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Characterization of a Thermophilic Monosaccharide Stimulated β-Glucosidase from Acidothermus cellulolyticus

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Abstract

The gene(ABK51908) from Acidothermus cellulolyticus encodes a mature protein of 484 residues with a calculated molecular mass of 53.0 kDa. Sequence analysis revealed that the protein had 59% identity to the β-glucosidases CAA82733, which belongs to glycoside hydrolase family 1(GH1). We cloned and expressed the gene in Escherichia coli BL21-Gold(DE3). The recombinant protein(AcBg) had an optimal pH and temperature of 7.0 and 70 °C, respectively. The specific activities of AcBg under optimal conditions were 290 and 33 U/mg for p-nitrophenyl-β-D-glucopyranoside(pNPG) and cellobiose, respectively. AcBg hydrolyzed the oligosaccharide sequentially from the non-reducing end to produce glucose units according to the results of HPLC analysis. AcBg showed high salt tolerance and monosaccharide-stimulation properties. Its activity rose more than 2-fold when 5 mol/L NaCl/KCl were added. The activity of the β-glucosidase was remarkably enhanced in the presence of 0.2 mol/L D-glucose(increased more than 1.9-fold), 0.1 mol/L α-methyl-D-glucose(increased more than 1.4-fold) and 1.0 mol/L D-xylose(increased more than 1.9-fold). The catalysis kinetics and structural changes in various concentrations of glucose were determined. The results indicate that glucose reduces substrate affinity and causes conformational changes.

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Authors and Affiliations

  1. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, School of Life Science, Jilin University, Changchun, 130012, P. R. China

    Yuwei Li, Mingwei Bu, Gui Gao, Yan Feng, Weiwei Han & Zuoming Zhang

  2. State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun, 130012, P. R. China

    Yuwei Li

  3. The Second Hospital of Jilin University, Changchun, 130022, P. R. China

    Peng Chen

  4. College of Food Engineering, Jilin Engineering Normal University, Changchun, 130052, P. R. China

    Xiaohong Li & Changwu Chen

Authors
  1. Yuwei Li
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  2. Mingwei Bu
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  3. Peng Chen
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  4. Xiaohong Li
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  5. Changwu Chen
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  6. Gui Gao
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  7. Yan Feng
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  9. Zuoming Zhang
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Correspondence to Zuoming Zhang.

Additional information

Supported by the National Basic Research Program of China(No.2012CB721003), the Sic-Tech Development Plan of Jilin Province, China(No.20170204040GX) and the Project of the Department of Education of Jilin Province, China(No.2016-438).

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Li, Y., Bu, M., Chen, P. et al. Characterization of a Thermophilic Monosaccharide Stimulated β-Glucosidase from Acidothermus cellulolyticus. Chem. Res. Chin. Univ. 34, 212–220 (2018). https://doi.org/10.1007/s40242-018-7408-7

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  • DOI: https://doi.org/10.1007/s40242-018-7408-7

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