Engineering the expression and biochemical characteristics of recombinant candida rugosa LIP2 lipase by removing the additional n-terminal peptide and regional codon optimization

Shu Wei Chang, Chia Fang Li, Guan-Chiun Lee, Tzunuan Yeh, Jei Fu Shaw

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Candida rugosa lipase (CRL), an important industrial enzyme, has been established, containing several different isoforms which were encoded by the high-identity lip gene family (lip1 to lip7). In this study, we compared the expression and biochemical characterization with three different engineered lip2 constructions in the yeast Pichia pastoris. Our results showed that lip2 (lip2) has an overall improvement of 50% higher production yield (1.446 U/mL) relative to that of nflip2 (0.964 U/mL) at 7 days of cultivation time. Codon-optimized lip2 (colip2) has a 2.3-fold higher production yield (2.182 U/mL) compared to that of lip2 (noncodon-optimized; 1.446 U/mL) and nflip2 (0.964 U/mL), with a cultivation time of 5 days. This finding demonstrated that the removal of the N-terminus and the regional codon optimization of the lip2 gene fragment at the 5′ end can greatly increase the expression level of recombinant LIP2 in the P. pastoris system. The distinct biochemical properties of our purified recombinant nfLIP2 and LIP2 suggested that they are potentially useful for various industrial applications.

Original languageEnglish
Pages (from-to)6710-6719
Number of pages10
JournalJournal of Agricultural and Food Chemistry
Volume59
Issue number12
DOIs
Publication statusPublished - 2011 Jun 22

Fingerprint

Candida rugosa
Pichia pastoris
Candida
Lipase
codons
Codon
engineering
Genes
peptides
Peptides
Pichia
industrial applications
lips
Lip
Yeast
Industrial applications
Protein Isoforms
genes
Yeasts
yeasts

Keywords

  • Candida rugosa lipase
  • N-terminal peptide
  • Pichia pastoris
  • codon optimization
  • isoforms

ASJC Scopus subject areas

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

Cite this

Engineering the expression and biochemical characteristics of recombinant candida rugosa LIP2 lipase by removing the additional n-terminal peptide and regional codon optimization. / Chang, Shu Wei; Li, Chia Fang; Lee, Guan-Chiun; Yeh, Tzunuan; Shaw, Jei Fu.

In: Journal of Agricultural and Food Chemistry, Vol. 59, No. 12, 22.06.2011, p. 6710-6719.

Research output: Contribution to journalArticle

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