Cloned (Comment) | Organism |
---|---|
expressed in Bacillus subtilis strain 168 | Mycolicibacterium neoaurum |
genes kshA and kshB, DNA and amino acid sequence determination and analysis, cloning into plasmid pMA5, recombinant expression of C-terminally His-tagged KshA and KshB in Bacillus subtilis strain 168, coexpression with Bacillus subtilis glucose 1-dehydrogenase, method development and evaluation, overview. KSH activity can only be detected when KshA and KshB are coexpressed | Mycolicibacterium neoaurum |
Protein Variants | Comment | Organism |
---|---|---|
additional information | efficient 9alpha-hydroxy-4-androstene-3,17-dione production by engineered Bacillus subtilis co-expressing Mycobacterium neoaurum 3-ketosteroid 9alpha-hydroxylase and Bacillus subtilis glucose 1-dehydrogenase with NADH regeneration | Mycolicibacterium neoaurum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Co2+ | - |
Mycolicibacterium neoaurum | |
Cu2+ | - |
Mycolicibacterium neoaurum | |
Fe3+ | - |
Mycolicibacterium neoaurum | |
Ni2+ | - |
Mycolicibacterium neoaurum | |
Zn2+ | - |
Mycolicibacterium neoaurum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycolicibacterium neoaurum | - |
9alpha-hydroxy-1,4-androstadiene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycolicibacterium neoaurum JC-12 | - |
9alpha-hydroxy-1,4-androstadiene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycolicibacterium neoaurum | - |
9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | Mycolicibacterium neoaurum JC-12 | - |
9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | Mycolicibacterium neoaurum | - |
9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | Mycolicibacterium neoaurum JC-12 | - |
9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
additional information | Mycolicibacterium neoaurum | Mycobacterium neoaurum strain JC-12 is a 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione producing strain | ? | - |
? | |
additional information | Mycolicibacterium neoaurum JC-12 | Mycobacterium neoaurum strain JC-12 is a 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione producing strain | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycolicibacterium neoaurum | A0A0F7JI94 AND D2Y640 | genes kshA and kshB encoding the enzyme subunits KshA and KshB | - |
Mycolicibacterium neoaurum | A0A0F7JI94 AND D2Y640 | subunit KshA and subunit KshB | - |
Mycolicibacterium neoaurum JC-12 | A0A0F7JI94 AND D2Y640 | genes kshA and kshB encoding the enzyme subunits KshA and KshB | - |
Mycolicibacterium neoaurum JC-12 | A0A0F7JI94 AND D2Y640 | subunit KshA and subunit KshB | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged KshA and KshB from Bacillus subtilis strain 168 by nickel affinity chromatography | Mycolicibacterium neoaurum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.57 | - |
recombinant enzyme from Bacillus subtilis, pH 7.0, 22°C | Mycolicibacterium neoaurum |
Storage Stability | Organism |
---|---|
-20°C and 0°C, 50 mM Tris-HCl buffer (pH 7.0), 24 h, 28% and 67% loss of activity, respectively | Mycolicibacterium neoaurum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycolicibacterium neoaurum | 9alpha-hydroxy-1,4-androstadiene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycolicibacterium neoaurum JC-12 | 9alpha-hydroxy-1,4-androstadiene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycolicibacterium neoaurum | 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Mycolicibacterium neoaurum JC-12 | 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Mycolicibacterium neoaurum | 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + H+ + O2 | - |
Mycolicibacterium neoaurum JC-12 | 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
additional information | Mycobacterium neoaurum strain JC-12 is a 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione producing strain | Mycolicibacterium neoaurum | ? | - |
? | |
additional information | KshB, the reductase component of KSH, accepts electrons from NADH and transfers it to oxygenase component KshA, the cooperation of KshA and KshB is critically important for maintaining KSH activity | Mycolicibacterium neoaurum | ? | - |
? | |
additional information | Mycobacterium neoaurum strain JC-12 is a 4-androstene-3,17-dione and 1,4-androstadiene-3,17-dione producing strain | Mycolicibacterium neoaurum JC-12 | ? | - |
? | |
additional information | KshB, the reductase component of KSH, accepts electrons from NADH and transfers it to oxygenase component KshA, the cooperation of KshA and KshB is critically important for maintaining KSH activity | Mycolicibacterium neoaurum JC-12 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 378000, subunit KshB, SDS-PAGE | Mycolicibacterium neoaurum |
? | x * 451000, subunit KshA, SDS-PAGE | Mycolicibacterium neoaurum |
More | enzyme KSH consists of KshA and KshB components | Mycolicibacterium neoaurum |
Synonyms | Comment | Organism |
---|---|---|
3-ketosteroid 9alpha-hydroxylase | - |
Mycolicibacterium neoaurum |
KSH | - |
Mycolicibacterium neoaurum |
KshA | reductase subunit | Mycolicibacterium neoaurum |
KshB | oxygenase subunit | Mycolicibacterium neoaurum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
33 | - |
- |
Mycolicibacterium neoaurum |
33 | - |
recombinant enzyme | Mycolicibacterium neoaurum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Mycolicibacterium neoaurum |
7 | - |
assay at | Mycolicibacterium neoaurum |
General Information | Comment | Organism |
---|---|---|
metabolism | 3-ketosteroid 9alpha-hydroxylase is a key enzyme in steroid metabolism and can catalyze the transformation of 4-androstene-3,17-dione to 9alpha-hydroxy-4-androstene-3,17-dione | Mycolicibacterium neoaurum |