Characteristics and kinetic analysis of β-glucosidase (MaBgl) from Mucor ardhlaengiktus RSC1

Abstract

Cellulolytic enzymes from fungi are complex compound enzyme structures that generally include three kinds of hydrolytic enzymes, which are called endo-β-1,4-glucanases, exo-β-1,4-glucanases, and β-1,4-glucosidases (β-glucosidases). The fungus Mucor ardhlaengiktus isolated from field rice straw produced a β-glucosidase (MaBgl). Maximal MaBgl production occurred when M. ardhlaengiktus was incubated for 6 days in fermentation liquor at 30 °C and an initial pH of 6. Purified MaBgl was obtained from M. ardhlaengiktus fermentation broth by ammonium sulfate fractional precipitation and DEAE-Sepharose FF ion exchange chromatography. The molecular weight of MaBgl as determined by SDS-PAGE electrophoresis was approximately 72 kDa. The kinetic parameters, Michaelis constant (Km) and maximum velocity (Vmax), of MaBgl were 78.2 μmol/L and 28.5 μmol/(L·min), respectively. Assays of MaBgl produced by M. ardhlaengiktus RSC1 under different conditions were investigated by the 3,5-dinitrosalicylic acid (DNS) assay for glucose. The pH and temperature optima for catalytic activity of MaBgl were pH 4.8 and 50 °C, respectively. MaBgl exhibited good thermal stability in the range of 20 to 30 °C, but the thermal stability of MaBgl decreased rapidly over 60 °C. MaBgl had better pH stability between pH 4.6 and 5.0, and the stability of MaBgl decreased when the pH value was lower or higher than this range.