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Literature summary extracted from

  • Kreit, J.
    Aerobic catabolism of sterols by microorganisms key enzymes that open the 3-ketosteroid nucleus (2019), FEMS Microbiol. Lett., 366, fnz173 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.3.99.4 O2 the DELTA1-dehydrogenation of 3-keto-4-en-steroid with KstD, purified from Nocardia corallina, is stimulated by molecular oxygen with stoichiometric production of hydrogen peroxide and 3-keto-1,4-diene-steroid Gordonia rubripertincta

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.99.4 gene acmB, recombinant expression in Escherichia coli Sterolibacterium denitrificans
1.3.99.4 gene kstD3, cloning and expression as a soluble protein in Escherichia coli Gordonia neofelifaecis

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 Gordonia rubripertincta
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Rhodococcus erythropolis
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Rhodococcus ruber
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Sterolibacterium denitrificans
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycolicibacterium smegmatis
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycobacterium tuberculosis
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Gordonia neofelifaecis
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Rhodococcus erythropolis SQ1
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycolicibacterium smegmatis ATCC 700084
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycobacterium tuberculosis H37Rv
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Gordonia neofelifaecis NRRL B-59395
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor Mycolicibacterium smegmatis mc(2)155
-
1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Gordonia rubripertincta
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Rhodococcus erythropolis
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Rhodococcus ruber
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Sterolibacterium denitrificans
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Mycolicibacterium smegmatis
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Mycobacterium tuberculosis
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Gordonia neofelifaecis
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Rhodococcus erythropolis SQ1
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Mycolicibacterium smegmatis ATCC 700084
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Mycobacterium tuberculosis H37Rv
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Gordonia neofelifaecis NRRL B-59395
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor Mycolicibacterium smegmatis mc(2)155
-
a 3-oxo-delta1-steroid + reduced acceptor
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.3.99.4 Gordonia neofelifaecis
-
five KstD isozymes
-
1.3.99.4 Gordonia neofelifaecis NRRL B-59395
-
five KstD isozymes
-
1.3.99.4 Gordonia rubripertincta
-
-
-
1.3.99.4 Mycobacterium tuberculosis P71864 a single enzyme
-
1.3.99.4 Mycobacterium tuberculosis H37Rv P71864 a single enzyme
-
1.3.99.4 Mycolicibacterium smegmatis A0R4S9 6 KstD isozymes
-
1.3.99.4 Mycolicibacterium smegmatis ATCC 700084 A0R4S9 6 KstD isozymes
-
1.3.99.4 Mycolicibacterium smegmatis mc(2)155 A0R4S9 6 KstD isozymes
-
1.3.99.4 Rhodococcus erythropolis Q9RA02 3 KstD isozymes
-
1.3.99.4 Rhodococcus erythropolis SQ1 Q9RA02 3 KstD isozymes
-
1.3.99.4 Rhodococcus ruber C6GCU2 three KstD isozymes
-
1.3.99.4 Rhodococcus ruber I0B6I6 three KstD isozymes
-
1.3.99.4 Rhodococcus ruber I0B6J0 three KstD isozymes
-
1.3.99.4 Sterolibacterium denitrificans A9XWD7 a single enzyme
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.99.4 , 4,9(11)-androstadiene-3,17-dione + acceptor
-
Gordonia neofelifaecis ? + reduced acceptor
-
?
1.3.99.4 , 4,9(11)-androstadiene-3,17-dione + acceptor
-
Gordonia neofelifaecis NRRL B-59395 ? + reduced acceptor
-
?
1.3.99.4 16alpha,17alpha-epoxyprogesterone + acceptor
-
Gordonia neofelifaecis ? + reduced acceptor
-
?
1.3.99.4 16alpha,17alpha-epoxyprogesterone + acceptor
-
Gordonia neofelifaecis NRRL B-59395 ? + reduced acceptor
-
?
1.3.99.4 25-hydroxycholest-4-en-3-one + acceptor
-
Sterolibacterium denitrificans ? + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Gordonia rubripertincta 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Rhodococcus erythropolis 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Rhodococcus ruber 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Sterolibacterium denitrificans 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Mycolicibacterium smegmatis 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Mycobacterium tuberculosis 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Gordonia neofelifaecis 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Rhodococcus erythropolis SQ1 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Mycolicibacterium smegmatis ATCC 700084 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Mycobacterium tuberculosis H37Rv 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Gordonia neofelifaecis NRRL B-59395 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-androstene-3,17-dione + acceptor
-
Mycolicibacterium smegmatis mc(2)155 1,4-androstadiene-3,17-dione + reduced acceptor
-
?
1.3.99.4 4-cholesten-3-one + acceptor
-
Sterolibacterium denitrificans ? + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Gordonia rubripertincta a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Rhodococcus erythropolis a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Rhodococcus ruber a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Sterolibacterium denitrificans a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Mycolicibacterium smegmatis a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Mycobacterium tuberculosis a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Gordonia neofelifaecis a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Rhodococcus erythropolis SQ1 a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Mycolicibacterium smegmatis ATCC 700084 a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Mycobacterium tuberculosis H37Rv a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Gordonia neofelifaecis NRRL B-59395 a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 a 3-oxosteroid + acceptor
-
Mycolicibacterium smegmatis mc(2)155 a 3-oxo-delta1-steroid + reduced acceptor
-
?
1.3.99.4 additional information KstD3 is active on a broad spectrum of substrates, comprising AD, progesterone, 4,9(11)-androstadiene-3,17-dione and 16alpha,17alpha-epoxyprogesterone. As usually the case with KstDs, the recombinant dehydrogenase is active neither with 3beta-/3alpha-hydroxysteroid nor with 3-ketosteroid containing a bulky C17-side chain, like 4-cholestene-3-one Gordonia neofelifaecis ?
-
-
1.3.99.4 additional information the purified KstD catalyzes hydrogen transfer from 3-keto-4-ene-steroid (donor) into 3-keto-1,4-diene-steroid (acceptor), e.g. progesterone to 1,4-androstadiene-3,17-dione (ADD). The transhydrogenation reaction reaches equilibrium, and obeys to a typical ping-pong mechanism in which the donor is the first substrate, while the acceptor is the second. The transhydrogenation turnover number is of the same magnitude as that of the C1,2-dehydrogenation Gordonia rubripertincta ?
-
-
1.3.99.4 additional information the recombinant enzyme is active with various 3-ketosteroids, including exceptionally the bulky side chain-containing steroids, 4-cholesten-3-one and 25-hydroxycholest-4-en-3-one Sterolibacterium denitrificans ?
-
-
1.3.99.4 additional information KstD3 is active on a broad spectrum of substrates, comprising AD, progesterone, 4,9(11)-androstadiene-3,17-dione and 16alpha,17alpha-epoxyprogesterone. As usually the case with KstDs, the recombinant dehydrogenase is active neither with 3beta-/3alpha-hydroxysteroid nor with 3-ketosteroid containing a bulky C17-side chain, like 4-cholestene-3-one Gordonia neofelifaecis NRRL B-59395 ?
-
-
1.3.99.4 progesterone + acceptor
-
Gordonia neofelifaecis ? + reduced acceptor
-
?

