Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Ruminococcus albus |
Protein Variants | Comment | Organism |
---|---|---|
C485A | the mutant shows 43% of wild type activity with cellobiose. The mutant shows higher preference for D-glucosamine than the wild type enzyme | Ruminococcus albus |
E654A | inactive | Ruminococcus albus |
E654D | inactive | Ruminococcus albus |
Y648A | inactive | Ruminococcus albus |
Y648F | The mutant shows 23% of wild type activity with cellobiose. Apparent kcat/Km values of the mutant for D-mannose and 2-deoxy-D-glucose are 8.2 and 4.0fold higher than those of the wild type enzyme, respectively | Ruminococcus albus |
Y648V | the mutant shows 0.888% of wild type activity with cellobiose and has synthetic activity toward N-acetyl-D-glucosamine | Ruminococcus albus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.46 | - |
alpha-D-glucose 1-phosphate | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
1.68 | - |
cellobiose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
11.3 | - |
alpha-D-glucose 1-phosphate | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
11.8 | - |
D-mannose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
13.3 | - |
D-glucosamine | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
15.7 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
16.6 | - |
cellobiose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
17.4 | - |
N-acetyl-D-glucosamine | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
23.5 | - |
D-mannose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
30.7 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
31.4 | - |
cellobiose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
58.1 | - |
D-glucosamine | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
60.2 | - |
2-deoxy-D-glucose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
62.6 | - |
D-mannose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
83.1 | - |
2-deoxy-D-glucose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
164 | - |
2-deoxy-D-glucose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
200 | - |
D-mannose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-D-glucose 1-phosphate + D-glucose | Ruminococcus albus | - |
cellobiose + phosphate | - |
r | |
cellobiose + phosphate | Ruminococcus albus | - |
alpha-D-glucose 1-phosphate + D-glucose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ruminococcus albus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-chelating column chromatography, and gel filtration | Ruminococcus albus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose | - |
Ruminococcus albus | ? | - |
? | |
alpha-D-glucose 1-phosphate + D-glucosamine | - |
Ruminococcus albus | 4-O-beta-D-glucopyranosyl-D-glucosamine | - |
? | |
alpha-D-glucose 1-phosphate + D-glucose | - |
Ruminococcus albus | cellobiose + phosphate | - |
r | |
alpha-D-glucose 1-phosphate + D-mannose | - |
Ruminococcus albus | 4-O-beta-D-glucopyranosyl-D-mannose | - |
? | |
alpha-D-glucose 1-phosphate + N-acetyl-D-glucosamine | substrate for mutant Y648V only | Ruminococcus albus | 4-O-beta-D-glucopyranosyl-N-acetyl-D-glucosamine | - |
? | |
cellobiose + phosphate | - |
Ruminococcus albus | alpha-D-glucose 1-phosphate + D-glucose | - |
r |
Synonyms | Comment | Organism |
---|---|---|
Cbp | - |
Ruminococcus albus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
the enzyme is stable below 40°C | Ruminococcus albus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.626 | - |
D-mannose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
1.3 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
2.83 | - |
D-mannose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
5.27 | - |
D-mannose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
6.54 | - |
N-acetyl-D-glucosamine | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
7.83 | - |
2-deoxy-D-glucose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
9.56 | - |
D-glucosamine | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
21.5 | - |
2-deoxy-D-glucose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
35.4 | - |
D-glucosamine | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
58.3 | - |
D-mannose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
64.2 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
73.9 | - |
cellobiose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
79.1 | - |
cellobiose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
88.9 | - |
cellobiose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
92.3 | - |
alpha-D-glucose 1-phosphate | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
96.5 | - |
alpha-D-glucose 1-phosphate | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
119 | - |
2-deoxy-D-glucose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
- |
Ruminococcus albus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5.5 | 8.5 | the enzyme remains stable over the pH range of 5.5 to 8.5 | Ruminococcus albus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0198 | - |
2-deoxy-D-glucose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0263 | - |
D-mannose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0376 | - |
N-acetyl-D-glucosamine | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0477 | - |
2-deoxy-D-glucose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0532 | - |
D-mannose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0681 | - |
cellobiose | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
0.0829 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648V, at pH 6.0 and 37°C | Ruminococcus albus | |
0.112 | - |
D-mannose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
0.357 | - |
2-deoxy-D-glucose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
0.61 | - |
D-glucosamine | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
0.719 | - |
D-glucosamine | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
0.931 | - |
D-mannose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
1.43 | - |
2-deoxy-D-glucose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
2.09 | - |
alpha-D-glucose 1-phosphate | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
2.36 | - |
cellobiose | mutant enzyme Y648F, at pH 6.0 and 37°C | Ruminococcus albus | |
5.35 | - |
cellobiose | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
8.55 | - |
alpha-D-glucose 1-phosphate | mutant enzyme C485A, at pH 6.0 and 37°C | Ruminococcus albus | |
47.1 | - |
cellobiose | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus | |
63.2 | - |
alpha-D-glucose 1-phosphate | wild type enzyme, at pH 6.0 and 37°C | Ruminococcus albus |