Characterization of Codon-Optimized Recombinant Candida rugosa Lipase 5 (LIP5)
Resource
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 59(19), 10693-10698
Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Journal Volume
59
Journal Issue
19
Pages
10693-10698
Date Issued
2011
Date
2011
Author(s)
Lee, Li-Chiun
Yen, Chih-Chung
Malmis, Conmar C.
Chen, Long-Fang
Chen, Jen-Chieh
Lee, Guan-Chiun
Shaw, Jei-Fu
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.
Type
journal article
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