Recombinant expression of four oxidoreductases in Phanerochaete chrysosporium improves degradation of phenolic and non-phenolic substrates

J Biotechnol. 2015 Sep 10:209:76-84. doi: 10.1016/j.jbiotec.2015.06.401. Epub 2015 Jun 22.

Abstract

Phanerochaete chrysosporium belongs to a group of lignin-degrading fungi that secretes various oxidoreductive enzymes, including lignin peroxidase (LiP) and manganese peroxidase (MnP). Previously, we demonstrated that the heterologous expression of a versatile peroxidase (VP) in P. chrysosporium recombinant strains is possible. However, the production of laccases (Lac) in this fungus has not been completely demonstrated and remains controversial. In order to investigate if the co-expression of Lac and VP in P. chrysosporium would improve the degradation of phenolic and non-phenolic substrates, we tested the constitutive co-expression of the lacIIIb gene from Trametes versicolor and the vpl2 gene from Pleurotus eryngii, and also the endogenous genes mnp1 and lipH8 by shock wave mediated transformation. The co-overexpression of peroxidases and laccases was improved up to five-fold as compared with wild type species. Transformant strains showed a broad spectrum in phenolic/non-phenolic biotransformation and a high percentage in synthetic dye decolorization in comparison with the parental strain. Our results show that the four enzymes can be constitutively expressed in a single transformant of P. chrysosporium in minimal medium. These data offer new possibilities for an easy and efficient co-expression of laccases and peroxidases in suitable basidiomycete species.

Keywords: Fungal oxidoreductases; Heterologous expression; Laccase; Phanerochaete chrysosporium; Phenolic compounds degradation; Shock waves.

MeSH terms

  • Cloning, Molecular
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Laccase / genetics
  • Laccase / metabolism*
  • Metabolic Engineering / methods
  • Peroxidases / genetics
  • Peroxidases / metabolism*
  • Phanerochaete / enzymology
  • Phanerochaete / genetics*
  • Phanerochaete / metabolism
  • Phenols / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transformation, Genetic

Substances

  • Fungal Proteins
  • Phenols
  • Recombinant Proteins
  • Laccase
  • Peroxidases