Biochemical and catalytic properties of two intracellular β-glucosidases from the fungus Penicillium decumbens active on flavonoid glucosides

In the presence of rutin as sole carbon source, Penicillium decumbens produces two intracellular β-glucosidases named GI and GII, with molecular masses of 56,000 and 460,000 Da, respectively. The two proteins have been purified to homogeneity. GI and GII composed of two and four equal sub-units, respectively and displayed optimal activity at pH 7.0 and temperature 65–75 °C. Both β-glucosidases were competitively inhibited by glucose and glucono-δ-lactone. GI and GII exhibited broad substrate specificity, since they hydrolyzed a range of (1,3)-, (1,4)- and (1,6)-β-glucosides as well as aryl β-glucosides. Determination of kcat/Km revealed that GII hydrolyzed 3–8 times more efficiently the above-mentioned substrates. The ability of GI and GII to deglycosylate various flavonoid glycosides was also investigated. Both enzymes were active against flavonoids glycosylated at the 7 position but GII hydrolyzed them 5 times more efficiently than GI. Of the flavanols tested, both enzymes were incapable of hydrolyzing quercetrin and kaempferol-3-glucoside. The main difference between GI and GII as far as the hydrolysis of flavanols is concerned, was the ability of GII to hydrolyze the quercetin-3-glucoside.