Structural characterization of <scp>CYP</scp>260A1 from <i>Sorangium cellulosum</i> to investigate the 1α‐hydroxylation of a mineralocorticoid

Khatri, Yogan; Carius, Yvonne; Ringle, Michael; Lancaster, C. Roy D.; Bernhardt, Rita

doi:10.1002/1873-3468.12479

Cited by 10 publications

(26 citation statements)

References 56 publications

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“…Another product—P3—had a mass of m / z [ M +H] + 361.2007; the calculated mass of molecular formula C 21 H 29 O 5 + was 361.2010. This might be the result of the presence of either a double bond or a keto group or of epoxide formation because both keto groups and epoxides can form from hydroxy groups . It is possible that P3 might be a product of oxidation of P2 at C21 (C−OH→C=O) to give a ketoacid.…”

Section: Resultsmentioning

confidence: 99%

“…Similarly, CYP514C3 from Streptomyces griseus and CYP154C5 from Nocardia francinica can oxidize steroids such as progesterone, testosterone, nandrolone, dehydroepiandrostenedione, and androstenedione at the 16α position . Recently, it has been shown that CYP260A1 and CYP260B1 from Sporangium cellulosum and CYP109E1 from B. megaterium DSM319 can also hydroxylate steroids …”

Section: Introductionmentioning

confidence: 99%

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Tracking Down a New Steroid‐Hydroxylating Promiscuous Cytochrome P450: CYP154C8 from Streptomyces sp. W2233‐SM

et al. 2018

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CYP154C8 from Streptomyces sp. has been identified as a new cytochrome P450 with substrate flexibility towards different sets of steroids. In vitro treatment of these steroids with CYP154C8 revealed interesting product formation patterns with the same group of steroids. NMR study revealed the major product of corticosterone to be hydroxylated at the C21 position, whereas progesterone, androstenedione, testosterone, and 11-ketoprogesterone were exclusively hydroxylated at the 16α position. However, the 16α-hydroxylated product of progesterone was further hydroxylated to yield dihydroxylated products. 16-hydroxyprogesterone was hydroxylated at two positions to yield dihydroxylated products: 2α,16α-dihydroxyprogesterone and 6β,16α-dihydroxyprogesterone. To the best of our knowledge, this is the first report of generation of such products through enzymatic hydroxylation by a CYP450. In view of the importance of modified steroids as pharmaceutical components, CYP154C8 has immense potential for utilization in bioproduction of hydroxylated derivative compounds to be directly employed for pharmaceutical applications.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Tracking Down a New Steroid‐Hydroxylating Promiscuous Cytochrome P450: CYP154C8 from Streptomyces sp. W2233‐SM

et al. 2018

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“…Both the peroxygenases catalyze hydroxylation at the C21 position of cortisone, prednisone, and 11-deoxycortisol substrates and subsequently convert them into possible carboxylic acid that finally leads to C-C bond cleavage forming 17-ketosteroids [14]. It is believed that the possibility of formation of oxidation products could be due to the presence of a double bond or formation of a keto or epoxide, as keto groups and epoxides can form from hydroxyl groups [36,37]. However, in contrast to the pattern of reaction products formation by peroxygenase, with the increase in the time of the reaction catalyzed by CYP154C8, no change in C-C bond cleavage product formation was observed, although sequential hydroxylation of such products (17-ketosteroids) was observed in traces, while increase in the formation of di-hydroxylated product from 21-hydroxyprednisone was observed with the increase in reaction time.…”

Section: Discussionmentioning

confidence: 99%

“…S7). It is believed that the possibility of formation of oxidation products could be due to the presence of a double bond or formation of a keto or epoxide, as keto groups and epoxides can form from hydroxyl groups [36,37].…”

Section: Discussionmentioning

confidence: 99%

Bacterial CYP154C8 catalyzes carbon‐carbon bond cleavage in steroids

Dangi

2018

FEBS Letters

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Here, we report the first bacterial cytochrome P450, CYP154C8, that catalyzes the C–C bond cleavage reaction of steroids. A major change in product distribution is observed with CYP154C8, when the reactions are supported by NADPH and spinach redox partners ferredoxin and ferredoxin reductase, compared with previously reported reactions supported by NADH and redox partners containing putidaredoxin and putidaredoxin reductase. The NMR‐based structural elucidation of reaction products reveals 21‐hydroxyprednisone as the major product for prednisone, while the other product is identified as 1‐dehydroadrenosterone obtained due to C–C bond cleavage. A similar pattern of product formation is observed with cortisone, hydrocortisone, and prednisone. The reaction catalyzed by CYP154C8 in the presence of oxygen surrogates also prominently shows the formation of C–C bond cleavage products.

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“…CYP154C family P450s show 16␣-hydroxylation activities toward various steroidal substrates (13)(14)(15). CYP260A1 of the CYP260A family of P450s is able to convert various C 19 steroids at the 1␣ position, while CYP260B1 has been identified as a 6␤-and 9␣-hydroxylase of C 21 steroids (16,17). On the other hand, although many fungal P450s are referred to as being involved in steroid hydroxylations, not many of them have been biochemically characterized.…”

mentioning

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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Structural characterization of CYP260A1 from Sorangium cellulosum to investigate the 1α‐hydroxylation of a mineralocorticoid

Cited by 10 publications

References 56 publications

Tracking Down a New Steroid‐Hydroxylating Promiscuous Cytochrome P450: CYP154C8 from Streptomyces sp. W2233‐SM

Tracking Down a New Steroid‐Hydroxylating Promiscuous Cytochrome P450: CYP154C8 from Streptomyces sp. W2233‐SM

Bacterial CYP154C8 catalyzes carbon‐carbon bond cleavage in steroids

Distinct Regioselectivity of Fungal P450 Enzymes for Steroidal Hydroxylation

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