Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) as soluble N-terminally His-tagged enzymes | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
S137A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Rattus norvegicus |
S137C | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Rattus norvegicus |
S137T | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137C | Rattus norvegicus | |
0.044 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged wild-type enzyme | Rattus norvegicus | |
0.184 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137A | Rattus norvegicus | |
0.263 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137T | Rattus norvegicus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Rattus norvegicus | 5739 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-oxoacyl-CoA + NADH | Rattus norvegicus | key enzyme involved in fatty acid oxidation | 3-hydroxyacyl-CoA + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography | Rattus norvegicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-3-hydroxyacyl-CoA + NAD+ = 3-oxoacyl-CoA + NADH + H+ | highly conserved residue Ser137 is essential for activity | Rattus norvegicus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
the N-terminal His-tag does notinfluence enzyme activity | Rattus norvegicus |
452 | - |
purified recombinant His-tagged wild-type enzyme | Rattus norvegicus |
Storage Stability | Organism |
---|---|
-80°C, purified recombinant His-tagged wild-type and mutant enzymes, stable for at least 3 months | Rattus norvegicus |
4°C, purified recombinant His-tagged wild-type enzyme, 1 week, no significant loss of activity | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3-oxoacyl-CoA + NADH | key enzyme involved in fatty acid oxidation | Rattus norvegicus | 3-hydroxyacyl-CoA + NAD+ | - |
r | |
acetoacetyl-CoA + NADH + H+ | - |
Rattus norvegicus | 3-hydroxybutyryl-CoA + NAD+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
short-chain 3-hydroxyacyl-CoA dehydrogenase | - |
Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.019 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137C | Rattus norvegicus | |
0.026 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137T | Rattus norvegicus | |
0.12 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged mutant S137A | Rattus norvegicus | |
569 | - |
acetoacetyl-CoA | pH 5.0, recombinant His-tagged wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
phosphate buffer | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Rattus norvegicus | |
NADH | - |
Rattus norvegicus |