Characterization of a GH8 β-1,4-Glucanase from Bacillus subtilis B111 and Its Saccharification Potential for Agricultural Straws

J Microbiol Biotechnol. 2021 Oct 28;31(10):1446-1454. doi: 10.4014/jmb.2105.05026.

Abstract

Herein, we cloned and expressed an endo-β-1,4-glucanase gene (celA1805) from Bacillus subtilis B111 in Escherichia coli. The recombinant celA1805 contains a glycosyl hydrolase (GH) family 8 domain and shared 76.8% identity with endo-1,4-β-glucanase from Bacillus sp. KSM-330. Results showed that the optimal pH and temperature of celA1805 were 6.0 and 50°C, respectively, and it was stable at pH 3-9 and temperature ≤50°C. Metal ions slightly affected enzyme activity, but chemical agents generally inhibited enzyme activity. Moreover, celA1805 showed a wide substrate specificity to CMC, barley β-glucan, lichenin, chitosan, PASC and avicel. The Km and Vmax values of celA1805 were 1.78 mg/ml and 50.09 μmol/min/mg. When incubated with cellooligosaccharides ranging from cellotriose to cellopentose, celA1805 mainly hydrolyzed cellotetrose (G4) and cellopentose (G5) to cellose (G2) and cellotriose (G3), but hardly hydrolyzed cellotriose. The concentrations of reducing sugars saccharified by celA1805 from wheat straw, rape straw, rice straw, peanut straw, and corn straw were increased by 0.21, 0.51, 0.26, 0.36, and 0.66 mg/ml, respectively. The results obtained in this study suggest potential applications of celA1805 in biomass saccharification.

Keywords: Bacillus subtilis; characterization; endoglucanase; expression; oligosaccharide; saccharification.

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / enzymology*
  • Bacterial Proteins / metabolism
  • Biomass
  • Cellulase / metabolism*
  • Cellulose / analogs & derivatives
  • Cloning, Molecular
  • Enzyme Stability
  • Glucans
  • Hydrolysis
  • Plant Stems
  • Protein Structure, Tertiary
  • Recombinant Proteins
  • Soil Microbiology
  • Substrate Specificity
  • Tetroses

Substances

  • Bacterial Proteins
  • Glucans
  • Recombinant Proteins
  • Tetroses
  • cellotetraose
  • Cellulose
  • Cellulase
  • lichenin