Subunits

EC Number Subunits Comment Organism
1.3.99.4 ? x * 62000, recombinant enzyme, SDS-PAGE Sterolibacterium denitrificans
1.3.99.4 ? x * 57000, recombinant KstD3, SDS-PAGE Gordonia neofelifaecis

Synonyms

EC Number Synonyms Comment Organism
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Gordonia rubripertincta
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Rhodococcus erythropolis
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Rhodococcus ruber
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Sterolibacterium denitrificans
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Mycolicibacterium smegmatis
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Mycobacterium tuberculosis
1.3.99.4 3-ketosteroid delta1-dehydrogenase
-
Gordonia neofelifaecis
1.3.99.4 AcmB
-
Sterolibacterium denitrificans
1.3.99.4 KstD
-
Gordonia rubripertincta
1.3.99.4 KstD
-
Rhodococcus erythropolis
1.3.99.4 KstD
-
Rhodococcus ruber
1.3.99.4 KstD
-
Sterolibacterium denitrificans
1.3.99.4 KstD
-
Mycolicibacterium smegmatis
1.3.99.4 KstD
-
Mycobacterium tuberculosis
1.3.99.4 KstD
-
Gordonia neofelifaecis
1.3.99.4 KSTD1
-
Rhodococcus ruber
1.3.99.4 KSTD2
-
Rhodococcus ruber
1.3.99.4 KSTD3
-
Rhodococcus ruber
1.3.99.4 Rv3537
-
Mycobacterium tuberculosis
1.3.99.4 Rv3537
-
Gordonia neofelifaecis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.99.4 8.4
-
transhydrogenation reaction Gordonia rubripertincta
1.3.99.4 8.5 10 DELTA1-dehydrogenation of 3-keto-4-en-steroid Gordonia rubripertincta
1.3.99.4 10
-
substrate C1,2-dehydrogenation, in presence of phenazine methosulfate (PMS) Gordonia rubripertincta

