Microbiological Transformations. IX. The 1β-Hydroxylation of Androstenedione

Dodson, R. M.; Kraychy, Stephen; Nicholson, Robert T.; Mizuba, Seth

doi:10.1021/jo01056a043

Cited by 20 publications

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“…The 1 H NMR spectrum of IV showed a singlet at d 4.64, corresponding to a carbon signal resonating at d 73.4 (C-6) in the 13 C NMR spectrum. Comparing with the published data (Hanson et al 1996;Dodson et al 1962) this indicated the position of hydroxyl group to be C-6 with a b-orientation. Compound V had a molecular formula of C 19 H 24 O 3 , deduced from its mass spectrum, it contained 16 units more than ADD.…”

Section: Resultssupporting

confidence: 63%

Microbiological hydroxylation of androst-1,4-dien-3,17-dione byNeurospora crassa

Faramarzi¹,

Hajarolasvadi²,

Yazdi³

et al. 2007

Biocatalysis and Biotransformation

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Nine hydroxy-derived androstadiene compounds were isolated from the fermentation broth of Neurospora crassa when incubated in the presence of androst-1,4-dien-3,17-dione (ADD; I) for 7 days. Hydroxylations at 6b, 7b, 11a, 14a-positions and 17-carbonyl reduction of the substrate were the characteristics observed in this biotransformation. Their structures were determined by spectroscopic methods as 17b-hydroxyandrost-1,4-dien-3-one (II), 14a-hydroxyandrost-1,4-dien-3,17-dione (III), 6b-hydroxyandrost-1,4-dien-3,17-dione (IV), 11a-hydroxyandrost-1,4-dien-3,17-dione (V), 6b,17b-dihydroxyandrost-1,4-dien-3-one (VI), 7b-hydroxyandrost-1,4-dien-3,17-dione (VII), 14a,17b-dihydroxyandrost-1,4-dien-3-one (VIII), 6b,14a-dihydroxyandrost-1,4-dien-3,17-dione (IX), and 11a,17b-dihydroxyandrost-1,4-dien-3-one (X). A new steroid substance, 6b,14a-dihydroxyandrost-1,4-dien-3,17-dione (IX), was also characterized during this study. The best fermentation condition was found to be 7-day incubation at 258C and pH values of 5.0 Á6.0 in the presence of 0.05 g 100 mL (1 of the substrate. At a concentration above 0.075 g 100 mL (1 , the biotransformation was completely inhibited.

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Section: Resultssupporting

confidence: 63%

Microbiological hydroxylation of androst-1,4-dien-3,17-dione byNeurospora crassa

Faramarzi¹,

Hajarolasvadi²,

Yazdi³

et al. 2007

Biocatalysis and Biotransformation

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“…Other work with the 1β‐hydroxytestosterone derived from Xyleria oxidations [34] has yielded reports that it is a weak inhibitor of human placental aromatase (P450 19A1), the enzyme that oxidizes testosterone to 17β‐estradiol, with an IC 50 value of ≈ 1 m m [38]. Another report indicated that human placental microsomes used 1β‐hydroxytestosterone ≈ 30% as efficiently as testosterone or antrostenedione [35], but apparently has not been confirmed.…”

Section: Discussionmentioning

confidence: 99%

“…Porcine gonadal P450 19A1 (aromatase) converted testosterone to significant amounts of 1β‐hydroxytestosterone, as well as 19‐hydroxy‐ and 19‐oxotestosterone and 17β‐estradiol [39]. The assignment of the structure was based on (a) comparison of an MS fragmentation pattern with an earlier literature spectrum [35] (going back to the original Xylaria product [34]), and (b) labilization of 3 H from [1β‐ 3 H]‐testosterone [39]. Corbin et al .…”

Section: Discussionmentioning

confidence: 99%

“…The 1(b)-hydroxylation of androgens has been reported previously. Dodson et al [34] reported that microbial (Xylaria sp.) oxidation of androstenedione yielded a product identified as the 1b-hydroxy derivative.…”

Section: Discussionmentioning

confidence: 99%

“…(going back to the original Xylaria product[34]), and (b) labilization of 3 H from [1b-3 H]-testosterone[39]. Corbin NMR shifts (see.…”

mentioning

confidence: 99%

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Testosterone 1β‐hydroxylation by human cytochrome P450 3A4

