An alpha-L-fucosidase from Thermus sp. with unusually broad specificity.
An alpha-L-fucosidase (E.C. 3.2.1.51) exhibiting a wide aglycon specificity expressed in ability of cleaving alpha1 --> 6-, alpha1 -->3-, alpha1 --> 4-, and alpha1 --> 2-O-fucosyl bonds in fucosylated oligosaccharides, has been isolated from culture filtrate of Thermus sp. strain Y5. The alpha-L-fucosidase hydrolyzes p-nitrophenyl alpha-L-fucopyranoside with V(max) of 12.0 +/- 0.1 microM/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 alpha-L-fucosidases from microbial and plant sources. Hydrolysis of p-nitrophenyl alpha-L-fucopyranoside occurs with retention of the anomeric configuration. Transglycosylating activity of the alpha-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 alpha-L-fucopyranoside. The enzyme can be classified in glycosyl hydrolase family 29.
Eneyskaya
E V
EV
Molecular and Radiation Biophysics Division, Petersburg Nuclear Physics Institute, Russian Academy of Science, 188300 Gatchina, Orlova roscha, Russia.
Kulminskaya
A A
AA
Kalkkinen
N
N
Nifantiev
N E
NE
Arbatskii
N P
NP
Saenko
A I
AI
Chepurnaya
O V
OV
Arutyunyan
A V
AV
Shabalin
K A
KA
Neustroev
K N
KN
D000595:Amino Acid Sequence; D002845:Chromatography / Q000379:methods; D017124:Conserved Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005643:Fucose / Q000096:biosynthesis / Q000737:chemistry; D006031:Glycosylation; D007700:Kinetics; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D013379:Substrate Specificity; D013824:Thermus / Q000201:enzymology*; D014462:Ultrafiltration; D005644:alpha-L-Fucosidase / Q000302:isolation & purification / Q000378:metabolism*
D009844:Oligosaccharides; D005643:Fucose; D005644:alpha-L-Fucosidase