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Literature summary extracted from

  • Zhang, R.; Xu, X.; Cao, H.; Yuan, C.; Yuminaga, Y.; Zhao, S.; Shi, J.; Zhang, B.
    Purification, characterization, and application of a high activity 3-ketosteroid-DELTA1-dehydrogenase from Mycobacterium neoaurum DSM 1381 (2019), Appl. Microbiol. Biotechnol., 103, 6605-6616 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.99.4 gene KSTD2, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally or C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Mycolicibacterium neoaurum

General Stability

EC Number General Stability Organism
1.3.99.4 both the activity and stability of the enzyme are sensitive to various metal ions Mycolicibacterium neoaurum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.99.4 Ca2+ almost complete inhibition at 1 mM Mycolicibacterium neoaurum
1.3.99.4 Cu2+ almost complete inhibition at 1 mM Mycolicibacterium neoaurum
1.3.99.4 Fe3+ 40.6% inhibition at 1 mM Mycolicibacterium neoaurum
1.3.99.4 Mn2+ almost complete inhibition at 1 mM Mycolicibacterium neoaurum
1.3.99.4 additional information not affected by Na+, K+, and EDTA at 1 mM Mycolicibacterium neoaurum
1.3.99.4 Ni2+ 59.1% inhibition at 1 mM Mycolicibacterium neoaurum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.99.4 additional information
-
 additional information Michaelis-Menten kinetics Mycolicibacterium neoaurum
1.3.99.4 0.0158
-
 17alpha,21-dihydroxyprogesterone 21-acetate recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.0211
-
 17alpha-hydroxypregn-4-ene-3,20-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.0305
-
 16,17alpha-epoxyprogesterone recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.0687
-
 canrenone recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.1434
-
 4-androstene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.1733
-
 androsta-4,9(11)-diene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.1919
-
 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione acetate recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.8327
-
 9alpha-hydroxy-4-androstene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 0.9158
-
 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.3.99.4 Mg2+ slightly activating at 1 mM Mycolicibacterium neoaurum
1.3.99.4 additional information both the activity and stability of the enzyme are sensitive to various metal ions Mycolicibacterium neoaurum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.99.4 4-androstene-3,17-dione + acceptor Mycolicibacterium neoaurum
-
 1,4-androstadiene-3,17-dione + reduced acceptor
-
 ?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycolicibacterium neoaurum DSM 1381
-
 1,4-androstadiene-3,17-dione + reduced acceptor
-
 ?
1.3.99.4 a 3-oxosteroid + acceptor Mycolicibacterium neoaurum
-
 a 3-oxo-delta1-steroid + reduced acceptor
-
 ?
1.3.99.4 a 3-oxosteroid + acceptor Mycolicibacterium neoaurum DSM 1381
-
 a 3-oxo-delta1-steroid + reduced acceptor
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.99.4 Mycolicibacterium neoaurum A0A2P1IUZ5
-
-
1.3.99.4 Mycolicibacterium neoaurum DSM 1381 A0A2P1IUZ5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.99.4 recombinant N-terminally or C-terminally His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration Mycolicibacterium neoaurum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.99.4 1586
-
 purified enzyme, pH 8.0, 40°C, substrate 4-androstene-3,17-dione, conversion rate is 99% Mycolicibacterium neoaurum
1.3.99.4 1719.21
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 9alpha-hydroxy-4-androstene-3,17-dione Mycolicibacterium neoaurum
1.3.99.4 2560.15
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate canrenone Mycolicibacterium neoaurum
1.3.99.4 2623.11
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione Mycolicibacterium neoaurum
1.3.99.4 5975.86
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate androsta-4,9(11)-diene-3,17-dione Mycolicibacterium neoaurum
1.3.99.4 6299.29
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione acetate Mycolicibacterium neoaurum
1.3.99.4 7800.55
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 17alpha,21-dihydroxyprogesterone 21-acetate Mycolicibacterium neoaurum
1.3.99.4 8882.95
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 4-androstene-3,17-dione Mycolicibacterium neoaurum
1.3.99.4 9163.63
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 17alpha-hydroxypregn-4-ene-3,20-dione Mycolicibacterium neoaurum
1.3.99.4 12781.5
-
 pH 8.0, 40°C, purified recombinant enzyme, substrate 6,17alpha-epoxyprogesterone Mycolicibacterium neoaurum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.99.4 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione + acceptor hydrocortisone Mycolicibacterium neoaurum 1,4-pregnene-17alpha,21-diol-3,11,20-trione + reduced acceptor
-
 ?
1.3.99.4 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione + acceptor hydrocortisone Mycolicibacterium neoaurum DSM 1381 1,4-pregnene-17alpha,21-diol-3,11,20-trione + reduced acceptor
-
 ?
1.3.99.4 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione acetate + acceptor hydrocortisone acetate Mycolicibacterium neoaurum 1,4-pregnene-17alpha,21-diol-3,11,20-trione acetate + reduced acceptor
-
 ?
1.3.99.4 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione acetate + acceptor hydrocortisone acetate Mycolicibacterium neoaurum DSM 1381 1,4-pregnene-17alpha,21-diol-3,11,20-trione acetate + reduced acceptor
-
 ?
1.3.99.4 16,17alpha-epoxyprogesterone + acceptor
-
 Mycolicibacterium neoaurum ? + reduced acceptor
-
 ?
1.3.99.4 17alpha,21-dihydroxyprogesterone 21-acetate + acceptor RSA Mycolicibacterium neoaurum ? + reduced acceptor
-
 ?
1.3.99.4 17alpha-hydroxypregn-4-ene-3,20-dione + acceptor
-
 Mycolicibacterium neoaurum 1,4-pregnene-17alpha-ol-3,20-dione + reduced acceptor
-
 ?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
 Mycolicibacterium neoaurum 1,4-androstadiene-3,17-dione + reduced acceptor
-
 ?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
 Mycolicibacterium neoaurum DSM 1381 1,4-androstadiene-3,17-dione + reduced acceptor
-
 ?
1.3.99.4 9alpha-hydroxy-4-androstene-3,17-dione + acceptor
-
 Mycolicibacterium neoaurum ? + reduced acceptor
-
 ?
1.3.99.4 a 3-oxosteroid + acceptor
-
 Mycolicibacterium neoaurum a 3-oxo-delta1-steroid + reduced acceptor
-
 ?
1.3.99.4 a 3-oxosteroid + acceptor
-
 Mycolicibacterium neoaurum DSM 1381 a 3-oxo-delta1-steroid + reduced acceptor
-
 ?
1.3.99.4 androsta-4,9(11)-diene-3,17-dione + acceptor NSC 44826 Mycolicibacterium neoaurum ? + reduced acceptor
-
 ?
1.3.99.4 canrenone + acceptor
-
 Mycolicibacterium neoaurum ? + reduced acceptor
-
 ?
1.3.99.4 additional information enzyme KstD2 displays a high activity on a range of substrates, including 17alpha-hydroxypregn-4-ene-3,20-dione (17alpha-OH-P), androsta-4,9(11)-diene-3,17-dione (NSC 44826), and 4-androstene-3,17-dione (AD), substrate activity and kinetics, overview Mycolicibacterium neoaurum ?
-
-
1.3.99.4 additional information enzyme KstD2 displays a high activity on a range of substrates, including 17alpha-hydroxypregn-4-ene-3,20-dione (17alpha-OH-P), androsta-4,9(11)-diene-3,17-dione (NSC 44826), and 4-androstene-3,17-dione (AD), substrate activity and kinetics, overview Mycolicibacterium neoaurum DSM 1381 ?
-
-

