-1,3-dioxolane4-methyl acetate