Abstract
Microbiological synthesis of 7α- and 7β-hydroxy derivatives of testololactone and testolactone was developed based on bioconversion of dehydroepiandrosterone (DHEA) by fungus of Isaria fumosorosea VKM F-881 with subsequent modification of the obtained stereoisomers by actinobacteria. The first stage included obtaining of the stereoisomers of 3β,7(α/β)-dihydroxy-17a-oxa-D-homo-androst-5-en-17-ones in the preparative amounts. Then the conversion of 7-hydroxylated D-lactones obtained by selected actinobacteria of Nocardioides simplex VKM Ac-2033D, Saccharopolyspora hirsuta VKM Ac-666, and Streptomyces parvulus MTOC Ac-21v was studied. Under the transformation of 3β,7α-dihydroxy-17a-oxa-D-homo-androst-5-en-17-one and its corresponding 7β-stereoisomer by N. simplex VKM Ac-2033D and S. hirsuta VKM Ac-666 the 7α- and 7β-hydroxy-17a-oxa-D-homo-androst-4-ene-3,17-dione (7α- and 7β-hydroxytestololactone), 7α- and 7β-hydroxy-17a-oxa-D-homo-androsta-1,4-diene-3,17-dione (7α- and 7β-hydroxytestolactone) were obtained with molar yields in a range of 60.3–90.9 mol%. The crystalline products of 7α-hydroxytestololactone, 7α-hydroxytestolactone, and their corresponding 7β-hydroxy stereoisomers were isolated, and their structures were confirmed by mass spectrometry and 1H-NMR spectroscopy analyses. The strain of Str. parvulus MTOC Ac-21v transformed 3β,7(α/β)-dihydroxy-17a-oxa-D-homo-androst-5-en-17-ones into the corresponding 3-keto-4-ene analogs and did not show 3-ketosteroid 1(2)-dehydrogenase activity. The activity of actinobacteria towards steroid D-lactones was hitherto unreported.
The results contribute to the knowledge of metabolic versatility of actinobacteria capable of transforming steroid substrates and may be applied in the synthesis of potential aromatase inhibitors.
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Acknowledgments
Authors are grateful to thank Ph.D. I. S. Levina (N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences) for mass spectrometric and 1H-NMR analysis.
Funding
The research was carried out within the State Assignment (No. 0114-2018-0004). The work was supported by Russian Science Foundation (No. 18-14-00361).
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Lobastova, T.G., Khomutov, S.M., Shutov, A.A. et al. Microbiological synthesis of stereoisomeric 7(α/β)-hydroxytestololactones and 7(α/β)-hydroxytestolactones. Appl Microbiol Biotechnol 103, 4967–4976 (2019). https://doi.org/10.1007/s00253-019-09828-6
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DOI: https://doi.org/10.1007/s00253-019-09828-6