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Literature summary for 3.2.1.63 extracted from

  • Nagae, M.; Tsuchiya, A.; Katayama, T.; Yamamoto, K.; Wakatsuki, S.; Kato, R.
    Structural basis of the catalytic reaction mechanism of novel 1,2-alpha-L-fucosidase from Bifidobacterium bifidum (2007), J. Biol. Chem., 282, 18497-18509.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
Sodium azide for D766A mutant only Bifidobacterium bifidum

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli, selenomethionine-substituted protein Bifidobacterium bifidum

Crystallization (Commentary)

Crystallization (Comment) Organism
of the AfcA catalytic domain in unliganded and complexed forms with the inhibitor deoxyfuconojirimycin, the substrate 2’-fucosyllactose, and the products L-fucose and lactose at 1.12-2.10 A resolution Bifidobacterium bifidum

Protein Variants

Protein Variants Comment Organism
D766A site directed mutagenesis Bifidobacterium bifidum
D766E site directed mutagenesis Bifidobacterium bifidum
E485A site directed mutagenesis Bifidobacterium bifidum
E566A site directed mutagenesis Bifidobacterium bifidum
N421A site directed mutagenesis Bifidobacterium bifidum
N421G site directed mutagenesis Bifidobacterium bifidum
N421G/N423G site directed mutagenesis Bifidobacterium bifidum
N423D site directed mutagenesis Bifidobacterium bifidum
N423G site directed mutagenesis Bifidobacterium bifidum

Inhibitors

Inhibitors Comment Organism Structure
deoxyfuconojirimycin
-
Bifidobacterium bifidum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.035
-
2-fucosyllactose N421A mutant Bifidobacterium bifidum
0.056
-
2-fucosyllactose N421G mutant Bifidobacterium bifidum
0.075
-
2-fucosyllactose N423G mutant Bifidobacterium bifidum
0.089
-
2-fucosyllactose D766A mutant Bifidobacterium bifidum
0.091
-
2-fucosyllactose wild type Bifidobacterium bifidum
0.16
-
2-fucosyllactose E566A mutant Bifidobacterium bifidum
0.19
-
2-fucosyllactose D766E mutant Bifidobacterium bifidum
0.32
-
2-fucosyllactose N423D mutant Bifidobacterium bifidum
17
-
2-fucosyllactose E485A mutant Bifidobacterium bifidum

Organism

Organism UniProt Comment Textmining
Bifidobacterium bifidum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
of the recombinant protein by saturated ammonium sulfate precipitation and Q sepharose fast flow, CHT ceramic hydroxyapatite and gel filtration column chromatography Bifidobacterium bifidum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2'-fucosyllactose + H2O
-
Bifidobacterium bifidum fucose + lactose
-
?
methyl-2-alpha-L-fucopyranosyl-beta-D-galactoside + H2O
-
Bifidobacterium bifidum L-fucose + methyl beta-D-galactoside
-
?

Synonyms

Synonyms Comment Organism
1,2-alpha-L-fucosidase
-
Bifidobacterium bifidum
AfcA from glycoside hydrolase family 95 Bifidobacterium bifidum