Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

  • Huang, Zhen (Key Laboratory of Animal Nutrition of Jiangxi Province, Nutritional Feed Development Engineering Research Center, Jiangxi Agricultural University) ;
  • Ni, Guorong (College of Land Resources and Environment, Jiangxi Agricultural University) ;
  • Wang, Fei (College of Bioscience and Bioengineering, Jiangxi Agricultural University) ;
  • Zhao, Xiaoyan (College of Bioscience and Bioengineering, Jiangxi Agricultural University) ;
  • Chen, Yunda (College of Bioscience and Bioengineering, Jiangxi Agricultural University) ;
  • Zhang, Lixia (College of Bioscience and Bioengineering, Jiangxi Agricultural University) ;
  • Qu, Mingren (Key Laboratory of Animal Nutrition of Jiangxi Province, Nutritional Feed Development Engineering Research Center, Jiangxi Agricultural University)
  • Received : 2021.11.10
  • Accepted : 2021.12.13
  • Published : 2022.04.28
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Abstract

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℃ 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

Acknowledgement

This research was supported by funding from China Agriculture Research System of MOF and MARA (CARS-37) and the Postgraduate Innovation Special Foundation of Jiangxi Province (YC2020-B087).

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