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Literature summary for 1.1.1.35 extracted from
- (S)-3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA hydratase (FadB) from fatty acid degradation operon of Ralstonia eutropha H16 (2014), AMB Express, 4, 69.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis, genetic organization, sequence comparisons and phylogenetic tree of 3-hydroxyacyl-CoA dehydrogenases, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) | Cupriavidus necator |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| acetyl-CoA | inhibits the enzyme by 50% at 0.1 mM, further increase of inhibitor concentration reduces the activity of FadB' drastically | Cupriavidus necator |  |
| CoA | inhibits the enzyme by 20% at 0.1 mM, further increase of inhibitor concentration reduces the activity of FadB' drastically | Cupriavidus necator | |
| propionyl-CoA | - |
Cupriavidus necator | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.048 | - |
acetoacetyl-CoA | pH 6.5, 30°C, recombinant enzyme | Cupriavidus necator | |
| 0.076 | - |
acetoacetyl-CoA | pH 6.5, 30°C, recombinant enzyme in presence of 0.1 mM acetyl-CoA | Cupriavidus necator | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 85900 | - |
- |
Cupriavidus necator |
| 86000 | - |
- |
Cupriavidus necator |
| 87000 | - |
recombinant His-tagged enzyme, gel filtration | Cupriavidus necator |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-3-hydroxybutanoyl-CoA + NAD+ | Cupriavidus necator | - |
acetoacetyl-CoA + NADH + H+ | - |
r | |
| (S)-3-hydroxybutanoyl-CoA + NAD+ | Cupriavidus necator DSM 428 | - |
acetoacetyl-CoA + NADH + H+ | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Cupriavidus necator | Q0KEG0 | - |
- |
| Cupriavidus necator DSM 428 | Q0KEG0 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Cupriavidus necator |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1.5 | - |
purified recombinant enzyme, pH 6.5, 30°C, substrate (S)-3-hydroxybutanoyl-CoA with NADP+ | Cupriavidus necator |
| 16.5 | - |
purified recombinant enzyme, pH 6.5, 30°C, substrate (S)-3-hydroxybutanoyl-CoA with NAD+ | Cupriavidus necator |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-3-hydroxybutanoyl-CoA + NAD+ | - |
Cupriavidus necator | acetoacetyl-CoA + NADH + H+ | - |
r | |
| (S)-3-hydroxybutanoyl-CoA + NAD+ | enzyme FadB' converts (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA, while no conversion of (R)-3-hydroxybutyryl-CoA is detected | Cupriavidus necator | acetoacetyl-CoA + NADH + H+ | - |
r | |
| (S)-3-hydroxybutanoyl-CoA + NAD+ | - |
Cupriavidus necator DSM 428 | acetoacetyl-CoA + NADH + H+ | - |
r | |
| (S)-3-hydroxybutanoyl-CoA + NAD+ | enzyme FadB' converts (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA, while no conversion of (R)-3-hydroxybutyryl-CoA is detected | Cupriavidus necator DSM 428 | acetoacetyl-CoA + NADH + H+ | - |
r | |
| (S)-3-hydroxybutanoyl-CoA + NADP+ | relative activity of FadB' measured with NADP+ is less than 10% in comparison to the activity measured with NAD+ | Cupriavidus necator | acetoacetyl-CoA + NADPH + H+ | - |
? | |
| (S)-3-hydroxybutanoyl-CoA + NADP+ | relative activity of FadB' measured with NADP+ is less than 10% in comparison to the activity measured with NAD+ | Cupriavidus necator DSM 428 | acetoacetyl-CoA + NADPH + H+ | - |
? | |
| additional information | the N-terminal part of FadB' comprises an NAD+ binding site and is responsible for 3-hydroxyacyl-CoA dehydrogenase activity converting (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA. FadB' is strictly stereospecific to (S)-3-hydroxybutyryl-CoA and to prefers NAD+. NADP(H) is utilized at a rate of less than 10% in comparison to activity with NAD(H) | Cupriavidus necator | ? | - |
? | |
| additional information | the N-terminal part of FadB' comprises an NAD+ binding site and is responsible for 3-hydroxyacyl-CoA dehydrogenase activity converting (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA. FadB' is strictly stereospecific to (S)-3-hydroxybutyryl-CoA and to prefers NAD+. NADP(H) is utilized at a rate of less than 10% in comparison to activity with NAD(H) | Cupriavidus necator DSM 428 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 86000, recombinant His-tagged enzme, SDS-PAGE, 1 * 85900, about, sequence calculation | Cupriavidus necator |
| More | FadB' is a polypeptide comprising a N-terminal 3HCDH_N-domain, responsible for NAD+ binding, and a C-terminal enoyl-CoA hydratase domain, an with inverted domain order | Cupriavidus necator |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| (S)-3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA hydratase | - |
Cupriavidus necator |
| (S)-stereospecific 3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA hydratase | - |
Cupriavidus necator |
| FadB' | - |
Cupriavidus necator |
| H16_A0461 | - |
Cupriavidus necator |
| More | cf. EC 4.2.1.17 | Cupriavidus necator |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 35 | 40 | - |
Cupriavidus necator |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 7 | - |
Cupriavidus necator |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 10 | maximal activity at pH 6.0-7.0, 65% and 75% activity at pH 5.0 and pH 8.0, respectively, and 46% of the maximal activity at pH 10.0, and 26% of maximal activity at pH 4.0 | Cupriavidus necator |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | the enzyme prefers NAD+. NADP(H) is utilized at a rate of less than 10% in comparison to activity with NAD(H) | Cupriavidus necator | |
| NAD+ | - |
Cupriavidus necator | |
| NADH | - |
Cupriavidus necator | |
| NADP+ | - |
Cupriavidus necator | |
| NADPH | - |
Cupriavidus necator | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Cupriavidus necator | sequence calculation | - |
7.66 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | two (S)-3-hydroxyacyl-CoA dehydrogenase/enoyl-CoA hydratases, H16_A0461/FadB' and H16_A1526/FadB1, are involved in the FA degradation in Ralstonia eutropha H16. FadB' and FadB1 possess an enoyl-CoA hydratase activity, catalyzing hydrogenation of the unsaturated enoyl coenzyme A (CoA), and a 3-hydroxyacyl-CoA dehydrogenase activity, i.e. oxidation of the hydroxyl group into a keto group using one NAD+ molecule | Cupriavidus necator |
| metabolism | the enzyme is involved in fatty acid degradation metabolism | Cupriavidus necator |
| additional information | FadB' is an enzyme with two catalytic domains exhibiting a single monomeric structure and possessing a molecular weight of 86 kDa. The C-terminal part of the enzyme harbors enoyl-CoA hydratase activity, EC 4.2.1.17, and is able to convert trans-crotonyl-CoA to 3-hydroxybutyryl-CoA. The N-terminal part of FadB' comprises an NAD+ binding site and is responsible for 3-hydroxyacyl-CoA dehydrogenase activity converting (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA | Cupriavidus necator |
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Release 2023.1 (January 2023)
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