Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | no significant activation of the activity of crotonyl-CoA reductase is observed upon addition of either flavin adenine dinucleotide (0.018 and 0.072 mM) or flavin mononucleotide (0.013 and 0.130 mM) | Streptomyces collinus |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3)/pZYB3 cells | Streptomyces collinus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ammonium sulfate | enzyme activity is inhibited by ammonium sulfate ; however, this inhibition is overcome by addition of 10 mM guanidine | Streptomyces collinus | |
arachidoyl-CoA | 86% residual activity at 0.1 mM | Streptomyces collinus | |
butyryl-CoA | slight inhibition | Streptomyces collinus | |
Ca2+ | complete inhibition at 1 mM | Streptomyces collinus | |
Co2+ | complete inhibition at 1 mM | Streptomyces collinus | |
iodoacetamide | 40% inhibition at 1 mM | Streptomyces collinus | |
isomyristoyl-CoA | 78% residual activity at 0.1 mM | Streptomyces collinus | |
isopalmitoyl-CoA | 95% residual activity at 0.1 mM | Streptomyces collinus | |
Mg2+ | 30% inhibition at 1 mM | Streptomyces collinus | |
Mn2+ | complete inhibition at 1 mM | Streptomyces collinus | |
additional information | no significant inhibition of the activity of crotonyl-CoA reductase is observed upon addition of either flavin adenine dinucleotide (0.018 and 0.072 mM) or flavin mononucleotide (0.013 and 0.130 mM) | Streptomyces collinus | |
myristoyl-CoA | 36% residual activity at 0.1 mM | Streptomyces collinus | |
N-ethylmaleimide | 80% inhibition at 1 mM | Streptomyces collinus | |
NADP+ | - |
Streptomyces collinus | |
NADPH | concentrations of NADPH above 0.2 mM lead to inhibition of enzyme activity | Streptomyces collinus | |
p-chloromercuribenzoate | a 30-min incubation of crotonyl-CoA reductase with p-chloromercuribenzoate at 0.008 mM leads to approximately 8.5% inhibition of enzyme activity | Streptomyces collinus | |
palmitoyl-CoA | 24% residual activity at 0.1 mM | Streptomyces collinus | |
stearoyl-CoA | 92% residual activity at 0.1 mM | Streptomyces collinus | |
Zn2+ | 55% inhibition at 1 mM | Streptomyces collinus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
NADPH | in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 1 mM dithioerythritol and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.018 | - |
crotonyl-CoA | in 50 mM potassium phosphate, pH 7.5, 1 mM EDTA, 1 mM dithioerythritol and 10% (v/v) glycerol, at 30°C | Streptomyces collinus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
48000 | - |
2 * 48000, SDS-PAGE | Streptomyces collinus |
49400 | - |
2 * 49400, calculated from amino acid sequence | Streptomyces collinus |
85000 | - |
native protein, gel filtration | Streptomyces collinus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces collinus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
ammonium sulfate precipitation, DEAE-cellulose column chromatography, phenyl-Sepharose column chromatography, Mono Q column chromatography, Sephadex G-100 gel filtration, and phenyl-Superose gel filtration | Streptomyces collinus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.00091 | - |
native enzyme from crude extract, pH 7.5 at 30°C | Streptomyces collinus |
0.333 | - |
recombinant enzyme from crude extract, pH 7.5 at 30°C | Streptomyces collinus |
2.889 | - |
native enzyme after 3068fold purification, pH 7.5 at 30°C | Streptomyces collinus |
3.316 | - |
recombinant enzyme after 10fold purification, pH 7.5 at 30°C | Streptomyces collinus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
crotonyl-CoA + NADPH + H+ | the enzyme exhibits a high substrate specificity for crotonyl-CoA | Streptomyces collinus | butanoyl-CoA + NADP+ | - |
? | |
additional information | the enzyme is unable to catalyze the reduction of any other enoyl-CoA thioesters (acryloyl-CoA, trans-2-pentenoyl-CoA, trans-hexenoyl-CoA, trans-2-octenoyl-CoA, trans-2-dodecenoyl-CoA, trans-2-hexadecenoyl-CoA) or to utilize NADH as an electron donor. The enzyme is unable to reduce either the N-acetylcysteamine or the pantetheine thioester of crotonic acid | Streptomyces collinus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 48000, SDS-PAGE | Streptomyces collinus |
homodimer | 2 * 49400, calculated from amino acid sequence | Streptomyces collinus |
Synonyms | Comment | Organism |
---|---|---|
acyl-CoA:NADP+ trans-2-oxidoreductase | - |
Streptomyces collinus |
crotonyl-CoA reductase | - |
Streptomyces collinus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Streptomyces collinus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
at 40°C the enzyme retains 47% of its activity after 30 min | Streptomyces collinus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
- |
Streptomyces collinus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | NADPH is the sole electron donor for the reduction catalyzed by crotonyl-CoA reductase | Streptomyces collinus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0095 | - |
palmitoyl-CoA | in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.017 | - |
myristoyl-CoA | in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.4 | - |
isopalmitoyl-CoA | Ki above 0.4 mM, in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.5 | - |
isomyristoyl-CoA | in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.63 | - |
NADP+ | in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus | |
0.9 | - |
butyryl-CoA | in 50 mM Tris/HCI pH 6.5 and 10% (v/v) glycerol, at 30°C | Streptomyces collinus |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme plays a role in providing butyryl-CoA as a starter unit for straight-chain fatty acid biosynthesis | Streptomyces collinus |