Effective induction, purification and characterization of Trichoderma koningii G-39 beta-xylosidase with high transferase activity

A -xylosidase was induced and purified from the culture filtrate of Trichoderma koningii G-39, grown in a medium containing 1% oat spelts xylan and 0.1% xylose. The presence of xylose unequivocally enhanced the induction of -xylosidase. The purified enzyme, which exhibited a significant -arabinosidase activity, was obtained with high yield simply via ethanol precipitation and a single anion-exchange chromatography and was characterized as a monomeric glycoprotein with an estimated molecular mass of 104 kDa and a pI of 4.6. The Km values towards p-nitrophenyl --xylopyranoside and pnitrophenyl --arabinopyranoside are 0.04 and 7.5 mM, respectively. It is stable at pH 2.5–7.4, 37 °C. The pH and temperature optima are in the range of 3.5–4.0 and 55–60 ° C, respectively. Contrary to most -xylosidases from other sources, Hg2+ (up to 25 mM) has no effect on enzyme activity. Xylose was shown to inhibit the purified enzyme with a moderate Ki value of 5 mM. The enzyme exhibited transxylosylation activity and was characterized as a ʹretainingʹ enzyme, catalysing the hydrolysis of substrate with the retention of anomeric configuration.