1. Characterization of a novel β-L-arabinofuranosidase in Bifidobacterium longum: functional elucidation of a DUF1680 protein family member.
- Author
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Fujita K, Takashi Y, Obuchi E, Kitahara K, and Suganuma T
- Subjects
- Amino Acids metabolism, Arabinose analogs & derivatives, Arabinose chemistry, Arabinose metabolism, Bacterial Proteins isolation & purification, Bifidobacterium growth & development, Chromatography, High Pressure Liquid, Chromatography, Ion Exchange, Chromatography, Thin Layer, Electrophoresis, Polyacrylamide Gel, Fermentation, Glycoproteins metabolism, Glycoside Hydrolases chemistry, Glycosylation, Hydrogen-Ion Concentration, Kinetics, Magnetic Resonance Spectroscopy, Models, Biological, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutant Proteins metabolism, Recombinant Proteins metabolism, Sequence Analysis, Protein, Substrate Specificity, Temperature, Bacterial Proteins chemistry, Bacterial Proteins metabolism, Bifidobacterium enzymology, Glycoside Hydrolases metabolism
- Abstract
Pfam DUF1680 (PF07944) is an uncharacterized protein family conserved in many species of bacteria, actinomycetes, fungi, and plants. Previously, we cloned and characterized the hypBA2 gene as a β-L-arabinobiosidase in Bifidobacterium longum JCM 1217. In this study, we cloned a DUF1680 family member, the hypBA1 gene, which constitutes a gene cluster with hypBA2. HypBA1 is a novel β-L-arabinofuranosidase that liberates L-arabinose from the L-arabinofuranose (Araf)-β1,2-Araf disaccharide. HypBA1 also transglycosylates 1-alkanols with retention of the anomeric configuration. Mutagenesis and azide rescue experiments indicated that Glu-338 is a critical residue for catalytic activity. This study provides the first characterization of a DUF1680 family member, which defines a new family of glycoside hydrolases, the glycoside hydrolase family 127.
- Published
- 2014
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