Characterization of codon-optimized recombinant Candida rugosa Lipase 5 (LIP5)

Li Chiun Lee, Chih Chung Yen, Conmar C. Malmis, Long Fang Chen, Jen Chieh Chen, Guan-Chiun Lee, Jei Fu Shaw

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)10693-10698
Number of pages6
JournalJournal of Agricultural and Food Chemistry
Volume59
Issue number19
DOIs
Publication statusPublished - 2011 Oct 12

Keywords

  • Candida rugosa lipase 5 (LIP5)
  • hydrolysis
  • specificity
  • substrate

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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  • Cite this

    Lee, L. C., Yen, C. C., Malmis, C. C., Chen, L. F., Chen, J. C., Lee, G-C., & Shaw, J. F. (2011). Characterization of codon-optimized recombinant Candida rugosa Lipase 5 (LIP5). Journal of Agricultural and Food Chemistry, 59(19), 10693-10698. https://doi.org/10.1021/jf202161a