Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.1.211 extracted from

  • Nishimoto, M.; Hidaka, M.; Nakajima, M.; Fushinobu, S.; Kitaoka, M.
    Identification of amino acid residues that determine the substrate preference of 1,3-beta-galactosyl-N-acetylhexosamine phosphorylase (2012), J. Mol. Catal. B, 74, 97-102.
No PubMed abstract available
Search for more literature for 2.4.1.211

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis, exxpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) Bifidobacterium longum

Protein Variants

Protein Variants Comment Organism
P161S site-directed mutagenesis, the mutation leads to an increase in the selectivity on lacto-N-biose I Bifidobacterium longum
P161S/S336A site-directed mutagenesis Bifidobacterium longum
S336A site-directed mutagenesis, the mutation leads to an increase in the selectivity on lacto-N-biose I Bifidobacterium longum
V162T site-directed mutagenesis, the mutation leads to an increase in the selectivity on galacto-N-biose Bifidobacterium longum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.8
-
 lacto-N-biose I pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum
6.7
-
 galacto-N-biose pH 7.5, 30°C, mutant V162T Bifidobacterium longum
7.7
-
 galacto-N-biose pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum
8.4
-
 lacto-N-biose I pH 7.5, 30°C, mutant P161S Bifidobacterium longum
13
-
 lacto-N-biose I pH 7.5, 30°C, mutant S336A Bifidobacterium longum
20
-
 galacto-N-biose pH 7.5, 30°C, mutant S336A Bifidobacterium longum
22
-
 lacto-N-biose I pH 7.5, 30°C, mutant V162T Bifidobacterium longum
27
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S Bifidobacterium longum
46
-
 lacto-N-biose I pH 7.5, 30°C, mutant P161S/S336A Bifidobacterium longum
120
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S/S336A Bifidobacterium longum

Organism

Organism UniProt Comment Textmining
Bifidobacterium longum E8MF13
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography Bifidobacterium longum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
galacto-N-biose + phosphate
-
 Bifidobacterium longum alpha-D-galactose 1-phosphate + N-acetyl-D-galactosamine
-
 ?
lacto-N-biose I + phosphate
-
 Bifidobacterium longum alpha-D-galactose 1-phosphate + N-acetyl-D-glucosamine
-
 ?
additional information substrate specificity, overview. Structurally determined substrate preference for galacto-N-biose and lacto-N-biose I Bifidobacterium longum ?
-
 ?

Synonyms

Synonyms Comment Organism
GalHexNAcP
-
 Bifidobacterium longum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Bifidobacterium longum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5
-
 galacto-N-biose pH 7.5, 30°C, mutant S336A Bifidobacterium longum
3.7
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S/S336A Bifidobacterium longum
15
-
 lacto-N-biose I pH 7.5, 30°C, mutants S336A and Bifidobacterium longum
27
-
 lacto-N-biose I pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum
27
-
 galacto-N-biose pH 7.5, 30°C, mutant V162T Bifidobacterium longum
33
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S Bifidobacterium longum
33
-
 lacto-N-biose I pH 7.5, 30°C, mutant V162T Bifidobacterium longum
39
-
 lacto-N-biose I pH 7.5, 30°C, mutant P161S Bifidobacterium longum
65
-
 galacto-N-biose pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Bifidobacterium longum

General Information

General Information Comment Organism
evolution the GalHexNAcP belongs to the glycoside hydrolase family 112, GH112 Bifidobacterium longum
additional information the residues at positions 162, 161, and 336 determine the substrate specificity, structure-function relationship, molecular docking, and specificity prediction, overview. The side-chain hydroxyl group of S336 forms a hydrogen bond with the side-chain nitrogen atom of R358, which plays a significant role in catalysis Bifidobacterium longum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.031
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S/S336A Bifidobacterium longum
0.17
-
 galacto-N-biose pH 7.5, 30°C, mutant S336A Bifidobacterium longum
0.34
-
 lacto-N-biose I pH 7.5, 30°C, mutant P161S/S336A Bifidobacterium longum
1.1
-
 lacto-N-biose I pH 7.5, 30°C, mutant S336A Bifidobacterium longum
1.2
-
 galacto-N-biose pH 7.5, 30°C, mutant P161S Bifidobacterium longum
1.5
-
 lacto-N-biose I pH 7.5, 30°C, mutant V162T Bifidobacterium longum
4
-
 galacto-N-biose pH 7.5, 30°C, mutant V162T Bifidobacterium longum
4.6
-
 lacto-N-biose I pH 7.5, 30°C, mutant P161S Bifidobacterium longum
8.5
-
 galacto-N-biose pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum
9.3
-
 lacto-N-biose I pH 7.5, 30°C, wild-type enzyme Bifidobacterium longum