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.99.4 FAD
-
Sterolibacterium denitrificans
1.3.99.4 phenazine methosulfate PMS Gordonia rubripertincta

Expression

EC Number Organism Comment Expression
1.3.99.4 Rhodococcus erythropolis the enzyme is induced during the bacterial growth on 17alpha-methyl-testosterone up

General Information

EC Number General Information Comment Organism
1.3.99.4 evolution the genome of Rhodocccus ruber Chol-4 contains three genes that encode for 3-ketosteroid DELTA1-dehydrogenases: KstD1, KstD2 and KstD3. Their gene nucleotide sequences are similar to reciprocal homologues in other rhodococci. Physiological studies on strain Cho-4 and its kstD-deleted mutants demonstrate that the three dehydrogenases are involved in the catabolism of cholesterol at different degrees depending on catabolic intermediates. These KstDs are distinguished by their substrate profiles comprising C-19 and C-22 3-ketosteroids. Particularly, KstD3 has a preference for 5alpha-3-oxosteroids Rhodococcus ruber
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration. The DELTA1-dehydrogenation of 3-keto-4-en-steroid with KstD, purified from Nocardia corallina, is stimulated by molecular oxygen with stoichiometric production of hydrogen peroxide and 3-keto-1,4-diene-steroid. In addition, the purified KstD catalyzes hydrogen transfer from 3-keto-4-ene-steroid (donor) into 3-keto-1,4-dienesteroid (acceptor), e.g. progesterone to 1,4-androstadiene-3,17-dione (ADD). The purified KstD of N. corallina catalyzes efficiently the aromatization of A-ring of 19-nortestosterone and 19-norandrostenedione to produce respectively beta-estradiol and estrone (phenolic compounds). Also, 19-hydroxytestosterone, 19-hydroxyandrostenedione, and 19-oxotestosterone are reported to be substrates for this KstD. Their dehydrogenation produces the respective phenolic steroids. This steroid A-ring aromatization with the isolated KstD is similar to an earlier bioconversion, carried out using microbial cultures on 19-hydroxy-4-stene-3-one. The process of this bioconversion consists of the 19-hydroxystenone side chain cleavage first and second, the A-ring aromatization of the steroid derivative, producing beta-estradiol and/or estrone. Microbial 9alpha-hydroxylation does not occur in the process, due to the C19-hydroxyl group steric hindrance Gordonia rubripertincta
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Rhodococcus erythropolis
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Rhodococcus ruber
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Sterolibacterium denitrificans
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Mycolicibacterium smegmatis
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Mycobacterium tuberculosis
1.3.99.4 physiological function aerobic degradation of the sterol tetracyclic nucleus by microorganisms comprises the catabolism of A/B-rings, followed by that of C/D-rings. B-ring rupture at the C9,10-position is a key step involving 3-ketosteroid DELTA1-dehydrogenase (KstD) and 3-ketosteroid 9alpha-hydroxylase (KstH). Their activities lead to the aromatization of C4,5-en-containing A-ring causing the rupture of B-ring. C4,5alpha-hydrogenated 3-ketosteroid can be produced by the growing microorganism containing a 5alpha-reductase. In this case, the microorganism synthesizes, in addition to KstD and KstH, a 3-ketosteroid DELTA4-(5alpha)-dehydrogenase (Kst4D) in order to produce the A-ring aromatization, and consequently B-ring rupture. KstD and Kst4D are FAD-dependent oxidoreductases. KstH is composed of a reductase and a monooxygenase. This last component is the catalytic unit, it contains a Rieske-[2Fe-2S] center with a non-heme mononuclear iron in the active site. The characterized KstDs are active on 3-ketosteroids containing a short C17-chain or devoid of this chain. C-4,5-hydrogenated 3-ketosteroids are substrates, only if they are of the 5alpha-configuration Gordonia neofelifaecis