Literature summary 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.
Activating Compound
| EC Number |
Activating Compound |
Comment |
Organism |
Structure |
|---|
| 1.14.15.4 |
KCN |
slight |
Curvularia lunata |
|
| 1.14.15.4 |
MnCl2 |
causes most of activity to disappear in crude extracts |
Curvularia lunata |
|
| 1.14.15.4 |
NAD(P)H |
enhance activity of crude extracts that show hydroxylation without any cofactor |
Curvularia lunata |
|
General Stability
| EC Number |
General Stability |
Organism |
|---|
| 1.14.15.4 |
alpha-lipoic acid, during DEAE chromatography stabilizes |
Curvularia lunata |
| 1.14.15.4 |
EDTA, stabilizes during purification |
Curvularia lunata |
| 1.14.15.4 |
GSH, stabilizes during purification, as well as other sulfhydryl compounds |
Curvularia lunata |
| 1.14.15.4 |
Tween 80, positive influence on activity conservation during extraction and fractionation |
Curvularia lunata |
Inhibitors
| EC Number |
Inhibitors |
Comment |
Organism |
Structure |
|---|
| 1.14.15.4 |
Metyrapone |
2 mM |
Curvularia lunata |
|
| 1.14.15.4 |
p-chloromercuribenzoate |
1 mM |
Curvularia lunata |
|
| 1.14.15.4 |
phenazine methosulfate |
1 mM, crude extract |
Curvularia lunata |
|
Organism
| EC Number |
Organism |
UniProt |
Comment |
Textmining |
|---|
| 1.14.15.4 |
Curvularia lunata |
- |
Boedijn NRRL 2380 |
- |
Purification (Commentary)
| EC Number |
Purification (Comment) |
Organism |
|---|
| 1.14.15.4 |
ammonium sulfate fractionation, ion-exchange chromatography |
Curvularia lunata |
Reaction
| EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
|---|
| 1.14.15.4 |
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
| EC Number |
Storage Stability |
Organism |
|---|
| 1.14.15.4 |
-30°C, extract precipitate stable, solution less stable, DEAE fraction unstable |
Curvularia lunata |
| 1.14.15.4 |
-30°C, preserved by Tween 80, conservation of activity |
Curvularia lunata |
| 1.14.15.4 |
0°C, preserved by Tween 80, conservation of activity |
Curvularia lunata |
| 1.14.15.4 |
2°C, crude extract loses 50% activity in 24 h |
Curvularia lunata |
| 1.14.15.4 |
purified or partially purified enzyme unstable during storage |
Curvularia lunata |
Substrates and Products (Substrate)
| EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
|---|
| 1.14.15.4 |
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 |
? |
|
| 1.14.15.4 |
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]
| EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
|---|
| 1.14.15.4 |
20 |
- |
assay at |
Curvularia lunata |
Temperature Range [°C]
| EC Number |
Temperature Minimum [°C] |
Temperature Maximum [°C] |
Comment |
Organism |
|---|
| 1.14.15.4 |
2 |
15 |
2fold decrease of activity with increase of temperature, temperatures above 25°C unfavourable |
Curvularia lunata |
Temperature Stability [°C]
| EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
|---|
| 1.14.15.4 |
2 |
- |
crude extract, 7 mg/ml, 24 h, loss of half the activity, DEAE fraction unstable |
Curvularia lunata |
pH Optimum
| EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
|---|
| 1.14.15.4 |
8 |
- |
- |
Curvularia lunata |
Cofactor
| EC Number |
Cofactor |
Comment |
Organism |
Structure |
|---|
| 1.14.15.4 |
NADPH |
- |
Curvularia lunata |
|
