Structure-function characterization reveals new catalytic diversity in the galactose oxidase and glyoxal oxidase family

来自 EBSCO

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作者：

DL Yin，S Urresti，M Lafond，EM Johnston，F Derikvand，L Ciano，JG Berrin，B Henrissat，PH Walton，GJ Davies

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摘要：

Alcohol oxidases, including carbohydrate oxidases, have a long history of research that has generated fundamental biological understanding and biotechnological applications. Despite a long history of study, the galactose 6-oxidase/glyoxal oxidase family of mononuclear copper-radical oxidases, Auxiliary Activity Family 5 (AA5), is currently represented by only very few characterized members. Here we report the recombinant production and detailed structure–function analyses of two homologues from the phytopathogenic fungiColletotrichum graminicolaandC. gloeosporioides,CgrAlcOx andCglAlcOx, respectively, to explore the wider biocatalytic potential in AA5. EPR spectroscopy and crystallographic analysis confirm a common active-site structurevis-à-visthe archetypal galactose 6-oxidase fromFusarium graminearum. Strikingly, however,CgrAlcOx andCglAlcOx are essentially incapable of oxidizing galactose and galactosides, but instead efficiently catalyse the oxidation of diverse aliphatic alcohols. The results highlight the significant potential of prospecting the evolutionary diversity of AA5 to reveal novel enzyme specificities, thereby informing both biology and applications. Auxilliary Activity Family 5 (AA5) comprises mononuclear copper radical oxidases with catalytic diversity that is not well characterised. Here, structural, phylogenetic and biochemical analyses advance our understanding of the potential biological and biotechnology functions of these proteins.

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关键词：

RADICAL-COPPER OXIDASE ARYL-ALCOHOL OXIDASE ACTIVE-SITE PHANEROCHAETE-CHRYSOSPORIUM COLLETOTRICHUM FUNGI PHYLOGENETIC TREES EXPRESSION PURIFICATION OXIDATION SOFTWARE