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An α-L-fucosidase from Thermus sp. with unusually broad specificity

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Abstract

An α-L-fucosidase (E.C. 3.2.1.51) exhibiting a wide aglycon specificity expressed in ability of cleaving α1 → 6-, α1 →3-, α1 → 4-, and α1 → 2-O-fucosyl bonds in fucosylated oligosaccharides, has been isolated from culture filtrate of Thermus sp. strain Y5. The α-L-fucosidase hydrolyzes p-nitrophenyl α-L-fucopyranoside with V max of 12.0 ± 0.1 μM/min/mg and K m = 0.20 ± 0.05 mM and is able to cleave off about 90% of total L-fucose from pronase-treated fractions of fucosyl-containing glycoproteins and about 30% from the native glycoproteins. The purified enzyme is a tetramer with a molecular mass of 240 ± 10 kDa consisting of four identical subunits with a molecular mass of 61.0 ± 0.5 kDa. The N-terminal sequence showed homology to some α-L-fucosidases from microbial and plant sources. Hydrolysis of p-nitrophenyl α-L-fucopyranoside occurs with retention of the anomeric configuration. Transglycosylating activity of the α-L-fucosidase was demonstrated in reactions with such acceptors as alcohols, N-acetylglucosamine and N-acetylgalactosamine while no transglycosylation products were observed in the reaction with p-nitrophenyl α-L-fucopyranoside. The enzyme can be classified in glycosyl hydrolase family 29.

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Authors and Affiliations

  1. Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute, Russian Academy of Science, 188300, Gatchina, Orlova roscha, Russia

    Elena V. Eneyskaya, Anna A. Kulminskaya, Anna I. Saenko, Olga V. Chepurnaya, Alexander V. Arutyunyan & Konstantin A. Shabalin

  2. Protein Chemistry Laboratory, Institute of Biotechnology, University of Helsinki, Finland

    Nisse Kalkkinen

  3. N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Science, Moscow, Russia

    Nikolay E. Nifantiev & Nilkolay P. Arbatskii

  4. Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute, Russian Academy of Science, 188300, Gatchina, Orlova roscha, Russia

    Kirill N. Neustroev

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  1. Elena V. Eneyskaya
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  4. Nikolay E. Nifantiev
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Eneyskaya, E.V., Kulminskaya, A.A., Kalkkinen, N. et al. An α-L-fucosidase from Thermus sp. with unusually broad specificity. Glycoconj J 18, 827–834 (2001). https://doi.org/10.1023/A:1021163720282

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