Cloning, sequencing, and expression in<i>Escherichia coli</i>of a cytochrome P450 gene from<i>Cunninghamella elegans</i>

Wang, Rongfu; Cao, Wei‐Wen; Khan, Ashraf A.; Cerniglia, Carl E.

doi:10.1111/j.1574-6968.2000.tb09168.x

Cited by 24 publications

(7 citation statements)

References 20 publications

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“…The concept of using microorganisms, and in particular Cunninghamella elegans, as models of mammalian metabolism has been well documented [14][15][16]. It has been proved that Cunninghamella has CYP509A1 enzymes that are synonymous to that involved in xenobiotic detoxification in mammals [17] and can metabolize a wide variety of xenobiotics in a regioand stereo-selective manner similar to mammalian enzyme systems [14,15]. A recent review on C. elegans reports that the fungus shows similarities with mammalian metabolism for a wide variety of drugs, more than a hundred of them [15].…”

Section: Accepted Manuscript Introductionmentioning

confidence: 99%

Biotransformation of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans

Watanabe

Kuzhiumparambil

Winiarski

et al. 2016

Forensic Science International

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Being marketed as "legal" smoking blends or mixtures, synthetic cannabinoids are abused widely owing to its cannabis-like effect. Due to the rapid introduction of new generation analogues of synthetic cannabinoids to escape from legislative/judicial control, the investigation of the metabolic pathways of these substances is of particular importance for drug control, abstinence and forensic toxicology purposes. In this study, the in vitro metabolism of JWH-018, JWH-073 and AM2201 by the fungus Cunninghamella elagans has been investigated with the purpose of validating its potential as a complementary model for investigating synthetic cannabinoid metabolism. JWH-018, JWH-073 and AM2201 were incubated for 72h with C. elegans. Detection of metabolites was based on liquid chromatography-tandem mass spectrometry and high resolution mass spectrometry analysis. C. elegans was found capable of producing the majority of the phase I metabolites observed in earlier in vitro and in vivo mammalian studies as a result of monohydroxylation, dihydroxylation, carboxylation, dehydrogenation, ketone formation, dihydrodiol formation, dihydrodiol formation with N-dealkylation and combinations thereof. C. elegans can thus be a useful and economic model for studying synthetic cannabinoid metabolism.

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Section: Accepted Manuscript Introductionmentioning

confidence: 99%

Biotransformation of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans

Watanabe

Kuzhiumparambil

Winiarski

et al. 2016

Forensic Science International

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“…Cunninghamella elegans (C. elegans) is a fungus species known to metabolise many xenobiotics regio-and stereo-selectively, similar to mammalian metabolism [30]. The fungus has enzymatic activity for both phase I and II enzymes [31] and possesses cytochrome P450 enzymes known as CYP509A1, which are close to CYP51 family [32]. While little is known about the activity of CYP509A1, the fungus is capable of various reactions, such as hydroxylation, carboxylation, dihydrodiol formation, oxidative defluorination, N-dealkylation, glucosidation and sulfation [30,33], including those catalysed by human CYP1A2, CYP2C9, CYP2C19 and CYP2D6 [2,[33][34][35][36][37].…”

