title: A Phanerochaete chrysosporium beta-D-glucosidase/beta-D-xylosidase with specificity for (1-->3)-beta-D-glucan linkages.
abstract: Phanerochaete chrysosporium is the best studied organism with respect to lignin degradation, but its degradation of the xylan component of lignocellulose is only now being studied. When grown on oat spelt xylan (mainly arabinoxylan), it produces an enzyme with beta-D-xylosidase and beta-D-glucosidase activity. This enzyme was purified by ultrafiltration followed by ammonium sulphate precipitation, anion-exchange chromatography using DEAE Biogel and Mono Q, and gel filtration using Superose 12. It is extracellular, with an apparent M(r) value of 44,500 as determined by SDS-PAGE; the pI is 4.67 and activity is maximal at pH 5 and 60 degrees C. The enzyme is of particular interest because its principal activity is against laminaribiose (3-O-beta-D-glucopyranosyl-D-glucopyranose and laminarin [(1-->3)-beta-D-glucan with ca. 3% of beta-(1-->6) branches] rather than cellobiose and xylobiose. It was competitively inhibited by D-glucono-1,5-lactone and deoxynojirimycin; with p-nitrophenyl beta-D-xylopyranoside as substrate, the Ki values were 32 and 87.5 microM, respectively, and with p-nitrophenyl beta-D-glucopyranoside, they were 35 and 68.7 microM, respectively. The Km values with p-nitrophenyl beta-D-xylopyranoside and p-nitrophenyl beta-D-glucopyranoside as substrates were 3.51 and 5.30 mM, respectively.
authors.affiliation: Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, United Kingdom.
authors.forename: J L
authors.initials: JL
authors.lastname: Copa-Patiño
authors.forename: P
authors.initials: P
authors.lastname: Broda
mesh_terms: D000363:Agaricales / Q000201:enzymology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity; D014462:Ultrafiltration; D014995:Xylosidases / Q000302:isolation & purification / Q000378:metabolism*; D001617:beta-Glucosidase / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D005936:Glucans; D014995:Xylosidases; D001617:beta-Glucosidase; C026579:exo-1,4-beta-D-xylosidase