期刊
BIOCHIMIE
卷 132, 期 -, 页码 54-65出版社
ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
DOI: 10.1016/j.biochi.2016.10.014
关键词
alpha-L-fucosidase; Fusarium proliferatum; alpha-(1 -> 4)-L-fucosidic linkage; alpha-L-fucobiosides; Translycosylation
资金
- Russian Foundation for Basic Research (RFBR) [16-38-00881]
- Russian Science Foundation (RSF) [16-14-00109, 14-50-00126, 14-25-00132]
- state grant [17.1360.2014/K]
- RFBR [16-38-00881]
- RSF [16-14-00109, 14-50-00126, 14-25-00132]
- Russian Science Foundation [16-14-00109] Funding Source: Russian Science Foundation
Here, we report the biochemical characterization of a novel alpha-L-fucosidase with broad substrate specificity (FpFucA) isolated from the mycelial fungus Fusarium proliferatum LE1. Highly purified alpha-L-fucosidase was obtained from several chromatographic steps after growth in the presence of L-fucose. The purified alpha-L-fucosidase appeared to be a monomeric protein of 67 +/- 1 kDa that was able to hydrolyze the synthetic substrate p-nitrophenyl alpha-L-fucopyranoside (pNPFuc), with K-m = 1.1 +/- 0.1 mM and k(cat) = 39.8 +/- 1.8 s(-1). L-fucose, 1-deoxyfuconojirimycin and tris(hydroxymethyl)aminomethane inhibited pNPFuc hydrolysis, with inhibition constants of 0.2 +/- 0.05 mM, 7.1 +/- 0.05 nM, and 12.2 +/- 0.1 mM, respectively. We assumed that the enzyme belongs to subfamily A of the GH29 family (CAZy database) based on its ability to hydrolyze practically all fucose-containing oligosaccharides used in the study and the phylogenetic analysis. We found that this enzyme was a unique alpha-L-fucosidase that preferentially hydrolyzes the alpha-(1 -> 4)-L-fucosidic linkage present in alpha-L-fucobiosides with different types of linkages. As a retaining glycosidase, FpFucA is capable of catalyzing the transglycosylation reaction with alcohols (methanol, ethanol, and 1-propanol) and pNP-containing monosaccharides as acceptors. These features make the enzyme an important tool that can be used in the various modifications of valuable fucose-containing compounds. (C) 2016 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.
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