Cloning, purification and study of recombinant GH3 family β-glucosidase from Penicillium verruculosum

Biochimie. 2020 Jan:168:231-240. doi: 10.1016/j.biochi.2019.11.009. Epub 2019 Nov 19.

Abstract

A novel bgl1 gene, encoding GH3 family β-glucosidase from Penicillium verruculosum (PvBGL), was cloned and heterologously expressed in P. canescens RN3-11-7 (niaD-) strain under the control of the strong xylA gene promoter. The recombinant rPvBGL was purified and their properties were studied in comparison with those of rAnBGL from Aspergillus niger expressed previously in the same fungal host. The rPvBGL had an observed molecular mass of 90 kDa (SDS-PAGE data) and displayed the enzyme maximum activity at pH 4.6 and 65 °C. The enzyme half-life time at 60 °C was found to be 87 min. Unlike the rAnBGL, the rPvBGL was not adsorbed on microcrystalline cellulose, which gives the latter enzyme an advantage in cellulose conversion with a longer time of hydrolysis.

Keywords: Aspergillus niger; Cellobiose; Hydrolysis; Inhibition; Penicillium verruculosum; β-Glucosidase.

Publication types

  • Comparative Study

MeSH terms

  • Aspergillus niger / enzymology*
  • Cellulose / chemistry
  • Cloning, Molecular
  • Fungal Proteins* / chemistry
  • Fungal Proteins* / isolation & purification
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Kinetics
  • Molecular Weight
  • Penicillium / enzymology*
  • Recombinant Proteins* / chemistry
  • Recombinant Proteins* / isolation & purification
  • Substrate Specificity
  • beta-Glucosidase* / chemistry
  • beta-Glucosidase* / isolation & purification

Substances

  • Fungal Proteins
  • Recombinant Proteins
  • Cellulose
  • beta-Glucosidase
  • microcrystalline cellulose