Cloning, Expression, and Biochemical Characterization of an Enantioselective Lipase, YLIP9, from Yarrowia lipolytica MSR80

Appl Biochem Biotechnol. 2015 May;176(1):110-24. doi: 10.1007/s12010-015-1561-y. Epub 2015 Mar 25.

Abstract

A novel lipase gene, ylip9, of Yarrowia lipolytica MSR80 was cloned and expressed in pEZZ18-HB101 system and was 99% identical to YLIP9 of Y. lipolytica CLIB122. It was purified using IgG-Sepharose as ZZ fused YLIP9 and had specific activity of 0.8 U/mg. ZZ-YLIP9 was most active at pH 8.0 and 70 °C. It was stable over a wide pH range of 3.0-11.0 and 100 % active at 70 °C up to 2 h and had t1/2 of 286.42 min at 80 °C. It showed high specificity toward p-nitrophenyldecanoate with kcat and catalytic efficiency of 30.17 s(-1) and 16.67 mM(-1) s(-1), respectively. It was non-regioselective, but an S-enantioselective lipase and the percentage conversion were enhanced in presence of hexane. ZZ-YLIP9 was stable in all of the organic solvents used, and its activity was enhanced by solvents having logP value less than 2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular*
  • Enzyme Stability
  • Fungal Proteins* / biosynthesis
  • Fungal Proteins* / chemistry
  • Fungal Proteins* / genetics
  • Fungal Proteins* / isolation & purification
  • Gene Expression*
  • Lipase* / biosynthesis
  • Lipase* / chemistry
  • Lipase* / genetics
  • Lipase* / isolation & purification
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Yarrowia / enzymology*
  • Yarrowia / genetics

Substances

  • Fungal Proteins
  • Recombinant Fusion Proteins
  • Lipase