Bioconversion of steroids by Cochliobolus lunatus—II. 11β-hydroxylation of 17α,21-dihydroxypregna-1,4-diene-3,20-dione 17-acetate in dependence of the inducer structure

Undisz, Katrin; Groh, Helmuth; Stopsack, H.; Hörhold‐Schubert, Cläre

doi:10.1016/0960-0760(92)90242-b

Cited by 17 publications

(3 citation statements)

References 9 publications

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“…Fungal CYPs are usually induced by their substrates (Undisz et al ., ), and thus we decided to analyse by RT‐PCR the relative mRNA levels of the selected CYP and CPR encoding genes under induction conditions. C. lunatus mycelium was induced in the presence of DOC, progesterone and androst‐4‐ene‐3,17‐dione (AD) while cholic acid and β‐sitosterol were used as negative control for non‐induced conditions (Undisz et al ., ). The genes encoding three of the selected eight putative CYPs (CYP115117, CYP34615 and CYP56034) were not expressed neither under induced or non‐induced conditions (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Biochemical studies of the 11β‐hydroxylase of C. lunatus described it as a microsomal two‐component monooxygenase system which is composed of a CYP and a NADPH‐CPR (Janig et al ., ; Suzuki et al ., ). The 11β‐hydroxylase can be specifically induced by some steroids (Undisz et al ., ). On the other hand, two CPRs named CPR1 (CPR64795) and CPR2 (CPR128465) had been previously characterized in C. lunatus (Lah et al ., , ) and both enzymes have been expressed in Escherichia coli and reconstituted in vitro with the CYP53A15 of C. lunatus having a benzoate 4‐hydroxylase activity.…”

Section: Discussionmentioning

confidence: 97%

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Identification and expression of the 11β‐steroid hydroxylase from Cochliobolus lunatus in Corynebacterium glutamicum

Felpeto-Santero

Galán

Luengo

et al. 2019

Microbial Biotechnology

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Summary Hydroxylation of steroids has acquired special relevance for the pharmaceutical industries. Particularly, the 11β‐hydroxylation of steroids is a reaction of biotechnological importance currently carried out at industrial scale by the fungus Cochliobolus lunatus. In this work, we have identified the genes encoding the cytochrome CYP103168 and the reductase CPR64795 of C. lunatus responsible for the 11β‐hydroxylase activity in this fungus, which is the key step for the preparative synthesis of cortisol in industry. A recombinant Corynebacterium glutamicum strain harbouring a plasmid expressing both genes forming a synthetic bacterial operon was able to 11β‐hydroxylate several steroids as substrates. This is a new example to show that the industrial strain C. glutamicum can be used as a suitable chassis to perform steroid biotransformation expressing eukaryotic cytochromes.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 97%

Identification and expression of the 11β‐steroid hydroxylase from Cochliobolus lunatus in Corynebacterium glutamicum

Felpeto-Santero

Galán

Luengo

et al. 2019

Microbial Biotechnology

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“…These hydroxylation sites are distinct from those previously described for Δ 4-5 steroids in reactions with the same fungus, that usually occur at the positions 11β, 14α and 7α. 7,23,24 Such differences may be explained by distinct enzyme/ substrate interactions, arising from the uncommon configuration of digitoxigenin steroidal frame (cis, trans, cis), in comparison to other steroids, or due to the presence of the α,β-unsaturated lactone ring at C-17. Such hypotheses consider that the same hydroxylases do participate in reactions with digitoxigenin and other steroids.…”

