Chemical Research in Chinese Universities

, Volume 34, Issue 2, pp 212–220 | Cite as

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

  • Yuwei Li
  • Mingwei Bu
  • Peng Chen
  • Xiaohong Li
  • Changwu Chen
  • Gui Gao
  • Yan Feng
  • Weiwei Han
  • Zuoming Zhang
Article
  • 6 Downloads

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.

Keywords

β-Glucosidase Monosaccharide-stimulation Salt tolerance Thermophilic enzyme 

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Copyright information

© Jilin University, The Editorial Department of Chemical Research in Chinese Universities and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yuwei Li
    • 1
    • 2
  • Mingwei Bu
    • 1
  • Peng Chen
    • 3
  • Xiaohong Li
    • 4
  • Changwu Chen
    • 4
  • Gui Gao
    • 1
  • Yan Feng
    • 1
  • Weiwei Han
    • 1
  • Zuoming Zhang
    • 1
  1. 1.Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, School of Life ScienceJilin UniversityChangchunP. R. China
  2. 2.State Key Laboratory of Supramolecular Structure and Materials, College of ChemistryJilin UniversityChangchunP. R. China
  3. 3.The Second Hospital of Jilin UniversityChangchunP. R. China
  4. 4.College of Food EngineeringJilin Engineering Normal UniversityChangchunP. R. China

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