Protein Variants | Comment | Organism |
---|---|---|
R100K | cob(II)alamin accumulates to a concentration similar to that of the wild-type enzyme, homolysis of the coenzyme is slower by an order of magnitude, compared to wild-type enzyme, glutamate binding is significantly weakened. Mutant does not exhibit the very large deuterium isotope effects that are observed for homolysis of the coenzyme when the wild-type enzyme is reacted with deuterated substrates. Km-value for glutamate is reduced 121fold compared to wild-type enzyme, KM-value for glutamate is increased 17fold compared to wild-type enzyme | Clostridium cochlearium |
R100M | no cob(II)alamin detected in UV-visible spectrum. Km-value for glutamate is reduced 276fold compared to wild-type enzyme, KM-value for glutamate is increased 13fold compared to wild-type enzyme | Clostridium cochlearium |
R100Y | no cob(II)alamin detected in UV-visible spectrum. Km-value for glutamate is reduced 322fold compared to wild-type enzyme, KM-value for glutamate is increased 17fold compared to wild-type enzyme | Clostridium cochlearium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.58 | - |
L-glutamate | 25°C, wild-type enzyme | Clostridium cochlearium | |
7.6 | - |
L-glutamate | 25°C, mutant enzyme R100Y | Clostridium cochlearium | |
10 | - |
L-glutamate | 25°C, mutant enzyme R100K | Clostridium cochlearium | |
13 | - |
L-glutamate | 25°C, mutant enzyme R100M | Clostridium cochlearium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium cochlearium | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | - |
Clostridium cochlearium | L-threo-3-methylaspartate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0181 | - |
L-glutamate | 25°C, mutant enzyme R100Y | Clostridium cochlearium | |
0.0211 | - |
L-glutamate | 25°C, mutant enzyme R100M | Clostridium cochlearium | |
0.048 | - |
L-glutamate | 25°C, mutant enzyme R100K | Clostridium cochlearium | |
5.8 | - |
L-glutamate | 25°C, wild-type enzyme | Clostridium cochlearium |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
adenosylcobalamin | Km-value for wild-type enzyme is 0.005 mM. No cob(II)alamin detected in UV-visible spectrum of mutant enzymes R100M and R100Y. In mutant enzyme R100K cob(II)alamin accumulates to a concentration similar to that of the wild-type enzyme, homolysis of the coenzyme is slower by an order of magnitude, compared to wild-type enzyme. Mutant does not exhibit the very large deuterium isotope effects that are observed for homolysis of the coenzyme when the wild-type enzyme is reacted with deuterated substrates | Clostridium cochlearium |