title: Characterization of codon-optimized recombinant candida rugosa lipase 5 (LIP5).
abstract: Recombinant Candida rugosa lipase 5 (LIP5) has been functionally expressed along with other isoforms in our laboratory. However, the characterization and codon optimization of LIP5 have not been done. In this work, we characterized, codon-optimized and compared LIP5 with commercial lipase. LIP5 activity on hydrolysis of p-nitrophenyl (p-NP) butyrate was optimal at 55 °C as compared with 37 °C of the commercial lipase. Several assays were also performed to determine the substrate specificity of LIP5. p-NP butyrate (C(4)), butyryl-CoA (C(4)), cholesteryl laurate (C(12)), and N-carbobenzoxy-l-tyrosine-p-nitrophenyl ester (l-NBTNPE) were found as preferred substrates of LIP5. Interestingly, LIP5 specificity on hydrolysis of amino acid-derivative substrates was shown to be the highest among any lipase isoforms, but it had very weak preference on hydrolyzing triacylglycerol substrates. LIP5 also displays a pH-dependent maximum activity of a lipase but an esterase substrate preference in general. The characterization of LIP5 along with that of LIP1-LIP4 previously identified shows that each lipase isoform has a distinct substrate preference and catalytic activity.
authors.affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei, 11529, Taiwan.
authors.forename: Li-Chiun
authors.initials: LC
authors.lastname: Lee
authors.forename: Chih-Chung
authors.initials: CC
authors.lastname: Yen
authors.forename: Conmar C
authors.initials: CC
authors.lastname: Malmis
authors.forename: Long-Fang
authors.initials: LF
authors.lastname: Chen
authors.forename: Jen-Chieh
authors.initials: JC
authors.lastname: Chen
authors.forename: Guan-Chiun
authors.initials: GC
authors.lastname: Lee
authors.forename: Jei-Fu
authors.initials: JF
authors.lastname: Shaw
mesh_terms: D002087:Butyrates / Q000378:metabolism; D002175:Candida / Q000201:enzymology*; D003062:Codon / Q000235:genetics*; D006868:Hydrolysis; D008049:Lipase / Q000235:genetics* / Q000378:metabolism*; D016296:Mutagenesis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002087:Butyrates; D003062:Codon; D011994:Recombinant Proteins; C033592:4-nitrophenyl butyrate; D008049:Lipase