title: A rapid quantitative activity assay shows that the Vibrio cholerae colonization factor GbpA is an active lytic polysaccharide monooxygenase.
abstract: The discovery of the copper-dependent lytic polysaccharide monooxygenases (LPMOs) has revealed new territory for chemical and biochemical analysis. These unique mononuclear copper enzymes are abundant, suggesting functional diversity beyond their established roles in the depolymerization of biomass polysaccharides. At the same time basic biochemical methods for characterizing LPMOs, such as activity assays are not well developed. Here we describe a method for quantification of C1-oxidized chitooligosaccharides (aldonic acids), and hence LPMO activity. The method was used to quantify the activity of a four-domain LPMO from Vibriocholerae, GbpA, which is a virulence factor with no obvious role in biomass processing.
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 Aas, Norway.
authors.forename: Jennifer S M
authors.initials: JS
authors.lastname: Loose
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 Aas, Norway.
authors.forename: Zarah
authors.initials: Z
authors.lastname: Forsberg
authors.affiliation: Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, Groningen, The Netherlands.
authors.forename: Marco W
authors.initials: MW
authors.lastname: Fraaije
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 Aas, Norway.
authors.forename: Vincent G H
authors.initials: VG
authors.lastname: Eijsink
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, P.O. Box 5003, N-1432 Aas, Norway. Electronic address: gustko@nmbu.no.
authors.forename: Gustav
authors.initials: G
authors.lastname: Vaaje-Kolstad
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002686:Chitin / Q000737:chemistry; D057075:Enzyme Assays / Q000592:standards; D036781:Fimbriae Proteins / Q000737:chemistry; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D010084:Oxidation-Reduction; D012015:Reference Standards; D014734:Vibrio cholerae / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; C022924:colonization factor antigens; D002686:Chitin; D036781:Fimbriae Proteins; D006899:Mixed Function Oxygenases
keywords: AA10; AA11; AA9; Colonization factor; GbpA; Lytic polysaccharide monooxygenase; Vibrio cholerae