Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Abstract

The human sex hormone progesterone plays an essential and complex role in a number of physiological processes. Progesterone deficiency is associated with menstrual disorders and infertility as well as premature birth and abortion. For progesterone replacement therapy, the synthetic progestogen dydrogesterone is commonly used. In the body, this drug is metabolized to 20α-dihydrodydrogesterone (20α-DHD), which also shows extensive pharmacological effects and hence could act as a therapeutic agent itself. In this study, we describe an efficient biotechnological production procedure for 20α-DHD that employs the stereo- and regioselective reduction of dydrogesterone in a whole-cell biotransformation process based on recombinant fission yeast cells expressing the human enzyme AKR1C1 (20α-hydroxysteroid dehydrogenase, 20α-HSD). In a fed-batch fermentation at pilot scale (70 L) with a genetically improved production strain and under optimized reaction conditions, an average 20α-DHD production rate of 190 μM day(-1) was determined for a total biotransformation time of 136 h. Combined with an effective and reliable downstream processing, a continuous production rate of 12.3 ± 1.4 g 20α-DHD per week and fermenter was achieved. We thus established an AKR-dependent whole-cell biotransformation process that can also be used for the production of other AKR1C1 substrates (as exemplarily shown by the production of 20α-dihydroprogesterone in gram scale) and is in principle suited for the production of further human AKR metabolites at industrial scale.

Expression of human aldo-keto reductase 1C2 in cell lines of peritoneal endometriosis: Potential implications in metabolism of progesterone and dydrogesterone and inhibition by progestins

“…The 1 H NMR spectrum was recorded on a Bruker Advance 400 DPX spectrometer at 302 K using solvent as an internal standard (CDCl 3 at 7.26 ppm). In the spectrum the signals that unambiguously prove the identity of the produced 20␣-dihydrodydrogesterone (the chemical shifts are identical to those published previously [26]) are clearly visible and are broadened in Fig. 4.…”

Yeast as a promising heterologous host for steroid bioproduction

“…As discussed in this session, different yeast strains have been utilized for the production of various steroids through biotransformation strategy and generally with good conversion efficiency. The medium condition of steroid biotransformation in yeast is relatively simple, usually common yeast rich or minimal medium supplemented with organic solvent, such as ethanol and N,N-dimethylformamide (DMF) to help dissolve the steroid substrates [48,56]. Addition of β-cyclodextrin often helps the transport of steroid substrates into the cells [50,57].…”

Microwave‐assisted synthesis of novel steroidal heterocyclic analogs as potent inhibitors of RANKL‐induced osteoclastogenesis

“…Steroids are polycyclic compounds that exhibit various biological activities and therefore can be transformed into medications to treat a variety of ailments (Naumann et al, 2011). Pregnenolone I , an endogenous steroid that occurs naturally, is the biosynthetic precursor of many steroidal hormones such as estrogen, progesterone, testosterone, glucocorticoids, and mineralocorticoids (Miller & Auchus, 2011; Schiffer et al, 2019).…”

“…Steroids are polycyclic compounds that exhibit various biological activities and therefore can be transformed into medications to treat a variety of ailments (Naumann et al, 2011).…”