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Literature summary extracted from

  • Petrusma, M.; Dijkhuizen, L.; van Der Geize, R.
    Structural features in the KshA terminal oxygenase protein that determine substrate preference of 3-ketosteroid 9alpha-hydroxylase enzymes (2012), J. Bacteriol., 194, 115-121.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.30 expressed in Escherichia coli C41(DE3) cells Rhodococcus rhodochrous
1.14.15.30 expression of chimeric kshA mutant genes, KshA1A5beta and KshA5A1beta, in Escherichia coli, coexpression of the His-tagged KshA mutants with His-tagged KshB, molar ratio of KshA5A1/KshB is 1:8, low expression level of soluble KshA5A1alpha enzyme Rhodococcus rhodochrous

Protein Variants

EC Number Protein Variants Comment Organism
1.14.15.30 D230E/S232T/F238Y site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 D230E/S232T/F238Y the mutant of isoform KshA1 shows no 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 D242W site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 D242W the mutant of isoform KshA1 exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 E236D/T238S/Y244F site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 E236D/T238S/Y244F the mutant of isoform KshA5 shows no 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 additional information mutant KshA1A5beta is isoform KshA1 with beta sheet of KshA5 and the mutations kshA1 E198 to V255 exchanged for kshA5 E204 to I261, and exhibits 3-ketosteroid 9alpha-hydroxylase activity. Mutant KshA1A5alpha is isoform KshA1 with alpha helix of KshA5 and mutations kshA1 I257 to I306 exchanged for kshA5 I263 to V312, and shows not enzymatic activity. Mutant KshA1A5loop is isoform KshA1 with loop region of KshA5 and mutation kshA1 209-QAREDTRPHANGQPKMIGS-227 exchanged for kshA5 215-TGREDVISGTNYDDPNAEL-233, and exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 additional information mutant KshA5A1beta is isoform KshA5 with beta sheet of KshA1 and mutation kshA5 E204 to I261 exchanged for kshA1 E198 to V255, and exhibits 3-ketosteroid 9alpha-hydroxylase activity. Mutant KshA5A1alpha is isoform KshA5 with alpha helix of KshA1 and mutation kshA5 I263 to V312 exchanged for kshA1 I257 to I306, and shows initial 3-ketosteroid 9alpha-hydroxylase activity. Mutant KshA5A1loop is isoform KshA5 with loop region of KshA1 and mutation kshA5 215-TGREDVISGTNYDDPNAEL-233 exchanged for kshA1 209-QAREDTRPHANGQPKMIGS-227, and exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 additional information construction of chimeric KshA enzyme mutants with exchanged helix-grip fold beta-sheets that retain KSH activity. kshA1 E198 to V255 is exchanged for kshA5 E204 to I261. kshA1 I257 to I306 is exchanged for kshA5 I263 to V312, kshA5 E204 to I261 is exchanged for kshA1 E198 to V255, kshA5 I263 to V312 is exchanged for kshA1 I257 to I306, kshA1 209-QAREDTRPHANGQPKMIGS-227is exchanged for kshA5 215-TGREDVISGTNYDDPNAEL-233, and kshA5 215-TGREDVISGTNYDDPNAEL-233 is exchanged for kshA1 209-QAREDTRPHANGQPKMIGS-227 Rhodococcus rhodochrous
1.14.15.30 Q209T/A210G site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 Q209T/A210G the mutant of isoform KshA1 exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 T207V/T209S/Y211F/H213R/S214G/T215Q/G216A site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 T207V/T209S/Y211F/H213R/S214G/T215Q/G216A the mutant of isoform KshA5 exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 V201T/S203T/F205Y/R207H/G208S/Q209T/A210G site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 V201T/S203T/F205Y/R207H/G208S/Q209T/A210G the mutant of isoform KshA1 exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous
1.14.15.30 W248D site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme Rhodococcus rhodochrous
1.14.15.30 W248D the mutant of isoform KshA5 exhibits 3-ketosteroid 9alpha-hydroxylase activity Rhodococcus rhodochrous

