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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Bifidobacterium longum | E8MF13 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography | Bifidobacterium longum |
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 | Comment | Organism |
---|---|---|
GalHexNAcP | - |
Bifidobacterium longum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bifidobacterium longum |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bifidobacterium longum |
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 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 |