Hydroxylation of lithocholic acid by selected actinobacteria and filamentous fungi

Kollerov, Vyacheslav; Monti, Daniela; Deshcherevskaya, N.O.; Lobastova, Tatyana G.; Ferrandi, Erica Elisa; Larovere, A.; Gulevskaya, S. A.; Riva, Sergio; Donova, Marina V.

doi:10.1016/j.steroids.2012.12.010

Cited by 25 publications

(23 citation statements)

References 28 publications

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“…7α-hydroxylase activity towards lithocholic (LCA) and deoxycholic (DCA) acids was described for Actinobacteria of Amycolatopsis , Lentzea , Pseudonocardia , and Saccharopolyspora genera. New strains capable of 7α-hydroxylation of DCA were revealed also among Nocardiopsis , Nonomuraea , and Saccharothrix species (Deshcherevskaya et al 2016 ; Kollerov et al 2013 ). The reactions are of importance for biotechnological production of valuable cholic acids (e.g., ursodeoxycholic, ursocholic acids).…”

Section: Introductionmentioning

confidence: 99%

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Szaleniec

Wojtkiewicz

Bernhardt

et al. 2018

Appl Microbiol Biotechnol

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The steroid superfamily includes a wide range of compounds that are essential for living organisms of the animal and plant kingdoms. Structural modifications of steroids highly affect their biological activity. In this review, we focus on hydroxylation of steroids by bacterial hydroxylases, which take part in steroid catabolic pathways and play an important role in steroid degradation. We compare three distinct classes of metalloenzymes responsible for aerobic or anaerobic hydroxylation of steroids, namely: cytochrome P450, Rieske-type monooxygenase 3-ketosteroid 9α-hydroxylase, and molybdenum-containing steroid C25 dehydrogenases. We analyze the available literature data on reactivity, regioselectivity, and potential application of these enzymes in organic synthesis of hydroxysteroids. Moreover, we describe mechanistic hypotheses proposed for all three classes of enzymes along with experimental and theoretical evidences, which have provided grounds for their formulation. In case of the 3-ketosteroid 9α-hydroxylase, such a mechanistic hypothesis is formulated for the first time in the literature based on studies conducted for other Rieske monooxygenases. Finally, we provide comparative analysis of similarities and differences in the reaction mechanisms utilized by bacterial steroid hydroxylases.

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Section: Introductionmentioning

confidence: 99%

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Szaleniec

Wojtkiewicz

Bernhardt

et al. 2018

Appl Microbiol Biotechnol

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“…Later, many other microorganisms with a 7β-hydroxylating activity were discovered in strains of actinobacteria and ﬁlamentous fungi [ 96 – 97 ]. The key-enzyme in that pathway is a P450-like enzyme that catalyses the specific and irreversible 7β-hydroxylation.…”

Section: Reviewmentioning

confidence: 99%

Latest development in the synthesis of ursodeoxycholic acid (UDCA): a critical review

Tonin

Arends

2018

Beilstein J. Org. Chem.

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Ursodeoxycholic acid (UDCA) is a pharmaceutical ingredient widely used in clinics. As bile acid it solubilizes cholesterol gallstones and improves the liver function in case of cholestatic diseases. UDCA can be obtained from cholic acid (CA), which is the most abundant and least expensive bile acid available. The now available chemical routes for the obtainment of UDCA yield about 30% of final product. For these syntheses several protection and deprotection steps requiring toxic and dangerous reagents have to be performed, leading to the production of a series of waste products. In many cases the cholic acid itself first needs to be prepared from its taurinated and glycilated derivatives in the bile, thus adding to the complexity and multitude of steps involved of the synthetic process. For these reasons, several studies have been performed towards the development of microbial transformations or chemoenzymatic procedures for the synthesis of UDCA starting from CA or chenodeoxycholic acid (CDCA). This promising approach led several research groups to focus their attention on the development of biotransformations with non-pathogenic, easy-to-manage microorganisms, and their enzymes. In particular, the enzymatic reactions involved are selective hydrolysis, epimerization of the hydroxy functions (by oxidation and subsequent reduction) and the specific hydroxylation and dehydroxylation of suitable positions in the steroid rings. In this minireview, we critically analyze the state of the art of the production of UDCA by several chemical, chemoenzymatic and enzymatic routes reported, highlighting the bottlenecks of each production step. Particular attention is placed on the precursors availability as well as the substrate loading in the process. Potential new routes and recent developments are discussed, in particular on the employment of flow-reactors. The latter technology allows to develop processes with shorter reaction times and lower costs for the chemical and enzymatic reactions involved.

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“…Hydroxylation at C-7 produces very useful hydroxysteroids of pharmacological value (32,33). Although many fungi have been reported to mediate 7-hydroxylation of steroids (34)(35)(36)(37)(38)(39)(40)(41)(42), low stereoselectivity (7␣/7␤) (38,41) and side hydroxylation reactions (37,42) have often been observed, thus presenting great challenges in developing highly regio-and stereoselective 7-hydroxylation processes of steroidal compounds.…”

Section: Discussionmentioning

confidence: 99%

Distinct Regioselectivity of Fungal P450 Enzymes for Steroidal Hydroxylation

Lü

Feng

Chen

et al. 2019

Appl Environ Microbiol

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In this study, we identified two P450 enzymes (CYP5150AP3 and CYP5150AN1) from Thanatephorus cucumeris NBRC 6298 by combination of transcriptome sequencing and heterologous expression in Pichia pastoris. The biotransformation of 11-deoxycortisol and testosterone by Pichia pastoris whole cells coexpressing the cyp5150ap3 and por genes demonstrated that the CYP5150AP3 enzyme possessed steroidal 7β-hydroxylase activities toward these substrates, and the regioselectivity was dependent on the structures of steroidal compounds. CYP5150AN1 catalyzed the 2β-hydroxylation of 11-deoxycortisol. It is interesting that they display different regioselectivity of hydroxylation from that of their isoenzyme, CYP5150AP2, which possesses 19- and 11β-hydroxylase activities. IMPORTANCE The steroidal hydroxylases CYP5150AP3 and CYP5150AN1 together with the previously characterized CYP5150AP2 belong to the CYP5150A family of P450 enzymes with high amino acid sequence identity, but they showed completely different regioselectivities toward 11-deoxycortisol, suggesting the regioselectivity diversity of steroidal hydroxylases of CYP5150 family. They are also distinct from the known bacterial and fungal steroidal hydroxylases in substrate specificity and regioselectivity. Biocatalytic hydroxylation is one of the important transformations for the functionalization of steroid nucleus rings but remains a very challenging task in organic synthesis. These hydroxylases are useful additions to the toolbox of hydroxylase enzymes for the functionalization of steroids at various positions.

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Hydroxylation of lithocholic acid by selected actinobacteria and filamentous fungi

Cited by 25 publications

References 28 publications

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Bacterial steroid hydroxylases: enzyme classes, their functions and comparison of their catalytic mechanisms

Latest development in the synthesis of ursodeoxycholic acid (UDCA): a critical review

Distinct Regioselectivity of Fungal P450 Enzymes for Steroidal Hydroxylation

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