An [alpha]-L-fucosidase from Thermus sp. with unusually broad specificity

• Published in: Glycoconjugate journal 2001-10, Vol.18 (10), p.827-834
• Main Authors: Eneyskaya, Elena V, Kulminskaya, Anna A, Kalkkinen, Nisse, Nifantiev, Nikolay E, Arbatskii, Nilkolay P, Saenko, Anna I, Chepurnaya, Olga V, Arutyunyan, Alexander V, Shabalin, Konstantin A, Neustroev, Kirill N
• Format: Article
• Language: English
• Subjects: Bacteria; Enzymes; Thermus
• ISSN: 0282-0080; 1573-4986
• DOI: 10.1023/A:1021163720282

An α-L-fucosidase (E.C. 3.2.1.51) exhibiting a wide aglycon specificity expressed in ability of cleaving α1 [arrow right] 6-, α1 [arrow right]3-, α1 [arrow right] 4-, and α1 [arrow right] 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^sub max^ of 12.0 ± 0.1 μM/min/mg and K^sub 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.[PUBLICATION ABSTRACT]