EC Number | Application | Comment | Organism |
---|---|---|---|
1.1.1.35 | biofuel production | 3-hydroxybutyryl-CoA dehydrogenase is an enzyme involved in the synthesis of the biofuel n-butanol by converting acetoacetyl-CoA to 3-hydroxybutyryl-CoA, molecular mechanism of n-butanol biosynthesis, overview | Cupriavidus necator |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.35 | gene paaH1, recombinant expression of C-terminally His6-tagged wild-type or selenomethionine-labeled enzymes in Escherichia coli strain B834 | Cupriavidus necator |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.1.1.35 | purified recombinant enzyme in apoform, as selenomethionine-labeled enzyme, and in complex with substrates acetoacetyl-CoA and NAD+, hanging drop vapour diffusion method, mixing of 50 mg/ml wild-type protein or selenomethionine-labeled enzyme in 40 mM Tris-HCl, pH 8.0, 1 mM DTT, with reservoir solution containing 2 M ammonium sulfate, 0.1 M sodium cacodylate, pH 6.5, and 0.2 M sodium chloride, 22°C, 7 days, X-ray diffraction structure determination and analysis at 2.42-2.7 A resolution, molecular replacement using the crystal structure of the apo-form of RePaaH1, and structure modeling | Cupriavidus necator |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.35 | K56A | site-directed mutagenesis, the mutant shows about twofold increased activity compared to the wild-type enzyme | Cupriavidus necator |
1.1.1.35 | N190A | site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme | Cupriavidus necator |
1.1.1.35 | R52A | site-directed mutagenesis, the mutant shows highly decreased activity compared to the wild-type enzyme | Cupriavidus necator |
1.1.1.35 | S119A | site-directed mutagenesis, almost inactive mutant | Cupriavidus necator |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.35 | additional information | - |
additional information | Michaelis-Menten kinetics | Cupriavidus necator | |
1.1.1.35 | 0.01825 | - |
acetoacetyl-CoA | pH 8.0, 30°C, recombinant enzyme | Cupriavidus necator |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | Cupriavidus necator | - |
acetoacetyl-CoA + NADH + H+ | - |
r | |
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | Cupriavidus necator ATCC 17699 | - |
acetoacetyl-CoA + NADH + H+ | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.35 | Cupriavidus necator | Q0KEY8 | - |
- |
1.1.1.35 | Cupriavidus necator ATCC 17699 | Q0KEY8 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.35 | recombinant C-terminally His6-tagged wild-type or selenomethionine-labeled enzymes from Escherichia coli strain B834 by nickel affinity chromatography and gel filtration | Cupriavidus necator |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | - |
Cupriavidus necator | acetoacetyl-CoA + NADH + H+ | - |
r | |
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | substrates binding structure analysis, overview. The acetoacetyl-CoA substrate is positioned within the deep cleft between the N-terminal domain and C-terminal domain. The acetoacetyl moiety is positioned near the conserved catalytic residues Ser119, His140, and Asn190 | Cupriavidus necator | acetoacetyl-CoA + NADH + H+ | - |
r | |
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | - |
Cupriavidus necator ATCC 17699 | acetoacetyl-CoA + NADH + H+ | - |
r | |
1.1.1.35 | (S)-3-hydroxybutanoyl-CoA + NAD+ | substrates binding structure analysis, overview. The acetoacetyl-CoA substrate is positioned within the deep cleft between the N-terminal domain and C-terminal domain. The acetoacetyl moiety is positioned near the conserved catalytic residues Ser119, His140, and Asn190 | Cupriavidus necator ATCC 17699 | acetoacetyl-CoA + NADH + H+ | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.35 | (S)-3-hydroxybutyryl-CoA dehydrogenase | - |
Cupriavidus necator |
1.1.1.35 | PaaH1 | - |
Cupriavidus necator |
1.1.1.35 | RePaaH1 | - |
Cupriavidus necator |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.35 | 30 | - |
assay at | Cupriavidus necator |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.35 | 45500 | - |
acetoacetyl-CoA | pH 8.0, 30°C, recombinant enzyme | Cupriavidus necator |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.35 | 8 | - |
assay at | Cupriavidus necator |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.35 | NAD+ | the NAD+ cofactor is bound to the N-terminal Rossmann fold, the NAD+-binding pocket is made up of 5 loops, enzyme binding structure analysis, overview | Cupriavidus necator | |
1.1.1.35 | NADH | - |
Cupriavidus necator |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.1.1.35 | additional information | conserved catalytic residues are Ser119, His140, and Asn190 | Cupriavidus necator |