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Literature summary for 1.20.1.1 extracted from

  • Woodyer, R.; van der Donk, W.A.; Zhao, H.
    Optimizing a biocatalyst for improved NAD(P)H regeneration: directed evolution of phosphite dehydrogenase (2006), Comb. Chem. High Throughput Screen., 9, 237-245.
    View publication on PubMed
Search for more literature for 1.20.1.1

Application

Application Comment Organism
biotechnology improvement of enzyme for use in regeneration of NADH and NADPH, application in bioconversion of trimethylpyruvate to L-tert-leucine as model reaction Pseudomonas stutzeri

Protein Variants

Protein Variants Comment Organism
D13E/M26I/E175A/E332N/C336D mutant obtained by directed evolution, round 4 Pseudomonas stutzeri
D13E/M26I/E175A/T181S/A308T/E332N/C336D mutant obtained by directed evolution, round 6, strong decrease in KM value for NADP compared to wild-type Pseudomonas stutzeri
D13E/M26I/E175A/T181S/E332N/C336D mutant obtained by directed evolution, round 5 Pseudomonas stutzeri
D13E/M26I/E332N/C336D mutant obtained by directed evolution, round 3 Pseudomonas stutzeri
E175A mutant with significantly increased kcat value for NADP+ Pseudomonas stutzeri
K330* mutant obtained by directed evolution, round 1 Pseudomonas stutzeri
M26I/E332N/C336D mutant obtained by directed evolution, round 2 Pseudomonas stutzeri

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.014
-
 phosphite cosubstrate NAD+, mutant K330*, pH 7.3, 25°C Pseudomonas stutzeri
0.027
-
 phosphite cosubstrate NAD+, mutant M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.03
-
 phosphite cosubstrate NAD+, mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.032
-
 NAD+ mutant M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.034
-
 NAD+ mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.037
-
 NAD+ mutant K330*, pH 7.3, 25°C Pseudomonas stutzeri
0.039
-
 NAD+ mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.041
-
 NAD+ mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.045
-
 NAD+ mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.047
-
 phosphite cosubstrate NAD+, wild-type, pH 7.3, 25°C Pseudomonas stutzeri
0.048
-
 phosphite cosubstrate NAD+, mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.053
-
 NAD+ wild-type, pH 7.3, 25°C Pseudomonas stutzeri
0.056
-
 NADP+ mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.057
-
 NADP+ mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.062
-
 phosphite cosubstrate NADP+, mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.065
-
 NADP+ mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.07
-
 phosphite cosubstrate NADP+, mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.082
-
 phosphite cosubstrate NAD+, mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.084
-
 phosphite cosubstrate NAD+, mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.087
-
 phosphite cosubstrate NADP+, mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
0.138
-
 phosphite cosubstrate NADP+, mutant E175A, pH 7.3, 25°C Pseudomonas stutzeri
0.14
-
 NADP+ mutant E175A, pH 7.3, 25°C Pseudomonas stutzeri
1.07
-
 NADP+ mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
1.9
-
 phosphite cosubstrate NADP+, mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
1.9
-
 phosphite cosubstrate NADP+, wild-type, pH 7.3, 25°C Pseudomonas stutzeri
2.5
-
 NADP+ wild-type, pH 7.3, 25°C Pseudomonas stutzeri

Organism

Organism UniProt Comment Textmining
Pseudomonas stutzeri
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
phosphite + H2O + NAD+
-
 Pseudomonas stutzeri phosphate + NADH + H+
-
 ?
phosphite + H2O + NADP+
-
 Pseudomonas stutzeri phosphate + NADPH + H+
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4
-
 NADP+ wild-type, pH 7.3, 25°C Pseudomonas stutzeri
1.75
-
 NADP+ mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
1.8
-
 NADP+ mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
1.8
-
 NADP+ mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
1.9
-
 NADP+ mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
2.2
-
 NADP+ mutant E175A, pH 7.3, 25°C Pseudomonas stutzeri
2.8
-
 NAD+ mutant K330*, pH 7.3, 25°C Pseudomonas stutzeri
2.9
-
 NAD+ wild-type, pH 7.3, 25°C Pseudomonas stutzeri
3.6
-
 NAD+ mutant M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
3.7
-
 NAD+ mutant D13E/M26I/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
4
-
 NAD+ mutant D13E/M26I/E175A/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
4.3
-
 NAD+ mutant D13E/M26I/E175A/T181S/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri
5.7
-
 NAD+ mutant D13E/M26I/E175A/T181S/A308T/E332N/C336D, pH 7.3, 25°C Pseudomonas stutzeri