EC Number | Cloned (Comment) | Organism |
---|---|---|
1.3.1.85 | expressed in Escherichia coli BL21 AI (DE3) cells | Kitasatospora setae |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.3.1.85 | apoenzyme and in complex with NADPH and (2S)-ethylmalonyl-CoA, sitting drop vapor diffusion method, using 100 mM Tris pH 8.0 and 20% (w/v) poly (acrylic acid sodium salt) 5100 | Kitasatospora setae |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.3.1.85 | E171A | the mutant shows reduced activity compared to the wild type enzyme | Kitasatospora setae |
1.3.1.85 | F170A | in the mutant, the carboxylation activity is decreased to 17% at the expense of increased reduction side reactivity compared to the wild type enzyme | Kitasatospora setae |
1.3.1.85 | F170Y | the mutant shows a slightly increased substrate inhibition, but otherwise very similar kinetic parameters as the wild type enzyme | Kitasatospora setae |
1.3.1.85 | H365N | the mutant shows a 20fold decreased activity compared with the wild type enzyme but still displays 93% of carboxylated product, even though the KM for CO2 is raised by more than 1 order of magnitude | Kitasatospora setae |
1.3.1.85 | N81L | the interaction of the enzyme with CO2 is severely disturbed by the N81L mutation. Stereospecificity of butyryl-CoA formation is almost completely lost in the mutant compared with the wild type enzyme | Kitasatospora setae |
1.3.1.85 | S119A | the mutant shows almost full carboxylation, but at more than 50fold reduced turnover frequency as compared to the wild type enzyme | Kitasatospora setae |
1.3.1.85 | T82D | the mutant shows almost full carboxylation, but at more than 50fold reduced turnover frequency as compared to the wild type enzyme | Kitasatospora setae |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.3.1.85 | 0.01 | - |
crotonyl-CoA | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.011 | - |
NADPH | mutant enzyme F170A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.0134 | - |
crotonyl-CoA | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.021 | - |
crotonyl-CoA | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.022 | - |
NADPH | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.0298 | - |
crotonyl-CoA | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.03 | - |
NADPH | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.03 | - |
CO2 | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.031 | - |
crotonyl-CoA | mutant enzyme F170A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.036 | - |
NADPH | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.036 | - |
crotonyl-CoA | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.037 | - |
NADPH | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.052 | - |
NADPH | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.08 | - |
CO2 | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.09 | - |
CO2 | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.112 | - |
NADPH | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.15 | - |
CO2 | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.155 | - |
CO2 | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.5 | - |
crotonyl-CoA | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 1.31 | - |
CO2 | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.3.1.85 | Kitasatospora setae | E4N096 | - |
- |
1.3.1.85 | Kitasatospora setae ATCC 33774 | E4N096 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.3.1.85 | HisTrap column chromatography | Kitasatospora setae |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.3.1.85 | crotonyl-CoA + CO2 + NADPH + H+ | - |
Kitasatospora setae | (2S)-ethylmalonyl-CoA + NADP+ | - |
? | |
1.3.1.85 | crotonyl-CoA + CO2 + NADPH + H+ | - |
Kitasatospora setae ATCC 33774 | (2S)-ethylmalonyl-CoA + NADP+ | - |
? | |
1.3.1.85 | additional information | the enzyme catalyzes the reduction of crotonyl-CoA to butyryl-CoA as a side reaction, but only in the absence of CO2 and at low catalytic efficiency | Kitasatospora setae | ? | - |
- |
|
1.3.1.85 | additional information | the enzyme catalyzes the reduction of crotonyl-CoA to butyryl-CoA as a side reaction, but only in the absence of CO2 and at low catalytic efficiency | Kitasatospora setae ATCC 33774 | ? | - |
- |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.3.1.85 | CCR | - |
Kitasatospora setae |
1.3.1.85 | ECR | - |
Kitasatospora setae |
1.3.1.85 | enoyl-CoA carboxylase/reductase | - |
Kitasatospora setae |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.3.1.85 | 0.157 | - |
NADPH | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.17 | - |
CO2 | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 0.172 | - |
crotonyl-CoA | mutant enzyme T82D, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 1.32 | - |
CO2 | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 1.47 | - |
NADPH | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 1.79 | - |
crotonyl-CoA | mutant enzyme S119A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 5 | - |
crotonyl-CoA | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 5.1 | - |
CO2 | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 5.1 | - |
crotonyl-CoA | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 6 | - |
NADPH | mutant enzyme E171A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 7.4 | - |
CO2 | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 8.1 | - |
NADPH | mutant enzyme H365N, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 8.3 | - |
crotonyl-CoA | mutant enzyme F170A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 11 | - |
NADPH | mutant enzyme F170A, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 56 | - |
NADPH | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 56 | - |
CO2 | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 78 | - |
CO2 | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 83 | - |
crotonyl-CoA | mutant enzyme F170Y, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 86 | - |
NADPH | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae | |
1.3.1.85 | 103 | - |
crotonyl-CoA | wild type enzyme, at pH 8.0 and 30°C | Kitasatospora setae |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.3.1.85 | NADPH | - |
Kitasatospora setae |