title: Purification and Characterization of an Extracellular beta-Glucosidase from the Wood-Grown Fungus Xylaria regalis
abstract: Xylaria regalis, a wood-grown ascomycete isolated in Taiwan, produces beta-glucosidase (EC 3.2.1.21) extracellularly. The beta-glucosidase was purified to homogeneity by ammonium sulfate precipitation, ion-exchange, and gel filtration chromatography. The molecular mass of the purified enzyme was estimated to be 85 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With p-nitrophenyl beta-D-glucopyranoside (PNPG) as the substrate at pH 5.0 and 50&deg;C, the Km was 1.72 mM and Vmax was 326 &mgr;mol/min/mg. Optimal activity with PNPG as the substrate was at pH 5.0 and 50&deg;C. The enzyme was stable at pH 5.0 at temperatures up to 50&deg;C. The purified beta-glucosidase was active against PNPG, cellobiose, sophorose, and gentiobiose, but did not hydrolyze lactose, sucrose, Avicel, and o-nitrophenyl beta-D-galactopyranoside. The activity of beta-glucosidase was stimulated by Ca2+, Mg2+, Mn2+, Cd2+ and beta-mercaptoethanol, and inhibited by Ag+, Hg2+, SDS, and p-chloromercuribenzoate (PCMB).
authors.affiliation: Division of Biology, Center for General Education, National Yang-Ming University, Taipei 11221, Taiwan, Republic of China
authors.initials: DL
authors.lastname: Wei
authors.initials: K
authors.lastname: Kirimura
authors.initials: S
authors.lastname: Usami
authors.initials: TH
authors.lastname: Lin