Identification and characterization of a bacterial cytochrome P450 for the metabolism of diclofenac

Prior, Jamie E.; Shokati, Touraj; Christians, Uwe; Gill, Ryan T.

doi:10.1007/s00253-009-2135-0

Cited by 34 publications

(28 citation statements)

References 60 publications

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“…The CYP encoded by sare1259 is most closely related to rif-orf13 and rif-orf5 from the AM- rif cluster as well as other CYPs belonging to the CYP107 family. This superfamily includes a large number of bacterial CYPs, many of which are associated with xenobiotic degradation and secondary metabolite tailoring 29-32…”

Section: Discussionmentioning

confidence: 99%

Shared Biosynthesis of the Saliniketals and Rifamycins in Salinispora arenicola is Controlled by the sare1259-Encoded Cytochrome P450

et al. 2010

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Saliniketals A and B are unusual polyketides from the marine actinomycete Salinispora arenicola that inhibit ornithine decarboxylase induction. The structural similarities between the saliniketals and the ansa chain of the potent rifamycin antibiotics, which co-occur in the fermentation broth, suggest a common origin between the two compound classes. Using PCR-directed mutagenesis, chemical complementation studies, and stable isotope feeding experiments, we showed that the saliniketals are byproducts of the rifamycin biosynthetic pathway diverging at the stage of 34a-deoxyrifamycin W. Our results suggest that a single enzyme, the cytochrome P450 monooxygenase encoded by sare1259, catalyzes multiple oxidative rearrangement reactions on 34a-deoxyrifamyin W to yield both the saliniketal and rifamycin structural classes.

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Section: Discussionmentioning

confidence: 99%

Shared Biosynthesis of the Saliniketals and Rifamycins in Salinispora arenicola is Controlled by the sare1259-Encoded Cytochrome P450

et al. 2010

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“…Studies with native enzymes originating from specialized or pure cultures help to reliably assign function to unknown proteins, thereby improving database annotations (Mills et al., 2015) and mechanistic understanding of degradation processes (Prior et al., 2009, Xu et al., 2015). However, pure or enriched laboratory cultures cannot provide reliable information about the diversity and activity of the enzymes actually involved in micropollutant degradation under environmental conditions.…”

Section: Introductionmentioning

confidence: 99%

Micropollutant degradation via extracted native enzymes from activated sludge

et al. 2016

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A procedure was developed to assess the biodegradation of micropollutants in cell-free lysates produced from activated sludge of a municipal wastewater treatment plant (WWTP). This proof-of-principle provides the basis for further investigations of micropollutant biodegradation via native enzymes in a solution of reduced complexity, facilitating downstream protein analysis. Differently produced lysates, containing a variety of native enzymes, showed significant enzymatic activities of acid phosphatase, β-galactosidase and β-glucuronidase in conventional colorimetric enzyme assays, whereas heat-deactivated controls did not. To determine the enzymatic activity towards micropollutants, 20 compounds were spiked to the cell-free lysates under aerobic conditions and were monitored via LC-ESI-MS/MS. The micropollutants were selected to span a wide range of different biodegradabilities in conventional activated sludge treatment via distinct primary degradation reactions. Of the 20 spiked micropollutants, 18 could be degraded by intact sludge under assay conditions, while six showed reproducible degradation in the lysates compared to the heat-deactivated negative controls: acetaminophen, N-acetyl-sulfamethoxazole (acetyl-SMX), atenolol, bezafibrate, erythromycin and 10,11-dihydro-10-hydroxycarbamazepine (10-OH-CBZ). The primary biotransformation of the first four compounds can be attributed to amide hydrolysis. However, the observed biotransformations in the lysates were differently influenced by experimental parameters such as sludge pre-treatment and the addition of ammonium sulfate or peptidase inhibitors, suggesting that different hydrolase enzymes were involved in the primary degradation, among them possibly peptidases. Furthermore, the transformation of 10-OH-CBZ to 9-CA-ADIN was caused by a biologically-mediated oxidation, which indicates that in addition to hydrolases further enzyme classes (probably oxidoreductases) are present in the native lysates. Although the full variety of indigenous enzymatic activity of the activated sludge source material could not be restored, experimental modifications, e.g. different lysate filtration, significantly enhanced specific enzyme activities (e.g. >96% removal of the antibiotic erythromycin). Therefore, the approach presented in this study provides the experimental basis for a further elucidation of the enzymatic processes underlying wastewater treatment on the level of native proteins.

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“…2; Supplemental Table 2). These bacterial P450 enzymes are considered to be active drug metabolizers for several drugs including amitriptyline, chlorpromazine (2), and diclofenac (5) (Prior et al, 2010;Kulig et al, 2015). In addition, we have previously shown that CYP264A1 was able to convert tricyclic drug molecules (Litzenburger et al, 2015), and CYP265A1 and CYP266A1 were able to hydroxylate the antitumor drug epothilone D .…”

Section: Bioinformatics Studies and Comparison Of Cyp267a1 With Cyp267b1mentioning

confidence: 99%

CYP267A1 and CYP267B1 from Sorangium cellulosum So ce56 are Highly Versatile Drug Metabolizers

Kern

Khatri

Litzenburger

et al. 2016

Drug Metabolism and Disposition

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The guidelines of the Food and Drug Administration and International Conference on Harmonization have highlighted the importance of drug metabolites in clinical trials. As a result, an authentic source for their production is of great interest, both for their potential application as analytical standards and for required toxicological testing. Since we have previously shown promising biotechnological potential of cytochromes P450 from the soil bacterium Sorangium cellulosum So ce56, herein we investigated the CYP267 family and its application for the conversion of commercially available drugs including nonsteroidal anti-inflammatory, antitumor, and antihypotensive drugs. The CYP267 family, especially CYP267B1, revealed the interesting ability to convert a broad range of substrates. We established substrate-dependent extraction protocols and also optimized the reaction conditions for the in vitro experiments and Escherichia coli-based whole-cell bioconversions. We were able to detect activity of CYP267A1 toward seven out of 22 drugs and the ability of CYP267B1 to convert 14 out of 22 drugs. Moderate to high conversions (up to 85% yield) were observed in our established whole-cell system using CYP267B1 and expressing the autologous redox partners, ferredoxin 8 and ferredoxin-NADP + reductase B. With our existing setup, we present a system capable of producing reasonable quantities of the human drug metabolites 49-hydroxydiclofenac, 2-hydroxyibuprofen, and omeprazole sulfone. Due to the great potential of converting a broad range of substrates, wild-type CYP267B1 offers a wide scope for the screening of further substrates, which will draw further attention to future biotechnological usage of CYP267B1 from S. cellulosum So ce56.

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Identification and characterization of a bacterial cytochrome P450 for the metabolism of diclofenac

Cited by 34 publications

References 60 publications

Shared Biosynthesis of the Saliniketals and Rifamycins in Salinispora arenicola is Controlled by the sare1259-Encoded Cytochrome P450

Shared Biosynthesis of the Saliniketals and Rifamycins in Salinispora arenicola is Controlled by the sare1259-Encoded Cytochrome P450

Micropollutant degradation via extracted native enzymes from activated sludge

CYP267A1 and CYP267B1 from Sorangium cellulosum So ce56 are Highly Versatile Drug Metabolizers

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