Cloned (Comment) | Organism |
---|---|
overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Comamonas testosteroni |
Protein Variants | Comment | Organism |
---|---|---|
K159A | site-directed mutagenesis, the mutant shows altered kinetics and pH profile, and 200fold reduced activity compared to the wild-type | Comamonas testosteroni |
S114A | site-directed mutagenesis, the mutant shows altered kinetics and pH profile, and 3400fold reduced activity compared to the wild-type | Comamonas testosteroni |
S114A/Y155F | site-directed mutagenesis, the mutant shows altered kinetics and pH profile, and 200000fold reduced activity compared to the wild-type | Comamonas testosteroni |
Y155F | site-directed mutagenesis, the mutant shows altered kinetics and pH profile, and 2800fold reduced activity compared to the wild-type | Comamonas testosteroni |
Y155F/K159A | site-directed mutagenesis, the mutant shows altered kinetics and pH profile, and 9400fold reduced activity compared to the wild-type | Comamonas testosteroni |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics | Comamonas testosteroni | |
0.00044 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant S114A/Y155F | Comamonas testosteroni | |
0.0055 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged wild-type | Comamonas testosteroni | |
0.007 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant S114A | Comamonas testosteroni | |
0.065 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant K159A | Comamonas testosteroni | |
0.074 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant Y155F | Comamonas testosteroni | |
0.1 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant Y155F/K159A | Comamonas testosteroni |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
70000 | - |
about, recombinant His-tagged wild-type and mutant enzymes, gel filtration | Comamonas testosteroni |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
androsterone + NAD+ | Comamonas testosteroni | - |
5alpha-androstane-3,17-dione + NADH | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Comamonas testosteroni | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, and gel filtration to homogeneity | Comamonas testosteroni |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ | the catalytic triad is composed of Ser-114, Tyr-155, and Lys-159, structural analysis and sequence alignment of the short chain dehydrogenase/reductase family | Comamonas testosteroni |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Comamonas testosteroni |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
androsterone + NAD+ | - |
Comamonas testosteroni | 5alpha-androstane-3,17-dione + NADH | - |
r | |
additional information | bifunctional enzyme also catalyzing the reaction of carbonyl reductase, EC 1.1.1.184 | Comamonas testosteroni | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 28349-28425, recombinant His-tagged wild-type and mutant enzymes, mass spectrometry | Comamonas testosteroni |
Synonyms | Comment | Organism |
---|---|---|
3alpha-HSD/CR | - |
Comamonas testosteroni |
3alpha-hydroxysteroid dehydrogenase/carbonyl reductase | - |
Comamonas testosteroni |
More | cf. EC 1.1.1.184 | Comamonas testosteroni |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Comamonas testosteroni |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0028 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant S114A/Y155F | Comamonas testosteroni | |
0.003 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant Y155F/K159A | Comamonas testosteroni | |
0.12 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant S114A | Comamonas testosteroni | |
1.5 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant Y155F | Comamonas testosteroni | |
18.9 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged mutant K159A | Comamonas testosteroni | |
318 | - |
androsterone | pH 9.0, 25°C, recombinant His-tagged wild-type | Comamonas testosteroni |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Comamonas testosteroni |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 11 | pH profile, recombinant His-tagged wild-type and mutant enzymes | Comamonas testosteroni |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Comamonas testosteroni | |
NADH | - |
Comamonas testosteroni |