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Literature summary extracted from

  • Inazu, A.; Sato, K.; Nakayama, T.; Deyashiki, Y.; Hara, A.; Nozawa, Y.
    Purification and characterization of a novel dimeric 20alpha-hydroxysteroid dehydrogenase from Tetrahymena pyriformis (1994), Biochem. J., 297, 195-200.
    View publication on PubMedView publication on EuropePMC

General Stability

EC Number General Stability Organism
1.1.1.149 20%, v/v, glycerol for stabilization during purification Tetrahymena pyriformis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.149 barbiturates
-
 Tetrahymena pyriformis
1.1.1.149 Cu2+ 40% inhibition Tetrahymena pyriformis
1.1.1.149 Diphenic acid less than 17% inhibition Tetrahymena pyriformis
1.1.1.149 indomethacin less than 17% inhibition Tetrahymena pyriformis
1.1.1.149 additional information no inhibition with thiol reagents, reducing or chelating agents, ADH inhibitors Tetrahymena pyriformis
1.1.1.149 natural oestrogens e.g. oestrone less than 15% inhibition Tetrahymena pyriformis
1.1.1.149 quercitrin
-
 Tetrahymena pyriformis
1.1.1.149 synthetic oestrogens e.g. hexestrol, silbestrol, dienstrol Tetrahymena pyriformis
1.1.1.149 valporic acid less than 17% inhibition Tetrahymena pyriformis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.149 0.0002
-
 NADPH 20-oxo and 20alpha-hydroxy-steroids, pH 7.0 Tetrahymena pyriformis
1.1.1.149 0.0029
-
 17alpha-hydroxyprogesterone
-
 Tetrahymena pyriformis
1.1.1.149 0.0034
-
 17alpha-hydroxypregnenolone
-
 Tetrahymena pyriformis
1.1.1.149 0.008
-
 NADP+ 20-oxo and 20alpha-hydroxy-steroids, pH 7.0 Tetrahymena pyriformis
1.1.1.149 0.013
-
 17alpha,20alpha-dihydroxypregn-4-en-3-one
-
 Tetrahymena pyriformis
1.1.1.149 0.02
-
 17alpha,21-dihydroxy-5beta-pregnane-3,11,20-trione
-
 Tetrahymena pyriformis
1.1.1.149 0.022
-
 17alpha,21-dihydroxy-5beta-pregnan-3,20-dione
-
 Tetrahymena pyriformis
1.1.1.149 0.064
-
 isatin
-
 Tetrahymena pyriformis
1.1.1.149 0.067
-
 ethyl acetoacetate
-
 Tetrahymena pyriformis
1.1.1.149 0.073
-
 20alpha-hydroxy-5beta-pregnan-3-one
-
 Tetrahymena pyriformis
1.1.1.149 0.078
-
 Camphorquinone
-
 Tetrahymena pyriformis
1.1.1.149 0.081
-
 Pyridine-3-aldehyde
-
 Tetrahymena pyriformis
1.1.1.149 0.125
-
 1,2-Cyclohexanedione
-
 Tetrahymena pyriformis
1.1.1.149 0.21
-
 2,4-Pentanedione
-
 Tetrahymena pyriformis
1.1.1.149 0.34
-
 4-nitrobenzaldehyde
-
 Tetrahymena pyriformis
1.1.1.149 0.43
-
 Phenylglyoxal
-
 Tetrahymena pyriformis
1.1.1.149 0.67
-
 D-glyceraldehyde
-
 Tetrahymena pyriformis
1.1.1.149 2.1
-
 benzene dihydrodiol
-
 Tetrahymena pyriformis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.149 33000
-
 2 * 33000, SDS-PAGE Tetrahymena pyriformis
1.1.1.149 68000
-
 gel filtration Tetrahymena pyriformis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.149 additional information Tetrahymena pyriformis role of enzyme suggested to be for detoxication or metabolism of carbonyl compounds because of broad substrate specificity ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.149 Tetrahymena pyriformis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.149 ammonium sulfate precipitation, ultrafiltration, chromatography Tetrahymena pyriformis

Reaction

EC Number Reaction Comment Organism Reaction ID
1.1.1.149 17alpha,20alpha-dihydroxypregn-4-en-3-one + NAD(P)+ = 17alpha-hydroxyprogesterone + NAD(P)H + H+ aldoketo reductase superfamily Tetrahymena pyriformis
a

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 6.2 6.9
-
 Tetrahymena pyriformis