Synonyms

EC Number Synonyms Comment Organism
1.3.99.4 3-ketosteroid-DELTA1-dehydrogenase
-
 Mycolicibacterium neoaurum
1.3.99.4 KSTD2
-
 Mycolicibacterium neoaurum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.99.4 40
-
-
 Mycolicibacterium neoaurum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.3.99.4 25 50 over 50% of maximal activity within this range Mycolicibacterium neoaurum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.3.99.4 30
-
 purified recombinant enzyme, completely stable at Mycolicibacterium neoaurum
1.3.99.4 40
-
 purified recombinant enzyme, 80% activity remaining Mycolicibacterium neoaurum
1.3.99.4 45
-
 purified recombinant enzyme, 20% activity remaining Mycolicibacterium neoaurum
1.3.99.4 50
-
 purified recombinant enzyme, inactivation Mycolicibacterium neoaurum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.99.4 1862.9
-
 9alpha-hydroxy-4-androstene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 2774.12
-
 canrenone recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 2842.34
-
 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 6475.3
-
 androsta-4,9(11)-diene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 6825.77
-
 11beta,17alpha,21-trihydroxypregn-4-ene-3,20-dione acetate recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 8452.5
-
 17alpha,21-dihydroxyprogesterone 21-acetate recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 9625.36
-
 4-androstene-3,17-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 9929.5
-
 17alpha-hydroxypregn-4-ene-3,20-dione recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum
1.3.99.4 13849.8
-
 16,17alpha-epoxyprogesterone recombinant enzyme, pH 8.0, 40°C Mycolicibacterium neoaurum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.99.4 8
-
-
 Mycolicibacterium neoaurum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.3.99.4 6 9 over 50% of maximal activity within this range Mycolicibacterium neoaurum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.3.99.4 5
-
 purified recombinant enzyme, 25% activity remaining Mycolicibacterium neoaurum
1.3.99.4 6 9 purified recombinant enzyme, over 80% activity remaining Mycolicibacterium neoaurum
1.3.99.4 10
-
 purified recombinant enzyme, 30% activity remaining Mycolicibacterium neoaurum

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.99.4 FAD
-
 Mycolicibacterium neoaurum

General Information

EC Number General Information Comment Organism
1.3.99.4 additional information homology-based structural analysis and structure modeling of KstD2 using the structure of SQ1-KstD1 Rhodococcus erythropolis SQ1 (PDB ID 4C3Y). Enzyme KstD1 and KstD2 belongs to clusters 1 and 2, respectively Mycolicibacterium neoaurum
1.3.99.4 physiological function 3-ketosteroid-DELTA1-dehydrogenase (KstD) catalyzes DELTA1-dehydrogenation and is involved in the steroid catabolism Mycolicibacterium neoaurum