Krauser

Voehler

Tseng

et al. 2004

European Journal of Biochemistry

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Human cytochrome P450 3A4 forms a series of minor testosterone hydroxylation products in addition to 6b-hydroxytestosterone, the major product. One of these, formed at the next highest rate after the 6b-and 2b-hydroxy products, was identified as 1b-hydroxytestosterone. This product was characterized from a mixture of testosterone oxidation products using an HPLC-solid phase extraction-cryoprobe NMR/time-of-flight mass spectrometry system, with an estimated total of 6 lg of this product. Mass spectrometry established the formula as C 19 H 29 O 3 (MH + 305.2080). The 1-position of the added hydroxyl group was established by correlated spectroscopy and heteronuclear spin quantum correlation experiments, and the b-stereochemistry of the added hydroxyl group was assigned with a nuclear Overhauser correlated spectroscopy experiment (1a-H). Of several human P450s examined, only P450 3A4 formed this product. The product was also formed in human liver microsomes.Keywords: cytochrome P450; NMR spectroscopy; HPLC-NMR combinations; testosterone.Cytochrome P450 (P450; also termed heme-thiolate P450 [1]) enzymes have long been of interest because of their roles in steroid metabolism [2,3]. These oxidations are most critical in steroidogenic tissues, and a set of 12 P450s are most important [4,5]. The hepatic P450s have also been studied extensively in the context of their abilities to hydroxylate steroids, even though few of the oxidations involve the generation of products with distinctive biological activities. Seminal in this area is the work of Conney and his associates, who studied the hydroxylation of testosterone in rat liver systems and developed the hypothesis that different hydroxylations are catalyzed by individual P450 enzymes [6]. Subsequently testosterone hydroxylation patterns have been utilized extensively as probes of the presence and function of individual rat liver P450s [7][8][9][10][11].Testosterone hydroxylation has also been studied extensively with human liver microsomal P450s. Early work with liver microsomes resulted in reports of hydroxylation at the 2b, 6a, 6b, 7a, 15b, 16a, and 17 positions (17-hydroxylation yields the ketone androstenedione) [12][13][14][15][16][17]. A human liver P450 was isolated that was shown to be the major 6b-hydroxylase [18]; this P450 was originally termed nifedipine oxidase (P450 NF ) and subsequently named P450 3A4. Other work confirmed the role of P450 3A4 as the major enzyme involved in testosterone 6b-hydroxylation [19]. Other P450 3A subfamily enzymes (3A5, 3A7, 3A43) can also catalyze this reaction [20,21].Testosterone hydroxylation is in general use today as one of the characteristic assays of P450 3A4, which was subsequently shown to be the most abundant P450 in human liver and small intestine [22] and involved in the oxidation of approximately one-half the drugs used today [23]. The major product is 6b-hydroxytestosterone [18,19] but several other hydroxylations occur, including those at the 2b and 15b positions [18,19] (Fig. 1). Not all of the products have bee...

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Substrate Hunting for the Myxobacterial CYP260A1 Revealed New 1α‐Hydroxylated Products from C‐19 Steroids

et al. 2015

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Cytochromes P450 catalyze a variety of synthetically useful reactions. However, it is difficult to determine their physiological or artificial functions when a plethora of orphan P450 systems are present in a genome. CYP260A1 from Sorangium cellulosum So ce56 is a new member among the 21 available P450s in the strain. To identify putative substrates for CYP260A1 we used high-throughput screening of a compound library (ca. 17,000 ligands). Structural analogues of the type I hits were searched for biotechnologically relevant compounds, and this led us to select C-19 steroids as potential substrates. We identified efficient surrogate redox partners for CYP260A1, and an Escherichia coli-based whole-cell biocatalyst system was developed to convert testosterone, androstenedione, and their derivatives methyltestosterone and 11-oxoandrostenedione. A detailed (1) H and (13) C NMR characterization of the product(s) from C-19 steroids revealed that CYP260A1 is the very first 1α-steroid hydroxylase.

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Microbiological Transformations. IX. The 1β-Hydroxylation of Androstenedione

Cited by 20 publications

References 0 publications

Microbiological hydroxylation of androst-1,4-dien-3,17-dione byNeurospora crassa

Microbiological hydroxylation of androst-1,4-dien-3,17-dione byNeurospora crassa

Testosterone 1β‐hydroxylation by human cytochrome P450 3A4

Substrate Hunting for the Myxobacterial CYP260A1 Revealed New 1α‐Hydroxylated Products from C‐19 Steroids

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