Storage Stability

EC Number Storage Stability Organism
1.1.1.149 -20°C, purified enzyme, 10 mM Tris/HCl, pH 7.5, 0.5 mM EDTA, 1 mM 2-mercaptoethanol, 20%, v/v, glycerol, 2 weeks, little activity loss Tetrahymena pyriformis
1.1.1.149 4°C, Sephadex G-100 fraction, rapid decrease, 2 days, 50% activity loss, 80% activity remaining by addition of 20%, v/v, glycerol Tetrahymena pyriformis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.149 1,2-cyclohexanedione + NADPH + H+
-
 Tetrahymena pyriformis ?
-
 r
1.1.1.149 17alpha,21-dihydroxy-5beta-pregnan-3,20-dione + NADPH + H+
-
 Tetrahymena pyriformis 17alpha,20alpha,21-trihydroxy-5beta-pregnane-3-one + NADP+
-
 r
1.1.1.149 17alpha,21-dihydroxy-5beta-pregnane-3,11,20-trione + NADPH + H+
-
 Tetrahymena pyriformis 17alpha,20alpha,21-trihydroxy-5beta-pregnane-3,11-dione + NADP+
-
 r
1.1.1.149 17alpha-hydroxypregnenolone + NADPH
-
 Tetrahymena pyriformis ? + NADP+
-
 ?
1.1.1.149 17alpha-hydroxyprogesterone + NADPH + H+ best substrate Tetrahymena pyriformis 17alpha,20alpha-dihydroxy-pregn-4-en-3-one + NADP+
-
 r
1.1.1.149 2,4-pentanedione + NADPH + H+
-
 Tetrahymena pyriformis 4-hydroxypentan-2-one + NADP+
-
 r
1.1.1.149 20alpha-hydroxy-5beta-pregnan-3-one + NADP+
-
 Tetrahymena pyriformis 5beta-pregnane-3,20-dione + NADPH
-
 ?
1.1.1.149 4-nitrobenzaldehyde + NADPH
-
 Tetrahymena pyriformis 4-nitrobenzyl alcohol + NADP+
-
 ?
1.1.1.149 benzene dihydrodiol + NADP+
-
 Tetrahymena pyriformis 2-hydroxycyclohexa-3,5-diene-1-one + NADPH + H+
-
 r
1.1.1.149 camphorquinone + NADP+
-
 Tetrahymena pyriformis ?
-
 r
1.1.1.149 D-glyceraldehyde + NADPH + H+
-
 Tetrahymena pyriformis glycerol + NADP+
-
 r
1.1.1.149 ethyl acetoaetate + NADPH
-
 Tetrahymena pyriformis (S)-ethyl 3-hydroxybutyrate + NADP+
-
 ?
1.1.1.149 isatin + NADPH
-
 Tetrahymena pyriformis NADP+ + ?
-
 ?
1.1.1.149 additional information less or no activity with 5beta-androstane-3,17-dione, 17beta-hydroxy-5beta-androstan-3-one, progesterone, 5beta-pregnane-3,20-dione, cortisone oestrone, 20-oxopregnenes without 17alpha-hydroxy group, 20-oxopregnanes, corticosteroides, 3- or 17-oxosteroids, 20beta-hydroxysteroids, 3- or 17-hydroxysteroids, alcohols as substrate, other substrates: isatin, ethyl acetoacetate, camphorquinone, pyridine-3-aldehyde, 1,2-cyclohexanedione, 2,4-pentanedione, 4-nitrobenzaldehyde, benzene-dihydrodiol Tetrahymena pyriformis ?
-
 ?
1.1.1.149 additional information role of enzyme suggested to be for detoxication or metabolism of carbonyl compounds because of broad substrate specificity Tetrahymena pyriformis ?
-
 ?
1.1.1.149 phenylglyoxal + NADPH + H+
-
 Tetrahymena pyriformis ?
-
 r
1.1.1.149 pyridine-3-aldehyde + NADPH
-
 Tetrahymena pyriformis NADP+ + ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.149 dimer 2 * 33000, SDS-PAGE Tetrahymena pyriformis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.149 25
-
 10 min at pH 6-10 Tetrahymena pyriformis
1.1.1.149 40
-
 up to, for 5 min 95% initial activity, rapidly inactivated above 45°C Tetrahymena pyriformis
1.1.1.149 45
-
 rapidly inactivated above Tetrahymena pyriformis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.149 6 6.5 17alpha-hydroxyprogesterone reduction Tetrahymena pyriformis
1.1.1.149 10.5
-
 steroid oxidation Tetrahymena pyriformis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.149 5.8 10.5 25°C, below and above range inactivation Tetrahymena pyriformis

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.149 4.5 11.5 25°C, below and above range total loss of activity Tetrahymena pyriformis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.149 NADP+ strictly specific Tetrahymena pyriformis
1.1.1.149 NADPH strictly specific Tetrahymena pyriformis