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Literature summary 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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.149
-
 [Clostridium] scindens

General Stability

EC Number General Stability Organism
1.1.1.149 2-mercaptoethanol for stabilization during purification [Clostridium] scindens
1.1.1.149 glycerol for prevention of large activity loss [Clostridium] scindens

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.149 0.0081
-
 NADH cortisone 0.05 mM [Clostridium] scindens
1.1.1.149 0.022
-
 cortisone NADH, pH 6.3 [Clostridium] scindens
1.1.1.149 0.032
-
 cortisol NADH, pH 6.3 [Clostridium] scindens
1.1.1.149 0.041
-
 20alpha-dihydrocortisol NAD+, pH 7.9 [Clostridium] scindens
1.1.1.149 0.526
-
 NAD+ 20alpha-dihydrocortisol [Clostridium] scindens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.149 40000
-
 4 * 40000, identical, SDS-PAGE, N-terminal amino acid sequence [Clostridium] scindens
1.1.1.149 160000
-
 +/-2000 SDS-PAGE, PAGE, gel filtration-HPLC [Clostridium] scindens

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.149 [Clostridium] scindens
-
 ATCC 35704
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.149 0.282
-
 measured with cortisone [Clostridium] scindens

Storage Stability

EC Number Storage Stability Organism
1.1.1.149 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
1.1.1.149 4°C pH 6-7 [Clostridium] scindens
1.1.1.149 no activity after single freeze-thaw of enzyme at any stage [Clostridium] scindens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.149 11-deoxycortisol + NADH
-
 [Clostridium] scindens ? + NAD+
-
 ?
1.1.1.149 17alpha,21-dihydroxyadrenocorticosteroid + NAD(P)H NADH preferred [Clostridium] scindens ? + NADP+
-
 ?
1.1.1.149 5beta-dihydrocortisol + NADH
-
 [Clostridium] scindens ? + NAD+
-
 r
1.1.1.149 cortisol + NADH
-
 [Clostridium] scindens 20alpha-dihydrocortisol + NAD+
-
 r
1.1.1.149 cortisone + NADH
-
 [Clostridium] scindens ? + NAD+
-
 ?

Subunits

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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.149 6 6.5 steroid reduction e.g. 20alpha-dihydrocortisol with MES, MOPS or sodium phosphate, NADH [Clostridium] scindens
1.1.1.149 8
-
 steroid oxidation e.g. cortisol with Tris hydrochloride, NAD+ [Clostridium] scindens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.149 5.4 7.6 steroid reduction e.g. 20alpha-dihydrocortisol with MES, MOPS or sodium phosphate, NADH [Clostridium] scindens
1.1.1.149 6.3 9 steroid oxidation e.g. cortisol with Tris hydrochloride, NAD+ [Clostridium] scindens

Cofactor

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