Cloned (Comment) | Organism |
---|---|
expressed as GST-tagged enzyme in Escherichia coli BL21 cells | Solanum lycopersicum |
Protein Variants | Comment | Organism |
---|---|---|
C117A | the binding affinity of NADH in the mutant MDAR is lower than that of wild type MDAR as indicated by about 1.5-3fold increase of the Km value for NADH, the mutant is less thermo-stable than the wild type enzyme | Solanum lycopersicum |
C117S | the binding affinity of NADH in the mutant MDAR is lower than that of wild type MDAR as indicated by about 1.5-3fold increase of the Km value for NADH, the mutant is less thermo-stable than the wild type enzyme | Solanum lycopersicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00144 | - |
monodehydroascorbate | native wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.0015 | - |
monodehydroascorbate | recombinant wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.00151 | - |
monodehydroascorbate | mutant enzyme C117S, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.00155 | - |
monodehydroascorbate | mutant enzyme C117A, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.00621 | - |
NADH | recombinant wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.0063 | - |
NADH | native wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.00962 | - |
NADH | mutant enzyme C117S, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
0.01461 | - |
NADH | mutant enzyme C117A, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Solanum lycopersicum | 9507 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Solanum lycopersicum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
glutathione Sepharose column chromatography | Solanum lycopersicum |
Renatured (Comment) | Organism |
---|---|
using 0.8 M guanidine-HCl at 25°C for 24 h | Solanum lycopersicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
NADH + monodehydroascorbate | - |
Solanum lycopersicum | NAD+ + ascorbate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MDAR | - |
Solanum lycopersicum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | 62 | the activity of wild type MDAR has no much change after heat treatment at temperatures below 35°C, steep decrease of activity is observed between 35 and 55°C, and a complete loss in activity occurs above 62°C. The midpoint of thermal inactivation of wild type MDAR is about 52.1°C | Solanum lycopersicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
62.67 | - |
NADH | mutant enzyme C117A, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
89.89 | - |
NADH | mutant enzyme C117S, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
121.4 | - |
NADH | recombinant wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
126.6 | - |
NADH | native wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Solanum lycopersicum |
General Information | Comment | Organism |
---|---|---|
physiological function | MDARis a key enzyme in the ascorbate-glutathione cycle, MDAR is crucial for ascorbate regeneration and essential for maintaining the reduced pool of ascorbate | Solanum lycopersicum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4290 | - |
NADH | mutant enzyme C117A, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
9360 | - |
NADH | mutant enzyme C117S, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
19270 | - |
NADH | recombinant wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum | |
20420 | - |
NADH | native wild type enzyme, in 50 mM Tris-HCl (pH 7.5), at 25°C | Solanum lycopersicum |