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Literature summary for 1.1.1.149 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

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

Inhibitors

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

KM Value [mM]

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Tetrahymena pyriformis
-
-
-

Purification (Commentary)

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

Reaction

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.2 6.9
-
Tetrahymena pyriformis

Storage Stability

Storage Stability Organism
-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
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,2-cyclohexanedione + NADPH + H+
-
Tetrahymena pyriformis ?
-
r
17alpha,21-dihydroxy-5beta-pregnan-3,20-dione + NADPH + H+
-
Tetrahymena pyriformis 17alpha,20alpha,21-trihydroxy-5beta-pregnane-3-one + NADP+
-
r
17alpha,21-dihydroxy-5beta-pregnane-3,11,20-trione + NADPH + H+
-
Tetrahymena pyriformis 17alpha,20alpha,21-trihydroxy-5beta-pregnane-3,11-dione + NADP+
-
r
17alpha-hydroxypregnenolone + NADPH
-
Tetrahymena pyriformis ? + NADP+
-
?
17alpha-hydroxyprogesterone + NADPH + H+ best substrate Tetrahymena pyriformis 17alpha,20alpha-dihydroxy-pregn-4-en-3-one + NADP+
-
r
2,4-pentanedione + NADPH + H+
-
Tetrahymena pyriformis 4-hydroxypentan-2-one + NADP+
-
r
20alpha-hydroxy-5beta-pregnan-3-one + NADP+
-
Tetrahymena pyriformis 5beta-pregnane-3,20-dione + NADPH
-
?
4-nitrobenzaldehyde + NADPH
-
Tetrahymena pyriformis 4-nitrobenzyl alcohol + NADP+
-
?
benzene dihydrodiol + NADP+
-
Tetrahymena pyriformis 2-hydroxycyclohexa-3,5-diene-1-one + NADPH + H+
-
r
camphorquinone + NADP+
-
Tetrahymena pyriformis ?
-
r
D-glyceraldehyde + NADPH + H+
-
Tetrahymena pyriformis glycerol + NADP+
-
r
ethyl acetoaetate + NADPH
-
Tetrahymena pyriformis (S)-ethyl 3-hydroxybutyrate + NADP+
-
?
isatin + NADPH
-
Tetrahymena pyriformis NADP+ + ?
-
?
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 ?
-
?
additional information role of enzyme suggested to be for detoxication or metabolism of carbonyl compounds because of broad substrate specificity Tetrahymena pyriformis ?
-
?
phenylglyoxal + NADPH + H+
-
Tetrahymena pyriformis ?
-
r
pyridine-3-aldehyde + NADPH
-
Tetrahymena pyriformis NADP+ + ?
-
?

Subunits

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

Temperature Stability [°C]

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

pH Optimum

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

pH Range

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

pH Stability

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

Cofactor

Cofactor Comment Organism Structure
NADP+ strictly specific Tetrahymena pyriformis
NADPH strictly specific Tetrahymena pyriformis