Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain M15 | Digitalis lanata |
DNA and amino acid sequence determination and analysis, sequence comparisons, expression of wild-type and mutant enzymes in Escherichia coli strain M15 | Arabidopsis thaliana |
Crystallization (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography | Digitalis lanata |
Protein Variants | Comment | Organism |
---|---|---|
C352G | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows reduced activity compared to the wild-type enzyme | Digitalis lanata |
D181T/L182Q | site-directed mutagenesis, mutation at motif V, the mutant shows reduced activity compared to the wild-type enzyme | Digitalis lanata |
F353M | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
F353P | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
G204N | site-directed mutagenesis, mutation at motif IV, the mutant shows reduced activity compared to the wild-type enzyme | Digitalis lanata |
L182Q | site-directed mutagenesis, mutation at motif V, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
M150L | site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
N205A | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme | Digitalis lanata |
N205M | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme | Digitalis lanata |
N205M/Y156V | site-directed mutagenesis, double mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme | Digitalis lanata |
R146T | site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
S248M | site-directed mutagenesis, mutation near the substrate binding site, inactive mutant | Digitalis lanata |
T65P | site-directed mutagenesis, mutation at near motif II, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
Y156V | site-directed mutagenesis, mutation at the substrate binding site, the mutant shows increased activity compared to the wild-type enzyme | Digitalis lanata |
Y302F | site-directed mutagenesis, mutation at motif IV, the mutant shows similar activity as the wild-type enzyme | Digitalis lanata |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.063 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
0.072 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata | |
0.085 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
0.09 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
0.116 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana | |
0.175 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
0.196 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
0.268 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana | |
0.333 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
0.334 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
0.362 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
1.15 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-cyclohexen-1-one + NADPH + H+ | Digitalis lanata | - |
cyclohexanone + NADP+ | - |
? | |
2-cyclohexen-1-one + NADPH + H+ | Arabidopsis thaliana | - |
cyclohexanone + NADP+ | - |
? | |
progesterone + NADPH + H+ | Digitalis lanata | - |
5beta-pregnane-3,20-dione + NADP+ | - |
? | |
progesterone + NADPH + H+ | Arabidopsis thaliana | - |
5beta-pregnane-3,20-dione + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | Q9STX2 | - |
- |
Digitalis lanata | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain M15 by nickel affinity chromatography | Arabidopsis thaliana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-cyclohexen-1-one + NADPH + H+ | - |
Digitalis lanata | cyclohexanone + NADP+ | - |
? | |
2-cyclohexen-1-one + NADPH + H+ | - |
Arabidopsis thaliana | cyclohexanone + NADP+ | - |
? | |
progesterone + NADPH + H+ | - |
Digitalis lanata | 5beta-pregnane-3,20-dione + NADP+ | - |
? | |
progesterone + NADPH + H+ | - |
Arabidopsis thaliana | 5beta-pregnane-3,20-dione + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
P5betaR | - |
Digitalis lanata |
P5betaR | - |
Arabidopsis thaliana |
progesterone 5beta-reductase | - |
Digitalis lanata |
progesterone 5beta-reductase | - |
Arabidopsis thaliana |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at | Digitalis lanata |
40 | - |
assay at | Arabidopsis thaliana |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0075 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata | |
0.011 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
0.019 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
0.03 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
0.037 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
0.17 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana | |
0.4 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
0.475 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
0.49 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
0.68 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata | |
1.1 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
1.11 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Digitalis lanata |
8 | - |
assay at | Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Digitalis lanata | |
NADPH | - |
Arabidopsis thaliana |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the aldoketo reductase (AKR) family, sequence comparisons, overview | Digitalis lanata |
evolution | the enzyme belongs to the aldoketo reductase (AKR) family, sequence comparisons, overview | Arabidopsis thaliana |
additional information | residues Tyr156, Asp205, and Ser248 are responsible for the low catalytic efficiency of the enzyme, substrate binding pocket structure, overview | Digitalis lanata |
additional information | the enzyme shows high catalytic efficiency | Arabidopsis thaliana |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.052 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
0.105 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata | |
0.122 | - |
progesterone | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
0.432 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
0.473 | - |
progesterone | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
0.588 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V | Digitalis lanata | |
0.628 | - |
progesterone | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana | |
1.19 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant Y156V/N205M | Digitalis lanata | |
1.464 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Digitalis lanata | |
2.425 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205M | Digitalis lanata | |
6.266 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant mutant N205A | Digitalis lanata | |
9.607 | - |
2-cyclohexen-1-one | pH 8.0, 40°C, recombinant wild-type enzyme | Arabidopsis thaliana |