Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Comamonas testosteroni |
Protein Variants | Comment | Organism |
---|---|---|
K159A | site-directed mutagenesis, the mutation changes the rate-limiting step to the hydride transfer, proton transfer is blocked in the mutant but can be rescued using exogenous proton acceptors, such as buffers, small primary amines, and azide, overview | Comamonas testosteroni |
K159M | site-directed mutagenesis, the mutation changes the rate-limiting step to the hydride transfer, proton transfer is blocked in the mutant but can be rescued using exogenous proton acceptors, such as buffers, small primary amines, and azide, overview | Comamonas testosteroni |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes in presence or absence of CAPS and methylamine, overview | Comamonas testosteroni | |
0.0051 | - |
androsterone | pH 10.4, 25°C, recombinant wild-type enzyme | Comamonas testosteroni | |
0.033 | - |
androsterone | pH 10.4, 25°C, recombinant mutant K159A | Comamonas testosteroni |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
androsterone + NAD+ | Comamonas testosteroni | - |
androstanedione + NADH | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Comamonas testosteroni | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes ifrom Escherichia coli strain BL21(DE3) | Comamonas testosteroni |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
a 3alpha-hydroxysteroid + NAD(P)+ = a 3-oxosteroid + NAD(P)H + H+ | The overall oxidoreductive reaction comprises the deprotonation of tyrosine, proton abstraction by the tyrosinate anion, and hydride transfer from the hydroxysteroid to NAD+, followed by the release of a proton from the hydroxy group of tyrosine to the solution., The proton transfer to the external base, from Tyr155 to Lys159 via 2'-OH of ribose, with a late transition state is the rate-limiting step, mechanism, overview | 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 | androstanedione + NADH | - |
r | |
androsterone + NAD+ | the intramolecular proton transfer is a rate-limiting step, with a concomitant releasing of protons to bulk solvent | Comamonas testosteroni | androstanedione + NADH | - |
r |
Synonyms | Comment | Organism |
---|---|---|
3alpha-HSD/CR | - |
Comamonas testosteroni |
3alpha-hydroxysteroid dehydrogenase/carbonyl reductase | - |
Comamonas testosteroni |
More | cf. EC 1.1.1.50 | Comamonas testosteroni |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Comamonas testosteroni |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10.4 | - |
assay at | Comamonas testosteroni |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Comamonas testosteroni | |
NADH | - |
Comamonas testosteroni |