Secretory expression, characterization and docking study of glucose-tolerant β-glucosidase from B. subtilis

International Journal of Biological Macromolecules
Shivangi ChamoliAshok Kumar Verma

Abstract

The thermostable, glucose tolerant β-glucosidase gene (bgl) of Glycoside hydrolase family 1, isolated from Bacillus subtilis, was cloned and overexpressed in Escherichia coli. The bgl has open reading frame of 1,407 bp, encoding 469 amino acids with predicted molecular weight of 53 kDa. The recombinant protein (BGL) was purified 10.76 fold to homogeneity with specific activity of 54.04U/mg and recovery of 38.67%. The purified BGL was optimally active at pH 6.0 and temperature 60°C. The enzyme retained more than 85% of maximum activity after 1h preincubation at 60°C. The kinetic analysis indicated that BGL has highest catalytic efficiency (Kcat/Km) against p-nitrophenyl-β-d-xylopyranoside (654.58 mM(-1)s(-1)) followed by p-nitrophenyl-β-d-glucopyranoside (292.53 mM(-1)s(-1)) and p-nitrophenyl-β-d-galactopyranoside (61.17 mM(-1)s(-1)). The Ki value for glucose and δ-gluconolactone was determined to be 1.9 mM and 0.018 mM, respectively. The BGL exhibited high tolerance against detergents and organic solvents. The homology modeling revealed that protein has 19 α-helices and 4 β-sheets and adopted (α/β)8 TIM barrel structure. Substrate docking and LigPlot analysis depicted the amino acids of active site involved in hydrogen bonding ...Continue Reading

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Citations

Feb 29, 2020·Journal of Biomolecular Structure & Dynamics·Leonardo Henrique Franca de LimaKlaus Roman Liedl
Jan 18, 2019·International Journal of Molecular Sciences·Diego César Batista MarianoRaquel Cardoso de Melo-Minardi
Jul 6, 2020·Applied Biochemistry and Biotechnology·Jingjing SunJianhua Hao
Sep 7, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Leon Sulfierry Corrêa CostaLeonardo Henrique Franca de Lima
Jul 3, 2020·BMC Molecular and Cell Biology·Diego MarianoRaquel Cardoso de Melo-Minardi

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