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

doi:10.1007/s40242-018-7408-7

Chemical Research in Chinese Universities ›› 2018, Vol. 34 ›› Issue (2): 212-220.doi: 10.1007/s40242-018-7408-7

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

LI Yuwei^1,2, BU Mingwei¹, CHEN Peng³, LI Xiaohong⁴, CHEN Changwu⁴, GAO Gui¹, FENG Yan¹, HAN Weiwei¹, ZHANG Zuoming¹

1. Key Laboratory for Molecular Enzymology & Engineering, Ministry of Education, School of Life Science, Jilin University, Changchun 130012, P. R. China;
2. State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, P. R. China;
3. The Second Hospital of Jilin University, Changchun 130022, P. R. China;
4. College of Food Engineering, Jilin Engineering Normal University, Changchun 130052, P. R. China

Received:2017-12-15 Revised:2018-01-25 Online:2018-04-01 Published:2018-02-12
Contact: ZHANG Zuoming,E-mail:zmzhang@jlu.edu.cn E-mail:zmzhang@jlu.edu.cn
Supported by:
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).

Abstract

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℃, 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.

Key words: β-Glucosidase, Monosaccharide-stimulation, Salt tolerance, Thermophilic enzyme

LI Yuwei, BU Mingwei, CHEN Peng, LI Xiaohong, CHEN Changwu, GAO Gui, FENG Yan, HAN Weiwei, ZHANG Zuoming. Characterization of a Thermophilic Monosaccharide Stimulated β-Glucosidase from Acidothermus cellulolyticus[J]. Chemical Research in Chinese Universities, 2018, 34(2): 212-220.

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

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