Section: Introductionmentioning

confidence: 99%

Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans

Watanabe

Kuzhiumparambil

Nguyen

et al. 2017

AAPS J

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The knowledge of metabolic profile of synthetic cannabinoids is important for the detection of the drugs in urinalysis due to the typical absence or low abundance of parent cannabinoids in human urine. The fungus Cunninghamella elegans has been reported to be a useful tool for metabolism study and thus applicability to synthetic cannabinoids metabolism was examined. In this study, 8-quinolinyl 1-(5-fluoropentyl)-1H-indole-3-carboxylate (5F-PB-22), 8-quinolinyl 1-pentyl-1H-indole-3-carboxylate (PB-22), [1-(5-fluoropentyl)-1H-indol-3-yl](2,2,3,3-tetramethylcyclopropyl)methanone (XLR-11) and (1-pentyl-1H-indol-3-yl)(2,2,3,3-tetramethylcyclopropyl)methanone (UR-144) were incubated with Cunninghamella elegans and the metabolites were identified using liquid chromatographyquadrupole time-of-flight mass spectrometry. The obtained metabolites were compared with reported human metabolites to assess the suitability of the fungus to extrapolate human metabolism. 5F-PB-22 underwent, dihydroxylation, dihydrodiol formation, oxidative defluorination, oxidative defluorination to carboxylic acid, ester hydrolysis and glucosidation, alone and/or in combination. The metabolites of PB-22 were generated by hydroxylation, dihydroxylation, trihydroxylation, dihydrodiol formation, ketone formation, carboxylation, ester hydrolysis and glucosidation, alone and/or in combination. XLR-11 was transformed through hydroxylation, dihydroxylation, aldehyde formation, carboxylation, oxidative defluorination, oxidative defluorination to carboxylic acid and glucosidation, alone and/or in combination. UR-144 was metabolised by hydroxylation, dihydroxylation, trihydroxylation, aldehyde formation, ketone formation, carboxylation, N-dealkylation and combinations. These findings were consistent with previously reported human metabolism except for the small extent of ester hydrolysis observed and absence of glucuronidation. Despite the limitations, Cunninghamella elegans demonstrated the capacity to produce a wide variety of metabolites including some major human metabolites of XLR-11 and UR-144 at high abundance, showing the potential for metabolism of newly emerging synthetic cannabinoids.

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“…31%) to the main mammalian CYP involved in drug detoxification, CYP3A4. In contrast, the previously cloned CYP509A1 12 had a 24% identity to this enzyme. Furthermore, members of the CYP53 family in fungi are associated with 4-hydroxylation of benzoic acid and its derivatives 22 .…”

Section: Resultsmentioning

confidence: 69%

“…One Cunninghamella CYP gene has been cloned and overexpressed in E . coli and the protein confirmed to be a CYP by immunological methods 12 . However, no biochemical assays were conducted and its sequence places it in the family CYP509, members of which are not known to be involved in xenobiotic biotransformation 13 .…”

Section: Introductionmentioning

confidence: 97%

The CYPome of the model xenobiotic-biotransforming fungus Cunninghamella elegans

Palmer-Brown

Miranda-CasoLuengo

Wolfe

et al. 2019

Sci Rep

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The fungus Cunninghamella elegans is recognised as a microbial model of mammalian drug metabolism owing to its ability to catabolise xenobiotic compounds in an analogous fashion to animals. Its ability to produce phase I (oxidative) metabolites of drugs is associated with cytochrome P450 (CYP) activity; however, almost nothing is known about these enzymes in the fungus. In this paper we report the in silico analysis of the genome sequence of C . elegans B9769, which contains 32 genes putatively coding for CYPs. Based on their predicted amino acid sequences these were classified as belonging to CYP509, 5203, 5208, 5313, 5210, 61 and 51 families. Reverse transcription-quantitative PCR revealed that the gene coding for CYP5313D1 was significantly upregulated when C . elegans DSM1908 was cultivated in sabouraud dextrose in contrast to its expression in cells grown in Roswell Park Memorial Institute medium. This corresponded to the fungus’ xenobiotic biotransformation ability when grown in the two media. Heterologous expression of cyp5313D1 in Pichia pastoris resulted in a recombinant strain that biotransformed flurbiprofen to 4′-hydroxyflurbiprofen, the same metabolite generated by C . elegans cultures. This is the first report of a xenobiotic-biotransforming CYP from this biotechnologically important fungus.

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Cloning, sequencing, and expression inEscherichia coliof a cytochrome P450 gene fromCunninghamella elegans

Cited by 24 publications

References 20 publications

Biotransformation of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans

Biotransformation of synthetic cannabinoids JWH-018, JWH-073 and AM2201 by Cunninghamella elegans

Metabolic Profile of Synthetic Cannabinoids 5F-PB-22, PB-22, XLR-11 and UR-144 by Cunninghamella elegans

The CYPome of the model xenobiotic-biotransforming fungus Cunninghamella elegans

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