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Literature summary for 1.1.1.149 extracted from

  • Krafft, A.E.; Hylemon, P.B.
    Purification and characterization of a novel form of 20alpha-hydroxysteroid dehydrogenase from Clostridium scindens (1989), J. Bacteriol., 171, 2925-2932.
    View publication on PubMedView publication on EuropePMC
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Crystallization (Commentary)

Crystallization (Comment) Organism
-
 [Clostridium] scindens

General Stability

General Stability Organism
2-mercaptoethanol for stabilization during purification [Clostridium] scindens
glycerol for prevention of large activity loss [Clostridium] scindens

Inhibitors

Inhibitors Comment Organism Structure
NADH concentrations above 0.1 mM [Clostridium] scindens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0081
-
 NADH cortisone 0.05 mM [Clostridium] scindens
0.022
-
 cortisone NADH, pH 6.3 [Clostridium] scindens
0.032
-
 cortisol NADH, pH 6.3 [Clostridium] scindens
0.041
-
 20alpha-dihydrocortisol NAD+, pH 7.9 [Clostridium] scindens
0.526
-
 NAD+ 20alpha-dihydrocortisol [Clostridium] scindens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
 4 * 40000, identical, SDS-PAGE, N-terminal amino acid sequence [Clostridium] scindens
160000
-
 +/-2000 SDS-PAGE, PAGE, gel filtration-HPLC [Clostridium] scindens

Organism

Organism UniProt Comment Textmining
[Clostridium] scindens
-
 ATCC 35704
-

Purification (Commentary)

Purification (Comment) Organism
anion exchange and affinity chromatography, gel filtration, HPLC [Clostridium] scindens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.282
-
 measured with cortisone [Clostridium] scindens

Storage Stability

Storage Stability Organism
4°C or -20°C, above 0.5 mg of purified protein per ml, 50 mM phosphate buffer, pH 6.8, 10 mM 2-mercaptoethanol, 50% glycerol, v/v, several days [Clostridium] scindens
4°C pH 6-7 [Clostridium] scindens
no activity after single freeze-thaw of enzyme at any stage [Clostridium] scindens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
11-deoxycortisol + NADH
-
 [Clostridium] scindens ? + NAD+
-
 ?
17alpha,21-dihydroxyadrenocorticosteroid + NAD(P)H NADH preferred [Clostridium] scindens ? + NADP+
-
 ?
5beta-dihydrocortisol + NADH
-
 [Clostridium] scindens ? + NAD+
-
 r
cortisol + NADH
-
 [Clostridium] scindens 20alpha-dihydrocortisol + NAD+
-
 r
cortisone + NADH
-
 [Clostridium] scindens ? + NAD+
-
 ?

Subunits

Subunits Comment Organism
tetramer 4 * 40000, identical, SDS-PAGE, N-terminal amino acid sequence [Clostridium] scindens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 6.5 steroid reduction e.g. 20alpha-dihydrocortisol with MES, MOPS or sodium phosphate, NADH [Clostridium] scindens
8
-
 steroid oxidation e.g. cortisol with Tris hydrochloride, NAD+ [Clostridium] scindens

pH Range

pH Minimum pH Maximum Comment Organism
5.4 7.6 steroid reduction e.g. 20alpha-dihydrocortisol with MES, MOPS or sodium phosphate, NADH [Clostridium] scindens
6.3 9 steroid oxidation e.g. cortisol with Tris hydrochloride, NAD+ [Clostridium] scindens

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 [Clostridium] scindens
NADH preferred under optimum conditions for reduction of cortisol [Clostridium] scindens
NADPH 50% of NADH-activity [Clostridium] scindens