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Literature summary for 1.17.99.10 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)

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

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 [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

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

Molecular Weight [Da]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Sterolibacterium denitrificans
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Sterolibacterium denitrificans

Substrates and Products (Substrate)

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

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

Synonyms

Synonyms Comment Organism
S25DH
-
Sterolibacterium denitrificans
S25DH1
-
Sterolibacterium denitrificans
S25DH2
-
Sterolibacterium denitrificans
S25DH3
-
Sterolibacterium denitrificans
S25DH4
-
Sterolibacterium denitrificans

Turnover Number [1/s]

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

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 [mM/s]

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