Characterization of Codon-Optimized Recombinant Candida rugosa Lipase 5 (LIP5)

摘要

Recombinant Candida rugosa lipase 5 (LIPS) has been functionally expressed along with other isoforms in our laboratory. However, the characterization and codon optimization of LIPS have not been done. In this work, we characterized, codon-optimized and compared LIPS with commercial lipase. LIP5 activity on hydrolysis of p-mtrophenyl (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 LIPS. p-NP butyrate (C4), butyryl-CoA (C4), cholesteryl laurate (C_(12)), and N-carbobenzoxy-L-ryrosme-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. LLPS 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.