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.15.30 additional information
-
additional information kinetic analysis of the activity of KshA mutants with 4-23,24-bisnorcholesta-4-ene-22-oic acid Rhodococcus rhodochrous
1.14.15.30 0.01
-
23,24-bisnorcholesta-4-ene-22-oic acid Km less than 0.01 mM, isoform KshA1, in 50mMTris-HCl buffer (pH 7.0), temperature not specified in the publication Rhodococcus rhodochrous
1.14.15.30 0.01
-
23,24-bisnorcholesta-4-ene-22-oic acid Km less than 0.01 mM, isoform KshA5, in 50 mM Tris-HCl buffer (pH 7.0), temperature not specified in the publication Rhodococcus rhodochrous

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.15.30 Fe2+ the enzyme contains a nonheme Fe2+ Rhodococcus rhodochrous

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.15.30 androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 Rhodococcus rhodochrous
-
9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.30 Rhodococcus rhodochrous F1CMX0 gene kshA1, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase
-
1.14.15.30 Rhodococcus rhodochrous F1CMX0 isozyme KshA1
-
1.14.15.30 Rhodococcus rhodochrous F1CMX3 gene kshB, encoding the ferredoxin reductase
-
1.14.15.30 Rhodococcus rhodochrous F1CMY8 gene kshA5, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase
-
1.14.15.30 Rhodococcus rhodochrous F1CMY8 isozyme KshA5
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMX0 gene kshA1, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMX0 isozyme KshA1
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMX3 gene kshB, encoding the ferredoxin reductase
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMY8 gene kshA5, encoding an isoform of the oxygenase component of 3-ketosteroid 9alpha-hydroxylase
-
1.14.15.30 Rhodococcus rhodochrous DSM 43269 F1CMY8 isozyme KshA5
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.15.30 copurification of recombinant His-tagged KshA mutants with His-tagged KshB Rhodococcus rhodochrous

