Human 20α-hydroxysteroid dehydrogenase (AKR1C1)-dependent biotransformation with recombinant fission yeast Schizosaccharomyces pombe

Naumann, Julia; Messinger, Josef; Bureik, Matthias

doi:10.1016/j.jbiotec.2010.08.004

Cited by 14 publications

(16 citation statements)

References 30 publications

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“…In large-scale biotransformations, whole-cell systems are often superior to the use of (partly) purified enzymes because enzyme expression as well as cofactor regeneration are done by the microbes themselves and scale-up of production is in principle only limited by the size of the available fermentation vessels. With respect to the choice of the host, recombinant fission yeasts of the species S. pombe have repeatedly been shown to be well suited for whole-cell biotransformation of steroids [6,22,[25][26][27][28][29] and for biotechnological production of both steroidal and non-steroidal drug metabolites [5,[30][31][32][33][34]. In a recent study, we described the first biotechnological use of human AKR1C1 (20α-HSD) heterologously expressed in S. pombe and demonstrated the reduction of progesterone as well as the synthetic progestogen dydrogesterone at the keto group at C20 position.…”

Section: Resultsmentioning

confidence: 99%

“…The AKR1C1 expression plasmid pREP1-AKR1C1 has already been described [6]. For the generation of strain CAD300 and CAD302, two further expression plasmids bearing the cDNA of human AKR1C1 were constructed.…”

Section: Akr1c1 Cdna and Vectorsmentioning

confidence: 99%

“…The AKR1C1 expression strain JMN8 has been described [6]. Fission yeast strain ATCC96115 with the genotype h − ade6-M210 leu1-32 ura4-D18 his3-Δ1 [12] was transformed using pREP1-AKR1C1 and pREP1ura-AKR1C1 to yield CAD300.…”

Section: Strains and Mediamentioning

confidence: 99%

“…Cells were generally centrifuged at 5,000×g at room temperature for 5 min. The medium used for fermentation was DM1 [6] containing 0.4% (w/v) glucose and 0.01% (w/v) adenin. The feeding medium was composed of 230 g L −1 glucose·H 2 O, 40 g L −1 (NH 4 ) 2 SO 4 , 10 g L −1 KH 2 PO 4 , 8 g L −1 MgSO 4 , 2 g L −1 Na 2 HPO 4 ·2H 2 O, 40 mL L −1 of salt stock solution, 2 mL L −1 of vitamin stock solution, and 200 μL L −1 of mineral stock solution supplemented by 2 g L −1 adenin.…”

Section: Strains and Mediamentioning

confidence: 99%

“…Recombinant fission yeast offers a great potential for the production of various enzymes of industrial interest, and its suitability for whole-cell biotransformations make this organism very promising for many biotechnological applications [4,5]. In a recent study, we demonstrated the first functional expression of a heterologous AKR in Schizosaccharomyces pombe using human AKR1C1 (20α-hydroxysteroid dehydrogenase) as the model enzyme [6]. We reported the general feasibility of a biotechnological use of this system for steroid modification and showed that the natural substrate progesterone as well as the synthetic progestogen dydrogesterone are regio-and stereospecifically reduced to 20α-dihydroprogesterone (20α-DHP) and 20α-dihydrodydrogesterone (20α-DHD), respectively.…”

Section: Introductionmentioning

confidence: 99%

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Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Naumann¹,

Zöllner²,

Drăgan³

et al. 2011

Appl Biochem Biotechnol

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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.

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Section: Resultsmentioning

confidence: 99%

Section: Akr1c1 Cdna and Vectorsmentioning

confidence: 99%

Section: Strains and Mediamentioning

confidence: 99%

Section: Strains and Mediamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Naumann¹,

Zöllner²,

Drăgan³

et al. 2011

Appl Biochem Biotechnol

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Humane Enzyme für die organische Synthese

et al. 2018

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Humane Enzyme wurden im Rahmen vieler Disziplinen untersucht. Die Anzahl an Reaktionen, die im menschlichen Körper stattfinden, ist enorm, ebenso wie die Anzahl möglicher Katalysatoren für eine Anwendung in der organischen Synthese. Dieser Aufsatz befasst sich mit chemischen Reaktionen humaner Enzyme, die eine Rolle im Metabolismus von Wirkstoffen oder Xenobiotika spielen. Einige dieser Reaktionen wurden im präparativen Maßstab untersucht. Der größte Teil an Anwendungen von humanen Enzymen stammt aus dem Bereich der Wirkstoffentwicklung, wo sie zur Synthese von Wirkstoffmetaboliten eingesetzt werden.

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Human Enzymes for Organic Synthesis

et al. 2018

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Human enzymes have been widely studied in various disciplines. The number of reactions taking place in the human body is vast, and so is the number of potential catalysts for synthesis. Herein, we focus on the application of human enzymes that catalyze chemical reactions in course of the metabolism of drugs and xenobiotics. Some of these reactions have been explored on the preparative scale. The major field of application of human enzymes is currently drug development, where they are applied for the synthesis of drug metabolites.

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Human 20α-hydroxysteroid dehydrogenase (AKR1C1)-dependent biotransformation with recombinant fission yeast Schizosaccharomyces pombe

Cited by 14 publications

References 30 publications

Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Humane Enzyme für die organische Synthese

Human Enzymes for Organic Synthesis

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