title: Kinetic properties and contribution to cellulose saccharification of a cloned Pseudomonas beta-glucosidase.
abstract: The plasmid pND71, which encodes beta-glucosidase (cellobiase) activity, cloned from the cellulolytic Pseudomonad, PS2-2, was mobilized by conjugation into 10 Pseudomonas strains. The highest specific activity was produced by 17498 (pND71) and the properties of the enzyme produced from this transconjugant were studied. The enzyme was shown to be cell associated, to have a temperature optimum of 37 degrees C, a pH optimum of 7.0 and Km values of 1.33 and 2.94 mM for pNPG and cellobiose respectively. It was competitively inhibited by glucose, with a Ki of 30 mM. Evidence was obtained which suggested that the enzyme was produced constitutively and that synthesis was not repressed by glucose. When culture preparations were used in combination with Trichoderma reesei QM9414 and C30 enzyme preparations to saccharify cellulose, 17498 (pND71) was more effective than preparations of PS2-2 in acting synergistically with T. reesei to solubilize more carbohydrate and produce more glucose.
authors.affiliation: Department of Biotechnology, University of New South Wales.
authors.forename: P A
authors.initials: PA
authors.lastname: Rickard
authors.forename: B A
authors.initials: BA
authors.lastname: Ghani
authors.forename: R J
authors.initials: RJ
authors.lastname: Lucas
authors.forename: N W
authors.initials: NW
authors.lastname: Dunn
mesh_terms: D050260:Carbohydrate Metabolism; D002482:Cellulose / Q000378:metabolism*; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004795:Enzyme Stability; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D010957:Plasmids; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
chemical_list: D002482:Cellulose; D001617:beta-Glucosidase; D005947:Glucose