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Literature summary for 1.14.15.30 extracted from

  • Zhang, X.; Rao, Z.; Zhang, L.; Xu, M.; Yang, T.
    Efficient 9alpha-hydroxy-4-androstene-3,17-dione production by engineered Bacillus subtilis co-expressing Mycobacterium neoaurum 3-ketosteroid 9alpha-hydroxylase and B. subtilis glucose 1-dehydrogenase with NADH regeneration (2016), SpringerPlus, 5, 1207 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.15.30

Cloned(Commentary)

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

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

Inhibitors Comment Organism Structure
Co2+
-
 Mycolicibacterium neoaurum
Cu2+
-
 Mycolicibacterium neoaurum
Fe3+
-
 Mycolicibacterium neoaurum
Ni2+
-
 Mycolicibacterium neoaurum
Zn2+
-
 Mycolicibacterium neoaurum

Natural Substrates/ Products (Substrates)

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

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 (Commentary)

Purification (Comment) Organism
recombinant His-tagged KshA and KshB from Bacillus subtilis strain 168 by nickel affinity chromatography Mycolicibacterium neoaurum

Specific Activity [micromol/min/mg]

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

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 and Products (Substrate)

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

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
33
-
-
 Mycolicibacterium neoaurum
33
-
 recombinant enzyme Mycolicibacterium neoaurum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Mycolicibacterium neoaurum
7
-
 assay at Mycolicibacterium neoaurum

General Information

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