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Metab Eng. 2016 Jul;36:19-27. doi: 10.1016/j.ymben.2016.02.010. Epub 2016 Mar 11.

Genetic engineering of Bacillus megaterium for high-yield production of the major teleost progestogens 17α,20β-di- and 17α,20β,21α-trihydroxy-4-pregnen-3-one.

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Saarland University, Institute of Biochemistry, Campus B2.2, 66123 Saarbrücken, Germany.
Saarland University, Institute of Pharmaceutical Biology, Campus C2.3, 66123 Saarbrücken, Germany.
Saarland University, Institute of Biochemistry, Campus B2.2, 66123 Saarbrücken, Germany. Electronic address:


17α,20β-Dihydroxy-4-pregnen-3-one (17α,20βDiOH-P) and 17α,20β,21α-trihydroxy-4-pregnen-3-one (20βOH-RSS) are the critical hormones required for oocyte maturation in fish. We utilized B. megaterium's endogenous 20β-hydroxysteroid dehydrogenase (20βHSD) for the efficient production of both progestogens after genetically modifying the microorganism to reduce side-product formation. First, the gene encoding the autologous cytochrome P450 CYP106A1 was deleted, resulting in a strain devoid of any steroid hydroxylation activity. Cultivation of this strain in the presence of 17α-hydroxyprogesterone (17αOH-P) led to the formation of 17α,20α-dihydroxy-4-pregnen-3-one (17α,20αDiOH-P) as a major and 17α,20βDiOH-P as a minor product. Four enzymes were identified as 20αHSDs and their genes deleted to yield a strain with no 20αHSD activity. The 3-oxoacyl-(acyl-carrier-protein) reductase FabG was found to exhibit 20βHSD-activity and overexpressed to create a biocatalyst yielding 0.22g/L 17α,20βDiOH-P and 0.34g/L 20βOH-RSS after 8h using shake-flask cultivation, thus obtaining products that are at least a thousand times more expensive than their substrates.


17α,20β,21α-Trihydroxy-4-pregnen-3-one; 17α,20β-Dihydroxy-4-pregnen-3-one; Bacillus megaterium; Hydroxysteroid dehydrogenase; MIH; Steroid hormones

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