Protein Variants | Comment | Organism |
---|---|---|
K173A | 37°C, t1/2 of the mutant enzyme is 60 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor | Thermoactinomyces intermedius |
K69A | 37°C, t1/2 of the mutant enzyme is 50 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor. Km-value for L-Phe is 1400fold higher compared to wild type enzyme, Km-value for phenylpyruvate is 128fold higher compared to wild type enzyme. Turnover number for deamination is 686fold lower than that of wild-type enzyme, turnover-number for amination is 43fold lower than that of wild-type enzyme | Thermoactinomyces intermedius |
K69A/K81A | 37°C, t1/2 of the mutant enzyme is 450 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor. Km-value for L-Phe is 200fold higher compared to wild type enzyme, Km-value for phenylpyruvate is 108fold higher compared to wild type enzyme. Turnover number for deamination is 110fold lower than that of wild-type enzyme, turnover-number for amination is 61fold lower than that of wild-type enzyme | Thermoactinomyces intermedius |
K81A | 37°C, t1/2 of the mutant enzyme is 38 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor. Turnover number for deamination is 440fold lower than that of wild-type enzyme, turnover-number for amination is 42fold lower than that of wild-type enzyme | Thermoactinomyces intermedius |
K89A | 37°C, t1/2 of the mutant enzyme is 75 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor | Thermoactinomyces intermedius |
K90A | 37°C, t1/2 of the mutant enzyme is 80 min, compared to 48 min for the wild type enzyme, without addition of substrate or cofactor | Thermoactinomyces intermedius |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
methyl acetyl phosphate | irreversible inactivation, simultaneous addition of substrate and coenzyme markedly protect from inactivation, the reagent can acetylate Lys69 and Lys81 | Thermoactinomyces intermedius |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
NADH | mutant enzyme K69A/K81A | Thermoactinomyces intermedius | |
0.026 | - |
NADH | mutant enzyme K81A | Thermoactinomyces intermedius | |
0.051 | - |
NADH | mutant enzyme K69A | Thermoactinomyces intermedius | |
0.052 | - |
phenylpyruvate | mutant enzyme K81A | Thermoactinomyces intermedius | |
0.057 | - |
NAD+ | mutant enzyme K81A | Thermoactinomyces intermedius | |
0.065 | - |
phenylpyruvate | wild-type enzyme | Thermoactinomyces intermedius | |
0.081 | - |
NAD+ | mutant enzyme K69A/K81A | Thermoactinomyces intermedius | |
0.083 | - |
NADH | wild-type enzyme | Thermoactinomyces intermedius | |
0.09 | - |
L-Phe | mutant enzyme K81A | Thermoactinomyces intermedius | |
0.1 | - |
L-Phe | wild-type enzyme | Thermoactinomyces intermedius | |
0.16 | - |
NAD+ | mutant enzyme K69A | Thermoactinomyces intermedius | |
0.17 | - |
NAD+ | wild-type enzyme | Thermoactinomyces intermedius | |
7 | - |
phenylpyruvate | mutant enzyme K69A/K81A | Thermoactinomyces intermedius | |
8.3 | - |
phenylpyruvate | mutant enzyme K69A | Thermoactinomyces intermedius | |
20 | - |
L-Phe | mutant enzyme K69A/K81A | Thermoactinomyces intermedius | |
140 | - |
L-Phe | mutant enzyme K69A | Thermoactinomyces intermedius |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoactinomyces intermedius | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-Phe + H2O + NAD+ | - |
Thermoactinomyces intermedius | phenylpyruvate + NH3 + NADH | - |
r |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
t1/2: 48 min, in absence of substrate or coenzyme. t1/2: 2000 min, in presence of 10 mM L-Leu and 1.0 mM NAD+ | Thermoactinomyces intermedius |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | turnover numbers of wild-type and mutant enzymes | Thermoactinomyces intermedius |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermoactinomyces intermedius | |
NADH | - |
Thermoactinomyces intermedius |