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Literature summary for 1.4.1.2 extracted from

  • Carrigan, J.B.; Engel, P.C.
    Probing the determinants of coenzyme specificity in Peptostreptococcus asaccharolyticus glutamate dehydrogenase by site-directed mutagenesis (2007), FEBS J., 274, 5167-5174.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain TG1 Peptoniphilus asaccharolyticus

Protein Variants

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 [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

Organism UniProt Comment Textmining
Peptoniphilus asaccharolyticus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Q sepharose column chromatography Peptoniphilus asaccharolyticus

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

Synonyms Comment Organism
GDH
-
Peptoniphilus asaccharolyticus

Turnover Number [1/s]

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 Range

pH Minimum pH Maximum Comment Organism
6 8
-
Peptoniphilus asaccharolyticus

Cofactor

Cofactor Comment Organism Structure
NADH strong preference (1170fold) for NADH over NADPH Peptoniphilus asaccharolyticus