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Literature summary for 1.14.15.4 extracted from

  • Zuidweg, M.H.J.
    Hydroxylation of Reichstein's compound S with cell-free preparations from Curvularia lunata (1968), Biochim. Biophys. Acta, 152, 144-158.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
KCN slight Curvularia lunata
MnCl2 causes most of activity to disappear in crude extracts Curvularia lunata
NAD(P)H enhance activity of crude extracts that show hydroxylation without any cofactor Curvularia lunata

General Stability

General Stability Organism
alpha-lipoic acid, during DEAE chromatography stabilizes Curvularia lunata
EDTA, stabilizes during purification Curvularia lunata
GSH, stabilizes during purification, as well as other sulfhydryl compounds Curvularia lunata
Tween 80, positive influence on activity conservation during extraction and fractionation Curvularia lunata

Inhibitors

Inhibitors Comment Organism Structure
Metyrapone 2 mM Curvularia lunata
p-chloromercuribenzoate 1 mM Curvularia lunata
phenazine methosulfate 1 mM, crude extract Curvularia lunata

Organism

Organism UniProt Comment Textmining
Curvularia lunata
-
Boedijn NRRL 2380
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, ion-exchange chromatography Curvularia lunata

Reaction

Reaction Comment Organism Reaction ID
a steroid + 2 reduced adrenodoxin + O2 + 2 H+ = an 11beta-hydroxysteroid + 2 oxidized adrenodoxin + H2O both hydroxylation activities seem to occur in a single enzyme Curvularia lunata

Storage Stability

Storage Stability Organism
-30°C, extract precipitate stable, solution less stable, DEAE fraction unstable Curvularia lunata
-30°C, preserved by Tween 80, conservation of activity Curvularia lunata
0°C, preserved by Tween 80, conservation of activity Curvularia lunata
2°C, crude extract loses 50% activity in 24 h Curvularia lunata
purified or partially purified enzyme unstable during storage Curvularia lunata

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
17alpha,21-dihydroxy-pregn-4-ene-3,20-dione + reduced adrenal ferredoxin + O2 i.e. Reichstein's compound, inducer Curvularia lunata 11beta,17alpha,21-trihydroxy-pregn-4-ene-3,20-dione + oxidized adrenal ferredoxin + H2O hydrocortisone, 60%, 11beta hydroxylation ?
17alpha,21-dihydroxy-pregn-4-ene-3,20-dione + reduced adrenal ferredoxin + O2 i.e. Reichstein's compound, inducer Curvularia lunata 14alpha,17alpha,21-trihydroxy-pregn-4-ene-3,20-dione + oxidized adrenal ferredoxin + H2O 25%, 14alpha hydroxylation ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
assay at Curvularia lunata

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2 15 2fold decrease of activity with increase of temperature, temperatures above 25°C unfavourable Curvularia lunata

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2
-
crude extract, 7 mg/ml, 24 h, loss of half the activity, DEAE fraction unstable Curvularia lunata

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Curvularia lunata

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Curvularia lunata