Section: Resultsmentioning

confidence: 99%

Biotransformation of digitoxigenin by Cochliobolus lunatus

Pádua¹,

Oliveira²,

Filho³

et al. 2007

J. Braz. Chem. Soc.

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A reação de biotransformação da digitoxigenina (1) por Cochliobolus lunatus foi investigada. Foram realizados experimentos com duração de 4 dias, que resultaram no isolamento de quatro produtos, cujas estruturas químicas foram elucidadas como sendo 1β-hidroxidigitoxigenina (2), 7β-hidroxidigitoxigenina (3), 8β-hidroxidigitoxigenina (4) e digitoxigenona (5). A obtenção desses produtos nas condições empregadas nunca foi anteriormente descrita. A produção da substância 4 em uma reação de biotransformação também é inédita.The biotransformation of digitoxigenin (1) by Cochliobolus lunatus was investigated. The biotransformation reaction was carried out in a 4-day process, resulting in the isolation of four products, whose structures were elucidated as 1β-hydroxydigitoxigenin (2), 7β-hydroxydigitoxigenin (3), 8β-hydroxydigitoxigenin (4) and digitoxigenone (5). The production of these derivatives under the employed conditions has never been described so far. This is also the first report on the production of compound 4 by a biotransformation reaction.Keywords: digitoxigenin, Cochliobolus lunatus, 1β-hydroxydigitoxigenin, 7β-hydroxydigitoxigenin, 8β-hydroxydigitoxigenin, digitoxigenone IntroductionEnzymes are known to possess a wide substrate tolerance by keeping their exquisite catalytic properties with respect to chemo-, regio-and enantio-selectivity, playing an important role in biotransformations. 1 Biotransformation reactions can be accomplished at room temperature and in aqueous medium, presenting itself as a milder alternative to classical chemical reactions, 2-4 being employed for the resolution of racemates and to introduce chiral centers in substrates, among other uses. 5 Fungi are eukaryotic organisms that possess enzyme systems similar to those of mammalians. They usually present highly flexible metabolism, thus accepting varied sources of carbon and nitrogen. The ecological relations of these organisms include the metabolism of different secondary compounds, a feature sustained by diversified enzymatic systems, both intra and extra cellular, capable of carrying out numerous reactions. 6-8 Such attributes suggest fungi as suitable organisms to perform biotransformation reactions.The biotransformation of steroidal compounds by fungi has been extensively evaluated, including reactions with cardiac glycosides (Pádua et al. 9 and references herein; Table 3 of the present work). Digoxin, a Digitalis cardenolide, is still the drug of choice for the treatment of congestive heart failure, acting as a selective inhibitor of the Na + ,K + ATPase enzyme. Biotransformation of cardenolides has been investigated either as a strategy to obtain new derivatives or to convert the A-type cardenolides into the corresponding C-type compounds, which have clinical relevance. 9 The main reactions obtained so far for cardenolide biotransformation were hydroxylation in different positions of the steroidal skeleton, oxidation, glycosylation, epimerization and esterification of the hydroxyl group at C-3 (see Table 3 for references). An...

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Steroid Bioconversion

Fernandes¹,

Cabral²

2010

Encyclopedia of Industrial Biotechnology

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Steroids constitute a particular class of lipids characterized by a typical tetracyclic skeleton, composed of one five‐member ring and three fused six‐member rings. Steroids are widely used as therapeutic agents and since their inception in the market, research efforts have been made in order to improve production processes as well as to develop novel synthetic molecules, with enhanced efficiency and reduced side effects. Given the complexity of the steroid skeleton, total chemical synthesis of given steroid molecules is hardly an effective approach; hence, steroid production processes rely on chemical modifications of educts that structurally resemble the targeted product. Still, steroid molecules present multiple chiral centers, a feature that makes particularly appealing the use of the selective microbial catalysts to provide a pathway that does not require protection, and additional deprotection steps. Furthermore, bioconversions are performed in a milder, greener environment, as compared to the purely chemical approach. The present work aims to provide an overview of steroid bioconversions. The potential of biologic agents (microbial cells/enzymes) to catalyze the conversion of a wide array of steroid educts will be presented, with particular emphasis on those that have achieved particular relevance, as well as on key aspects within the field. The chapter ends with the perspective trends in the field of steroid bioconversions

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Bioconversion of steroids by Cochliobolus lunatus—II. 11β-hydroxylation of 17α,21-dihydroxypregna-1,4-diene-3,20-dione 17-acetate in dependence of the inducer structure

Cited by 17 publications

References 9 publications

Identification and expression of the 11β‐steroid hydroxylase from Cochliobolus lunatus in Corynebacterium glutamicum

Identification and expression of the 11β‐steroid hydroxylase from Cochliobolus lunatus in Corynebacterium glutamicum

Biotransformation of digitoxigenin by Cochliobolus lunatus

Steroid Bioconversion

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