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Literature summary 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

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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.63 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

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.63 deoxyfuconojirimycin
-
 Bifidobacterium bifidum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.63 0.035
-
 2-fucosyllactose N421A mutant Bifidobacterium bifidum
3.2.1.63 0.056
-
 2-fucosyllactose N421G mutant Bifidobacterium bifidum
3.2.1.63 0.075
-
 2-fucosyllactose N423G mutant Bifidobacterium bifidum
3.2.1.63 0.089
-
 2-fucosyllactose D766A mutant Bifidobacterium bifidum
3.2.1.63 0.091
-
 2-fucosyllactose wild type Bifidobacterium bifidum
3.2.1.63 0.16
-
 2-fucosyllactose E566A mutant Bifidobacterium bifidum
3.2.1.63 0.19
-
 2-fucosyllactose D766E mutant Bifidobacterium bifidum
3.2.1.63 0.32
-
 2-fucosyllactose N423D mutant Bifidobacterium bifidum
3.2.1.63 17
-
 2-fucosyllactose E485A mutant Bifidobacterium bifidum

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.63 Bifidobacterium bifidum
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.63 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.63 2'-fucosyllactose + H2O
-
 Bifidobacterium bifidum fucose + lactose
-
 ?
3.2.1.63 methyl-2-alpha-L-fucopyranosyl-beta-D-galactoside + H2O
-
 Bifidobacterium bifidum L-fucose + methyl beta-D-galactoside
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.63 1,2-alpha-L-fucosidase
-
 Bifidobacterium bifidum
3.2.1.63 AfcA from glycoside hydrolase family 95 Bifidobacterium bifidum