Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

J Microbiol Biotechnol. 2022 Apr 28;32(4):484-492. doi: 10.4014/jmb.2111.11017.


Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70°C for 1 h. The Km and Vmax of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.

Keywords: Bacillus subtilis; characterization; expression; lichenase; oligosaccharide; β-glucan.

MeSH terms

  • Bacillus subtilis* / genetics
  • Bacillus subtilis* / metabolism
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Glycoside Hydrolases / metabolism
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Oligosaccharides / metabolism
  • Substrate Specificity
  • beta-Glucans* / metabolism


  • Oligosaccharides
  • beta-Glucans
  • Glycoside Hydrolases
  • licheninase