Expression and characterization of a cold-active and xylose-stimulated β-glucosidase from Marinomonas MWYL1 in Escherichia coli
The gene encoding a cold-active and xylose-stimulated β-glucosidase of Marinomonas MWYL1 was synthesized and expressed in Escherichia coli. The recombinant enzyme (reBglM1) was purified and characterized. The molecular mass of the purified reBglM1 determined by SDS-PAGE agree with the calculated values (50.6 Da). Optima of temperature and pH for enzyme activity were 40°C and 7.0, respectively. The enzyme exhibited about 20% activity at 5°C and was stable over the range of pH 5.5–10.0. The presence of xylose significantly enhanced enzyme activity even at higher concentrations up to 600 mM, with maximal stimulatory effect (about 1.45-fold) around 300 mM. The enzyme is active with both glucosides and galactosides and showed high catalytic efficiency (kcat = 500.5 s−1) for oNPGlc. These characterizations enable the enzyme to be a promising candidate for industrial applications.
Keywordsβ-Glucosidase Glycoside hydrolase family 1 Marinomonas MWYL1 Cold-active Xylose-stimulated
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