Engineered Bacillus pumilus laccase-like multi-copper oxidase for enhanced oxidation of the lignin model compound guaiacol

Protein Eng Des Sel. 2017 Jun 1;30(6):449-453. doi: 10.1093/protein/gzx026.

Abstract

Laccases and laccase-like multi-copper oxidases (LMCOs) are versatile and robust biocatalysts applied in a variety of oxidative processes, and various studies have attempted to improve their catalytic activity. Here we report the engineering of a bacterial LMCO for enhanced oxidation of the lignin-related compound guaiacol by a combination of structure-guided mutagenesis and DNA shuffling. Mutant L9 showed a 1.39 mM Km for guaiacol and a 2.5-fold increase in turnover rate (kcat/Km = 2.85·104 M-1s-1).

Keywords: DNA shuffling; engineered bacterial LMCO; guaiacol oxidation; laccase engineering; lignin; site-directed mutagenesis.

MeSH terms

  • Bacillus pumilus / enzymology*
  • Bacillus pumilus / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Guaiacol / chemistry
  • Guaiacol / metabolism*
  • Laccase / chemistry
  • Laccase / genetics
  • Laccase / metabolism*
  • Lignin
  • Models, Molecular
  • Oxidation-Reduction
  • Oxidoreductases / chemistry
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism*
  • Protein Engineering
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Guaiacol
  • Lignin
  • Oxidoreductases
  • Laccase
  • copper oxidase