title: Comparative NMR analysis of cellooligosaccharide hydrolysis by GH9 bacterial and plant endo-1,4-beta-glucanases.
abstract: 1H NMR spectroscopy has been used to analyze the product profiles arising from the hydrolysis of cellooligosaccharides by family GH9 cellulases. The product profiles obtained with the wild type and several active site mutants of a bacterial processive endoglucanase, TfCel9A, were compared with those obtained by a randomly acting plant endoglucanase, PttCel9A. PttCel9A is an orthologue of the Arabidopsis endocellulase, Korrigan, which is required for efficient cellulose biosynthesis. As expected, poplar PttCel9A was shown to catalyze the degradation of cellooligosaccharides by inversion of the configuration of the anomeric carbon. The product analyses showed that the number of interactions between the glucose units of the substrate and the aromatic residues in the enzyme active sites determines the point of cleavage in both enzymes.
authors.affiliation: KTH Biotechnology, Swedish Center for Biomimetic Fiber Engineering, AlbaNova, SE-10691 Stockholm, Sweden.
authors.forename: Ulla J
authors.initials: UJ
authors.lastname: Rudsander
authors.forename: Corine
authors.initials: C
authors.lastname: Sandstrom
authors.forename: Kathleen
authors.initials: K
authors.lastname: Piens
authors.forename: Emma R
authors.initials: ER
authors.lastname: Master
authors.forename: David B
authors.initials: DB
authors.lastname: Wilson
authors.forename: Harry
authors.initials: H
authors.lastname: Brumer Iii
authors.forename: Lennart
authors.initials: L
authors.lastname: Kenne
authors.forename: Tuula T
authors.initials: TT
authors.lastname: Teeri
mesh_terms: D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics; D002480:Cellulase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008954:Models, Biological; D015394:Molecular Structure; D009154:Mutation / Q000235:genetics; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism*; D032107:Populus / Q000201:enzymology*
chemical_list: D009844:Oligosaccharides; D002480:Cellulase