Systematic Analysis of the Pleurotus ostreatus Laccase Gene (PoLac) Family and Functional Characterization of PoLac2 Involved in the Degradation of Cotton-Straw Lignin

Molecules. 2018 Apr 11;23(4):880. doi: 10.3390/molecules23040880.

Abstract

Fungal laccases play important roles in the degradation of lignocellulose. Although some PoLacs have been reported in several studies, still no comprehensive bioinformatics study of the LAC family in Pleurotus ostreatus has been reported. In this study, we identified 12 laccase genes in the whole genome sequence of P. ostreatus and their physical characteristics, gene distribution, phylogenic relationships, gene structure, conserved motifs, and cis-elements were also analyzed. The expression patterns of 12 PoLac genes at different developmental stages and under different culture substrates were also analyzed. The results revealed that PoLac2 and PoLac12 may be involved in the degradation of lignin and the formation of the fruiting body, respectively. Subsequently, we overexpressed PoLac2 in P. ostreatus by the Agrobacterium tumefaciens-mediated transformation (ATMT) method. The transformants' laccase activity increased in varying degrees, and the gene expression level of PoLac2 in transformants was 2-8 times higher than that of the wild-type strain. Furthermore, the lignin degradation rate by transgenic fungus over 30 days was 2.36-6.3% higher than that of wild-type. Our data show that overexpression of PoLac2 significantly enhanced the lignin degradation of cotton-straw. To our knowledge, this study is the first report to demonstrate the functions of PoLac2 in P. ostreatus.

Keywords: Pleurotus ostreatus; expression profiling; laccase gene; lignin degradation; overexpression; phylogenetic analysis.

MeSH terms

  • Chromosome Mapping
  • Cloning, Molecular / methods*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Fungal
  • Gossypium / chemistry
  • Laccase / genetics*
  • Laccase / metabolism*
  • Lignin / chemistry
  • Multigene Family
  • Phylogeny
  • Pleurotus / enzymology*
  • Pleurotus / genetics
  • Proteolysis

Substances

  • Fungal Proteins
  • Lignin
  • Laccase