Cloning of a novel lipase gene, lipJ08, from Candida rugosa and expression in Pichia pastoris by codon optimization

Biotechnol Lett. 2010 Feb;32(2):269-76. doi: 10.1007/s10529-009-0141-z. Epub 2009 Oct 17.

Abstract

A novel lipase gene, lipJ08, was cloned from Candida rugosa ATCC14830, along with the already reported five lipase genes (lip1-lip5). Nucleotide sequencing indicated that the lipJ08 gene contains a 1650 bp open reading frame (ORF) without introns. The deduced amino acid sequence corresponds to 534 amino acid residues, including a putative signal sequence of 15 amino acid residues. Seventeen of the non-universal serine codons (CTG) of lipJ08 were converted into universal serine codons (TCT) by PCR-based mutagenesis. The native and codon-optimized lipJ08 genes were expressed in Pichia pastoris. The hydrolytic activity of the recombinant LIPJ08 was 4.7 U/ml, whereas the activity of the recombinant wild-type lipase could not be detected.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Candida / enzymology*
  • Candida / genetics*
  • Cloning, Molecular / methods*
  • Codon
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism*
  • Lipase / genetics*
  • Lipase / metabolism*
  • Pichia / enzymology*
  • Pichia / genetics*
  • Protein Engineering / methods*
  • Recombinant Proteins / metabolism

Substances

  • Codon
  • Fungal Proteins
  • Recombinant Proteins
  • Lip4 protein, Candida rugosa
  • Lipase