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

9 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

Fingerprint

Candida rugosa
Candida
butyrates
Lipase
codons
Codon
hydrolysis
amino acid derivatives
substrate specificity
catalytic activity
esterases
tyrosine
esters
triacylglycerols
assays
Substrates
Protein Isoforms
Hydrolysis
Laurates
Esterases

Keywords

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

ASJC Scopus subject areas

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

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

Characterization of codon-optimized recombinant Candida rugosa Lipase 5 (LIP5). / Lee, Li Chiun; Yen, Chih Chung; Malmis, Conmar C.; Chen, Long Fang; Chen, Jen Chieh; Lee, Guan-Chiun; Shaw, Jei Fu.

In: Journal of Agricultural and Food Chemistry, Vol. 59, No. 19, 12.10.2011, p. 10693-10698.

Research output: Contribution to journalArticle

Lee, Li Chiun ; Yen, Chih Chung ; Malmis, Conmar C. ; Chen, Long Fang ; Chen, Jen Chieh ; Lee, Guan-Chiun ; Shaw, Jei Fu. / Characterization of codon-optimized recombinant Candida rugosa Lipase 5 (LIP5). In: Journal of Agricultural and Food Chemistry. 2011 ; Vol. 59, No. 19. pp. 10693-10698.
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