Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of a GH8 β-1,4-Glucanase from Bacillus subtilis B111 and Its Saccharification Potential for Agricultural Straws

  • 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) ;
  • Zhao, Xiaoyan (College of Bioscience and Bioengineering, Jiangxi Agricultural University) ;
  • Wang, Fei (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 : 2021.05.20
  • Accepted : 2021.08.19
  • Published : 2021.10.28
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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℃, respectively, and it was stable at pH 3-9 and temperature ≤50℃. 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

Acknowledgement

This study was supported by funding obtained 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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