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Literature summary for 1.3.99.4 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)

Cloned (Comment) Organism
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

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Mycolicibacterium neoaurum A0A2P1IUZ5
-
-
Mycolicibacterium neoaurum DSM 1381 A0A2P1IUZ5
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
?
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
-
?
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
-
?
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
-
?
16,17alpha-epoxyprogesterone + acceptor
-
Mycolicibacterium neoaurum ? + reduced acceptor
-
?
17alpha,21-dihydroxyprogesterone 21-acetate + acceptor RSA Mycolicibacterium neoaurum ? + reduced acceptor
-
?
17alpha-hydroxypregn-4-ene-3,20-dione + acceptor
-
Mycolicibacterium neoaurum 1,4-pregnene-17alpha-ol-3,20-dione + reduced acceptor
-
?
4-androstene-3,17-dione + acceptor
-
Mycolicibacterium neoaurum 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
4-androstene-3,17-dione + acceptor
-
Mycolicibacterium neoaurum DSM 1381 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
9alpha-hydroxy-4-androstene-3,17-dione + acceptor
-
Mycolicibacterium neoaurum ? + reduced acceptor
-
?
a 3-oxosteroid + acceptor
-
Mycolicibacterium neoaurum a 3-oxo-delta1-steroid + reduced acceptor
-
?
a 3-oxosteroid + acceptor
-
Mycolicibacterium neoaurum DSM 1381 a 3-oxo-delta1-steroid + reduced acceptor
-
?
androsta-4,9(11)-diene-3,17-dione + acceptor NSC 44826 Mycolicibacterium neoaurum ? + reduced acceptor
-
?
canrenone + acceptor
-
Mycolicibacterium neoaurum ? + reduced acceptor
-
?
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 ?
-
-
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

Synonyms Comment Organism
3-ketosteroid-DELTA1-dehydrogenase
-
Mycolicibacterium neoaurum
KSTD2
-
Mycolicibacterium neoaurum

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Mycolicibacterium neoaurum

pH Range

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

pH Stability

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

Cofactor

Cofactor Comment Organism Structure
FAD
-
Mycolicibacterium neoaurum

General Information

General Information Comment Organism
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
physiological function 3-ketosteroid-DELTA1-dehydrogenase (KstD) catalyzes DELTA1-dehydrogenation and is involved in the steroid catabolism Mycolicibacterium neoaurum