title: Characterization of a novel β-glucosidase from a compost microbial metagenome with strong transglycosylation activity.
abstract: The β-glucosidase encoded by the td2f2 gene was isolated from a compost microbial metagenomic library by functional screening. The protein was identified to be a member of the glycoside hydrolase family 1 and was overexpressed in Escherichia coli, purified, and biochemically characterized. The recombinant β-glucosidase, Td2F2, exhibited enzymatic activity with β-glycosidic substrates, with preferences for glucose, fucose, and galactose. Hydrolysis occurred at the nonreducing end and in an exo manner. The order of catalytic efficiency for glucodisaccharides and cellooligosaccharides was sophorose > cellotetraose > cellotriose > laminaribiose > cellobiose > cellopentaose > gentiobiose, respectively. Intriguingly, the p-nitrophenyl-β-D-glucopyranoside hydrolysis activity of Td2F2 was activated by various monosaccharides and sugar alcohols. At a D-glucose concentration of 1000 mM, enzyme activity was 6.7-fold higher than that observed in the absence of D-glucose. With 31.3 mM D-glucose, Td2F2 catalyzed transglycosylation to generate sophorose, laminaribiose, cellobiose, and gentiobiose. Transglycosylation products were detected under all activated conditions, suggesting that the activity enhancement induced by monosaccharides and sugar alcohols may be due to the transglycosylation activity of the enzyme. These results show that Td2F2 obtained from a compost microbial metagenome may be a potent candidate for industrial applications.
authors: National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Japan.
authors: Taku
authors: T
authors: Uchiyama
authors: Kentaro
authors: K
authors: Miyazaki
authors: Katsuro
authors: K
authors: Yaoi
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis / Q000187:drug effects; D002475:Cellobiose / Q000378:metabolism; D004187:Disaccharides / Q000378:metabolism; D005643:Fucose / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D015698:Genomic Library; D005936:Glucans / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006027:Glycosides / Q000378:metabolism; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D054892:Metagenome / Q000235:genetics*; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D012988:Soil Microbiology; D013379:Substrate Specificity; D013402:Sugar Alcohols / Q000494:pharmacology; D013696:Temperature; D001617:beta-Glucosidase / Q000235:genetics* / Q000378:metabolism
chemical_list: D004187:Disaccharides; D005936:Glucans; D006027:Glycosides; D009005:Monosaccharides; D013402:Sugar Alcohols; C014637:laminaribiose; D002475:Cellobiose; D005643:Fucose; D001617:beta-Glucosidase; C100052:gentiobiose; D005947:Glucose; D005690:Galactose; C003328:sophorose
keywords: Biofuel; Enzyme Catalysis; Glycosidases; Glycoside Hydrolases; Metagenomics; Product Inhibition; Transglycosylation; beta-Glucosidase
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