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.30 1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 244% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous ? + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 51% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous ? + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 244% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous DSM 43269 ? + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1,4-androstadiene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 51% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous DSM 43269 ? + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 23% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 99% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 1-(5alpha)-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous DSM 43269 9alpha-hydroxy-5alpha-androst-1-en-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 11beta-hydrocortisone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-11beta-hydrocortisone + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 11beta-hydrocortisone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 93% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-11beta-hydrocortisone + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 11beta-hydrocortisone + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous DSM 43269 9alpha-hydroxy-11beta-hydrocortisone + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. nordion, the wild type homologue KshA1 shows 24% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. nordion, the wild type homologue KshA5 shows 111% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 19-nor-4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous DSM 43269 9alpha-hydroxy-19-nor-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 548% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 88% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous DSM 43269 9alpha-hydroxy-3-oxo-23,24-bisnorcholest-4-en-22-oic acid + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. testosterone, the wild type homologue KshA1 shows 158% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. testosterone, the wild type homologue KshA5 shows 113% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous DSM 43269 9alpha,17beta-dihydroxy-4-androstene-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 100% activity Rhodococcus rhodochrous 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-androstene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 100% activity Rhodococcus rhodochrous 9alpha-hydroxy-4-androstene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. progesterone, the wild type homologue KshA1 shows 372% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 4-pregnene-3,20-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. progesterone, the wild type homologue KshA5 shows 65% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-4-pregnene-3,20-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5alpha-androstane-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha,17beta-dihydroxy-5alpha-androstane-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5alpha-androstane-17beta-ol-3-one + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 i.e. stanolon, the wild type homologue KshA5 shows 100% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha,17beta-dihydroxy-5alpha-androstane-3-one + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 12% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5alpha-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 73% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5alpha-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA1 shows 19% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 5beta-androstane-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 the wild type homologue KshA5 shows 99% activity compared to 4-androstene-3,17-dione Rhodococcus rhodochrous 9alpha-hydroxy-5beta-androstane-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 androsta-1,4-diene-3,17-dione + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2
-
Rhodococcus rhodochrous 9alpha-hydroxyandrosta-1,4-diene-3,17-dione + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
?
1.14.15.30 additional information the KshA1 homologue has a narrow substrate range with a high preference for 23,24-bisnorcholesta-4-ene-22-oic acid and 23,24-bisnorcholesta-1,4-diene-22-oic acid and preference to a lesser extent for 1,4-androstadiene-3,17-dione and 4-pregnene-3,20-dione (progesterone) Rhodococcus rhodochrous ?
-
?
1.14.15.30 additional information the KshA5 homologue has a broad substrate range with no apparent preference for any of the tested steroids Rhodococcus rhodochrous ?
-
?
1.14.15.30 additional information the wild type homologue KshA1 shows no activity with 5alpha-androstane-17beta-ol-3-one (stanolon), 5-cholestene-3beta-ol (cholesterol), 3alpha-hydroxy-5alpha-pregnane-20-one, 11beta-hydrocortisone, 3beta-hydroxy-5alpha-androstane-17-one, and 9alpha-hydroxy-4-androstene-3,17-dione Rhodococcus rhodochrous ?
-
?
1.14.15.30 additional information the wild type homologue KshA5 shows no activity with 5-cholestene-3beta-ol, 3alpha-hydroxy-5beta-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, and 9alpha-hydroxy-4-androstene-3,17-dione Rhodococcus rhodochrous ?
-
?
1.14.15.30 additional information substrate specificities of KshA variants, overview. No activity by all enzymes with 3alpha-hydroxy-5alpha-pregnane-20-one, 9alpha-hydroxy-4-androstene-3,17-dione, 5-cholestene-3beta-ol, and 3beta-hydroxy-5alpha-androstane-17-one Rhodococcus rhodochrous ?
-
?
1.14.15.30 additional information the KshA1 homologue has a narrow substrate range with a high preference for 23,24-bisnorcholesta-4-ene-22-oic acid and 23,24-bisnorcholesta-1,4-diene-22-oic acid and preference to a lesser extent for 1,4-androstadiene-3,17-dione and 4-pregnene-3,20-dione (progesterone) Rhodococcus rhodochrous DSM 43269 ?
-
?
1.14.15.30 additional information the wild type homologue KshA1 shows no activity with 5alpha-androstane-17beta-ol-3-one (stanolon), 5-cholestene-3beta-ol (cholesterol), 3alpha-hydroxy-5alpha-pregnane-20-one, 11beta-hydrocortisone, 3beta-hydroxy-5alpha-androstane-17-one, and 9alpha-hydroxy-4-androstene-3,17-dione Rhodococcus rhodochrous DSM 43269 ?
-
?
1.14.15.30 additional information substrate specificities of KshA variants, overview. No activity by all enzymes with 3alpha-hydroxy-5alpha-pregnane-20-one, 9alpha-hydroxy-4-androstene-3,17-dione, 5-cholestene-3beta-ol, and 3beta-hydroxy-5alpha-androstane-17-one Rhodococcus rhodochrous DSM 43269 ?
-
?
1.14.15.30 additional information the KshA5 homologue has a broad substrate range with no apparent preference for any of the tested steroids Rhodococcus rhodochrous DSM 43269 ?
-
?
1.14.15.30 additional information the wild type homologue KshA5 shows no activity with 5-cholestene-3beta-ol, 3alpha-hydroxy-5beta-pregnane-20-one, 3beta-hydroxy-5alpha-androstane-17-one, and 9alpha-hydroxy-4-androstene-3,17-dione Rhodococcus rhodochrous DSM 43269 ?
-
?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.30 3-ketosteroid 9alpha-hydroxylase
-
Rhodococcus rhodochrous
1.14.15.30 KSH
-
Rhodococcus rhodochrous
1.14.15.30 KshA terminal oxygenase component of 3-ketosteroid 9alpha-hydroxylase Rhodococcus rhodochrous
1.14.15.30 KshA1 isoform Rhodococcus rhodochrous
1.14.15.30 KshA5 isoform Rhodococcus rhodochrous
1.14.15.30 KshAB
-
Rhodococcus rhodochrous
1.14.15.30 KshB reductase component of 3-ketosteroid 9alpha-hydroxylase Rhodococcus rhodochrous

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.15.30 7
-
assay at Rhodococcus rhodochrous

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.30 additional information a Rieske nonheme monooxygenase Rhodococcus rhodochrous
1.14.15.30 [2Fe-2S]-center
-
Rhodococcus rhodochrous

General Information

EC Number General Information Comment Organism
1.14.15.30 additional information determination of the enzyme's substrate preference, three-dimensional structure modeling of KshA1, overview. The variable loop within the beta-sheet plays a role in the substrate preference of KshA enzymes Rhodococcus rhodochrous
1.14.15.30 additional information determination of the enzyme's substrate preference, three-dimensional structure modeling of KshA5, overview. The variable loop within the beta-sheet plays a role in the substrate preference of KshA enzymes Rhodococcus rhodochrous