Characterization and homologous overexpression of an N-acetylglucosaminidase Nag1 from Trichoderma reesei

Biochem Biophys Res Commun. 2015 Apr 3;459(2):184-188. doi: 10.1016/j.bbrc.2014.12.066. Epub 2014 Dec 19.

Abstract

Trichoderma reesei is thought to be a promising recombinant host for the production and secretion of complex proteins due to its ability to secrete large amounts of proteins. In this study we identified a functional N-acetyl-β-glucosaminidase (NAGase) gene Nag1 in T. reesei. Nag1, a putative gene encoding a GH 20 family NAGase in T. reesei, was cloned and homologous overexpressed in the T. reesei RutC30ΔU3 with a strong cellobiohydrolase1 gene (cbh1) promoter. Nag1 was secreted in its active form and the highest expression level was around 499.85IU/ml. Nag1 has a molecular mass of 80kDa. The optimum pH and temperature were 4.0 and 60°C, respectively.

Keywords: Homologous overexpression; NAGase; Nag1; Trichoderma reesei; cbh1 promoter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosaminidase / chemistry
  • Acetylglucosaminidase / genetics*
  • Acetylglucosaminidase / metabolism*
  • Amino Acid Sequence
  • Cloning, Molecular
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism*
  • Genes, Fungal
  • Hydrogen-Ion Concentration
  • Industrial Microbiology
  • Molecular Sequence Data
  • Molecular Weight
  • Promoter Regions, Genetic
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Temperature
  • Trichoderma / enzymology*
  • Trichoderma / genetics*
  • Up-Regulation

Substances

  • Fungal Proteins
  • Recombinant Proteins
  • Acetylglucosaminidase