Cloned (Comment) | Organism |
---|---|
a heterologous expression platform is established for the three structural genes of S25DH subunits together with an essential chaperone in the denitrifying betaproteobacterium Thauera aromatica K172. Using this system, steroid C-25 dehydrogenase S25DH1 and three isoenzymes (S25DH2, S25DH3, and S25DH4) are overproduced in a soluble, active form allowing a straightforward purification of nontagged alphabetagamma complexes | Sterolibacterium denitrificans |
General Stability | Organism |
---|---|
in the presence of 5 mM K3[Fe(CN)6], activity expressed in Thauera aromatica crude extracts is stable in air at 30°C for 5 to 8 h but decreased to 50% after 24 h | Sterolibacterium denitrificans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
beta-sitost-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.123 | - |
7-dehydrocholesterol | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.124 | - |
cholest-4-en-3-one | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.29 | - |
vitamin D3 | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.34 | - |
campest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.39 | - |
cholest-4-en-3-one | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.45 | - |
cholest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
1.84 | - |
campest-4-en-3-one | isoenzyme S25DH3, pH 7.0, 30°C | Sterolibacterium denitrificans |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mo2+ | the enzyme contains molybdenum | Sterolibacterium denitrificans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
160000 | - |
gel filtration, isoenzyme S25DH4 | Sterolibacterium denitrificans |
167000 | - |
gel filtration, isoenzyme S25DH, isoenzyme S25DH2, isoenzyme S25DH4 | Sterolibacterium denitrificans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cholest-4-en-3-one + acceptor + H2O | Sterolibacterium denitrificans | - |
25-hydroxycholest-4-en-3-one + reduced acceptor | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sterolibacterium denitrificans | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Sterolibacterium denitrificans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
7-dehydrocholesterol + acceptor + H2O | high activity with isoenzyme S25DH2, low activity with isoenzyme S25DH1. No activity with isoenzyme S25DH1, S25DH2 and S25DH3 | Sterolibacterium denitrificans | 25-hydroxy-7-dehydrocholesterol + reduced acceptor | - |
? | |
beta-sitost-4-en-3-one + acceptor + H2O | high activity with isoenzyme S25DH4. No activity with isoenzyme S25DH1 and S25DH2 | Sterolibacterium denitrificans | 25-hydroxysitost-4-en-3-one + reduced acceptor | - |
? | |
campest-4-en-3-one + acceptor + H2O | high activity with isoenzyme S25DH4, low activity with isoenzyme S25DH3. No activity with isoenzyme S25DH1 and S25DH2 | Sterolibacterium denitrificans | 25-hydroxycampest-4-en-3-one + reduced acceptor | - |
? | |
cholest-4-en-3-one + acceptor + H2O | - |
Sterolibacterium denitrificans | 25-hydroxycholest-4-en-3-one + reduced acceptor | - |
? | |
cholest-4-en-3-one + acceptor + H2O | high activity with isoenzymes S25DH1, S25DH2 and S25DH1. No activity with isoenzyme S25DH3 | Sterolibacterium denitrificans | 25-hydroxycholest-4-en-3-one + reduced acceptor | - |
? | |
vitamin D3 + acceptor + H2O | high activity with isoenzyme S25DH1, low activity with isoenzyme S25DH2. No activity with isoenzyme S25DH3 and S25DH4 | Sterolibacterium denitrificans | 25-hydroxy-vitamin D3 + reduced acceptor | - |
? |
Subunits | Comment | Organism |
---|---|---|
dodecamer | alpha4beta4gamma4, isoenzyme S25DH4 | Sterolibacterium denitrificans |
nonamer | alpha3beta3gamma3, isoenzyme S25DH3 | Sterolibacterium denitrificans |
octamer | alpha2beta3gamma3, isoenzyme S25DH2 | Sterolibacterium denitrificans |
Synonyms | Comment | Organism |
---|---|---|
S25DH | - |
Sterolibacterium denitrificans |
S25DH1 | - |
Sterolibacterium denitrificans |
S25DH2 | - |
Sterolibacterium denitrificans |
S25DH3 | - |
Sterolibacterium denitrificans |
S25DH4 | - |
Sterolibacterium denitrificans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
campest-4-en-3-one | isoenzyme S25DH3, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.14 | - |
beta-sitost-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.17 | - |
campest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.21 | - |
cholest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.23 | - |
vitamin D3 | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.29 | - |
cholest-4-en-3-one | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.33 | - |
7-dehydrocholesterol | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
1.11 | - |
cholest-4-en-3-one | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
heme b | the enzyme contains heme B | Sterolibacterium denitrificans | |
[3Fe-4S]-center | the enzyme contains one [3Fe-4S] cluster | Sterolibacterium denitrificans | |
[4Fe-4S]-center | the enzyme contains four [4Fe-4S] clusters | Sterolibacterium denitrificans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
campest-4-en-3-one | isoenzyme S25DH3, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.46 | - |
cholest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.5 | - |
campest-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
0.79 | - |
vitamin D3 | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans | |
1.2 | - |
beta-sitost-4-en-3-one | isoenzyme S25DH4, pH 7.0, 30°C | Sterolibacterium denitrificans | |
2.3 | - |
cholest-4-en-3-one | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
2.7 | - |
7-dehydrocholesterol | isoenzyme S25DH2, pH 7.0, 30°C | Sterolibacterium denitrificans | |
2.8 | - |
cholest-4-en-3-one | isoenzyme S25DH1, pH 7.0, 30°C | Sterolibacterium denitrificans |