Molecular cloning, expression, purification and characterization of a novel cellulase gene (Bh-EGaseI) in the beetle Batocera horsfieldi

Gene. 2016 Jan 15;576(1 Pt 1):45-51. doi: 10.1016/j.gene.2015.09.057. Epub 2015 Sep 26.

Abstract

Wood-feeding insects depend heavily on the secretion of a combination of cellulases, mainly endoglucanases and other glucanases such as exoglucanases and xylanases, to achieve efficient digestion of the cellulose of cellulosic materials. In this paper, we report a novel cellulose Bh-EGaseI belonging to the glycoside hydrolase family 45(gh45-1) obtained from the beetle Batocera horsfieldi. The Bh-EGaseI gene spans 714 bp and consists of three exons coding 237 amino acid residues. The cDNA encoding Bh-EGaseI was expressed as 25 KDa in baculovirus-infected Bombyx mori larvae. The expression products of Bh-EGaseI from larval hemolymph showed a specific enzymatic activity of approximately 1030.87 IU per mg. The enzyme was active over a wide range of pH and temperatures; optimal activity was observed at 40 °C and pH 4.0. The effects of ions on Bh-EGaseI activity were also studied, and results indicated that activity decreased to different extents upon addition of ions. Investigations on Bh-EGaseI facilitate their potential application in the production of bioenergy and biomaterials from cellulosic biomass in the future.

Keywords: Baculovirus; Batocera horsfieldi; Cellulase; Endoglucanases; Silkworm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cellulase* / biosynthesis
  • Cellulase* / chemistry
  • Cellulase* / genetics
  • Cellulase* / isolation & purification
  • Cloning, Molecular
  • Coleoptera* / enzymology
  • Coleoptera* / genetics
  • Gene Expression
  • Insect Proteins* / biosynthesis
  • Insect Proteins* / chemistry
  • Insect Proteins* / genetics
  • Insect Proteins* / isolation & purification
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification

Substances

  • Insect Proteins
  • Recombinant Proteins
  • Cellulase