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Literature summary extracted from

  • Jacoby, C.; Eipper, J.; Warnke, M.; Tiedt, O.; Mergelsberg, M.; Staerk, H.J.; Daus, B.; Martin-Moldes, Z.; Zamarro, M.T.; Diaz, E.; Boll, M.
    Four molybdenum-dependent steroid C-25 hydroxylases heterologous overproduction, role in steroid degradation, and application for 25-hydroxyvitamin D3 synthesis (2018), mBio, 9, e00694-18 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.17.99.10 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

EC Number General Stability Organism
1.17.99.10 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.17.99.10 0.12
-
 beta-sitost-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.123
-
 7-dehydrocholesterol isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.124
-
 cholest-4-en-3-one isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.29
-
 vitamin D3 isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.34
-
 campest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.39
-
 cholest-4-en-3-one isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.45
-
 cholest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 1.84
-
 campest-4-en-3-one isoenzyme S25DH3, pH 7.0, 30°C Sterolibacterium denitrificans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.17.99.10 Mo2+ the enzyme contains molybdenum Sterolibacterium denitrificans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.17.99.10 160000
-
 gel filtration, isoenzyme S25DH4 Sterolibacterium denitrificans
1.17.99.10 167000
-
 gel filtration, isoenzyme S25DH, isoenzyme S25DH2, isoenzyme S25DH4 Sterolibacterium denitrificans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.17.99.10 cholest-4-en-3-one + acceptor + H2O Sterolibacterium denitrificans
-
 25-hydroxycholest-4-en-3-one + reduced acceptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.17.99.10 Sterolibacterium denitrificans
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.17.99.10
-
 Sterolibacterium denitrificans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.17.99.10 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
-
 ?
1.17.99.10 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
-
 ?
1.17.99.10 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
-
 ?
1.17.99.10 cholest-4-en-3-one + acceptor + H2O
-
 Sterolibacterium denitrificans 25-hydroxycholest-4-en-3-one + reduced acceptor
-
 ?
1.17.99.10 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
-
 ?
1.17.99.10 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

EC Number Subunits Comment Organism
1.17.99.10 dodecamer alpha4beta4gamma4, isoenzyme S25DH4 Sterolibacterium denitrificans
1.17.99.10 nonamer alpha3beta3gamma3, isoenzyme S25DH3 Sterolibacterium denitrificans
1.17.99.10 octamer alpha2beta3gamma3, isoenzyme S25DH2 Sterolibacterium denitrificans

Synonyms

EC Number Synonyms Comment Organism
1.17.99.10 S25DH
-
 Sterolibacterium denitrificans
1.17.99.10 S25DH1
-
 Sterolibacterium denitrificans
1.17.99.10 S25DH2
-
 Sterolibacterium denitrificans
1.17.99.10 S25DH3
-
 Sterolibacterium denitrificans
1.17.99.10 S25DH4
-
 Sterolibacterium denitrificans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.17.99.10 0.018
-
 campest-4-en-3-one isoenzyme S25DH3, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.14
-
 beta-sitost-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.17
-
 campest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.21
-
 cholest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.23
-
 vitamin D3 isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.29
-
 cholest-4-en-3-one isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.33
-
 7-dehydrocholesterol isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 1.11
-
 cholest-4-en-3-one isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans

Cofactor

EC Number Cofactor Comment Organism Structure
1.17.99.10 heme b the enzyme contains heme B Sterolibacterium denitrificans
1.17.99.10 [3Fe-4S]-center the enzyme contains one [3Fe-4S] cluster Sterolibacterium denitrificans
1.17.99.10 [4Fe-4S]-center the enzyme contains four [4Fe-4S] clusters Sterolibacterium denitrificans

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.17.99.10 0.01
-
 campest-4-en-3-one isoenzyme S25DH3, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.46
-
 cholest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.5
-
 campest-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 0.79
-
 vitamin D3 isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 1.2
-
 beta-sitost-4-en-3-one isoenzyme S25DH4, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 2.3
-
 cholest-4-en-3-one isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 2.7
-
 7-dehydrocholesterol isoenzyme S25DH2, pH 7.0, 30°C Sterolibacterium denitrificans
1.17.99.10 2.8
-
 cholest-4-en-3-one isoenzyme S25DH1, pH 7.0, 30°C Sterolibacterium denitrificans