Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli strain TG1 | Peptoniphilus asaccharolyticus |
Protein Variants | Comment | Organism |
---|---|---|
D245K | the mutation shows reduced activity and 36.4fold discrimination against NADPH compared to NADH | Peptoniphilus asaccharolyticus |
E243D | the mutation shows reduced activity and 130fold discrimination against NADPH compared to NADH | Peptoniphilus asaccharolyticus |
E243K | the mutation shows reduced activity and almost no discrimination against NADPH compared to NADH | Peptoniphilus asaccharolyticus |
E243R | the mutation shows reduced activity and almost no discrimination against NADPH compared to NADH | Peptoniphilus asaccharolyticus |
W244S | the mutation shows reduced activity and 205fold discrimination against NADPH compared to NADH | Peptoniphilus asaccharolyticus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0044 | - |
NADH | wild type enzyme, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.0076 | - |
NADH | mutant enzyme W244S, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.0123 | - |
NADH | mutant enzyme E243D, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.018 | - |
NADH | mutant enzyme D245K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.036 | - |
NADH | mutant enzyme E243K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.0384 | - |
NADPH | mutant enzyme E243R, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.0385 | - |
NADH | mutant enzyme E243R, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.053 | - |
NADPH | mutant enzyme E243K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.238 | - |
NADPH | mutant enzyme D245K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.431 | - |
NADPH | mutant enzyme W244S, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
0.476 | - |
NADPH | mutant enzyme E243D, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
1.64 | - |
NADPH | wild type enzyme, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Peptoniphilus asaccharolyticus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Q sepharose column chromatography | Peptoniphilus asaccharolyticus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.29 | - |
mutant enzyme W244S, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
0.32 | - |
wild type enzyme, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
0.33 | - |
mutant enzyme W244S, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
0.33 | - |
wild type enzyme, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
0.34 | - |
mutant enzyme W244S, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
0.5 | - |
mutant enzyme W244S, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
0.51 | - |
mutant enzyme W244S, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
0.73 | - |
wild type enzyme, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
0.87 | - |
mutant enzyme D245K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
0.9 | - |
mutant enzyme E243R, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
1.12 | - |
mutant enzyme D245K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
1.14 | - |
mutant enzyme D245K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
1.23 | - |
wild type enzyme, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
1.32 | - |
wild type enzyme, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
1.47 | - |
mutant enzyme E243D, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
1.47 | - |
mutant enzyme E243D, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
1.53 | - |
mutant enzyme D245K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
1.83 | - |
mutant enzyme E243R, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
2.16 | - |
mutant enzyme D245K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
2.26 | - |
mutant enzyme E243R, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
2.3 | - |
mutant enzyme E243R, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
2.98 | - |
mutant enzyme E243D, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
3 | - |
mutant enzyme E243R, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
3.05 | - |
mutant enzyme E243R, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
3.27 | - |
mutant enzyme E243K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
3.41 | - |
mutant enzyme E243R, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
3.82 | - |
mutant enzyme W244S, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
4.1 | - |
mutant enzyme E243R, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
4.93 | - |
mutant enzyme E243D, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
5 | - |
mutant enzyme E243D, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
5.2 | - |
mutant enzyme E243K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
5.8 | - |
mutant enzyme D245K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
6.78 | - |
mutant enzyme W244S, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
7.2 | - |
mutant enzyme E243D, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
9.01 | - |
mutant enzyme E243K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
9.6 | - |
mutant enzyme E243K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
9.8 | - |
wild type enzyme, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.0 | Peptoniphilus asaccharolyticus |
11 | - |
mutant enzyme E243R, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
12 | - |
mutant enzyme E243R, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
12.5 | - |
mutant enzyme D245K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
15 | - |
mutant enzyme E243D, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
17.6 | - |
mutant enzyme E243K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
19 | - |
mutant enzyme E243K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
20 | - |
mutant enzyme W244S, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
20.1 | - |
mutant enzyme E243K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
20.6 | - |
wild type enzyme, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 6.5 | Peptoniphilus asaccharolyticus |
25.3 | - |
mutant enzyme E243K, with 0.1 mM NADPH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
30 | - |
mutant enzyme D245K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
34 | - |
mutant enzyme E243D, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
50 | - |
wild type enzyme, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.0 | Peptoniphilus asaccharolyticus |
54 | - |
mutant enzyme E243K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
56 | - |
mutant enzyme E243K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
56.5 | - |
mutant enzyme W244S, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
62 | - |
mutant enzyme W244S, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
68 | - |
mutant enzyme D245K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
90 | - |
mutant enzyme E243D, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
101.8 | - |
mutant enzyme D245K, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
102 | - |
mutant enzyme E243D, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
155 | - |
wild type enzyme, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 7.5 | Peptoniphilus asaccharolyticus |
174 | - |
wild type enzyme, with 0.1 mM NADH, 20 mM oxoglutarate and 100 mM ammonium chloride at pH 8.0 | Peptoniphilus asaccharolyticus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | - |
Peptoniphilus asaccharolyticus | 2-oxoglutarate + NH3 + NADH + H+ | - |
? | |
L-glutamate + H2O + NADP+ | weak reaction | Peptoniphilus asaccharolyticus | 2-oxoglutarate + NH3 + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GDH | - |
Peptoniphilus asaccharolyticus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.67 | - |
NADPH | mutant enzyme E243R, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
1.71 | - |
NADH | mutant enzyme E243R, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
3 | - |
NADPH | mutant enzyme W244S, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
5.06 | - |
NADPH | mutant enzyme E243D, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
6 | - |
NADPH | mutant enzyme D245K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
10 | - |
NADPH | wild type enzyme, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
10.6 | - |
NADH | mutant enzyme E243K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
10.9 | - |
NADH | mutant enzyme W244S, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
14.8 | - |
NADPH | mutant enzyme E243K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
16.4 | - |
NADH | mutant enzyme D245K, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
16.9 | - |
NADH | mutant enzyme E243D, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus | |
31.3 | - |
NADH | wild type enzyme, in 100 mM potassium phosphate buffer at pH 7 with 20 mM oxoglutarate and 100 mM ammonium chloride | Peptoniphilus asaccharolyticus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 8 | - |
Peptoniphilus asaccharolyticus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | strong preference (1170fold) for NADH over NADPH | Peptoniphilus asaccharolyticus |