Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

Petrič, Špela; Hakki, Tarek; Bernhardt, Rita; Žigon, Dušan; Črešnar, Bronislava

doi:10.1016/j.jbiotec.2010.09.928

Cited by 60 publications

(76 citation statements)

References 43 publications

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“…After double enzyme-digestion of expression plasmid pCB1004-PgpdA-CPR with Bam HI/Hind III, there were two bright bands at 2027 bp and 6557 bp. The DNA fragments were recovered and sequenced to confirm the results, which were consistent with the reported sequences [18].…”

Section: Expression Plasmid Identificationsupporting

confidence: 78%

“…Therefore, the cloning of CPR has been becoming very important for the biotransformation reaction of hydroxylation. Some reports described the cloning of CPR DNA from mammals, such as rat [7][8][9], guinea pig [10], mouse [11], rabbit [12], human [13], hamster [14], pig [15], horse [16], koala [17], and the microorganism, Rhizopus oryzae [18]. Some of cloned CPR genes were expressed in the bacteria or yeasts, but never in the filamentous fungi, Rhizopus nigricans.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular cloning, expression of CPR gene from Rhizopus oryzae into Rhizopus nigericans and its application in the 11α-hydroxylation of 16α, 17-epoxy-progesterone

Chen

Cao

et al. 2014

Enzyme and Microbial Technology

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Section: Expression Plasmid Identificationsupporting

confidence: 78%

Section: Introductionmentioning

confidence: 99%

Molecular cloning, expression of CPR gene from Rhizopus oryzae into Rhizopus nigericans and its application in the 11α-hydroxylation of 16α, 17-epoxy-progesterone

Chen

Cao

et al. 2014

Enzyme and Microbial Technology

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“…For many of these reactions, chemical synthesis is tedious, and natural microorganisms that permit the respective biotransformations are not always known or applicable. Hence, the use of recombinant microorganisms is an economically reasonable alternative [19] that has so far mainly focused on cytochrome P450 (CYP) enzymes [20][21][22][23][24]. Although some enzymes of the AKR superfamily are also promising for specific steroid modifications, they have not yet been applied in industrial scale.…”

Section: Resultsmentioning

confidence: 99%

“…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%

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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“…and Aspergillus spp. are able to carry out 11α-hydroxylations (Petrič et al 2010;Samanta and Ghosh 1987), Curvularia spp. and Cunninghamella spp.…”

Section: Biotransformation Of Steroids By Fungimentioning

confidence: 99%

Fifty years of drug discovery from fungi

2011

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For the past 50 years, fungal secondary metabolites have revolutionized medicine yielding blockbuster drugs and drug leads of enormous therapeutic and agricultural potential. Since the discovery of penicillin, the first β-lactam antibiotic, fungi provided modern medicine with important antibiotics for curing life threatening infectious diseases. A new era in immunopharmacology and organ transplantation began with the discovery of cyclosporine. Other important drugs or products for agriculture derived from or inspired by natural products from fungi include statins, echinocandins and strobilurins. Moreover, fungal biotransformation of steroids for the industrial production of steroidal hormones represents one of the key successes in biotechnology. Given that estimations of fungal biodiversity exceed by far the number of already identified species, chances to find hitherto unidentified fungal species and novel bioactive fungal products are still high. Thus, further compounds with medicinal or agricultural potential from less investigated fungal taxa can be expected in the years to come.

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Discovery of a steroid 11α-hydroxylase from Rhizopus oryzae and its biotechnological application

Cited by 60 publications

References 43 publications

Molecular cloning, expression of CPR gene from Rhizopus oryzae into Rhizopus nigericans and its application in the 11α-hydroxylation of 16α, 17-epoxy-progesterone

Molecular cloning, expression of CPR gene from Rhizopus oryzae into Rhizopus nigericans and its application in the 11α-hydroxylation of 16α, 17-epoxy-progesterone

Biotechnological Production of 20-alpha-Dihydrodydrogesterone at Pilot Scale

Fifty years of drug discovery from fungi

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