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The roles of different porcine cytochrome P450 enzymes and cytochrome b5A in skatole metabolism

Published online by Cambridge University Press:  23 November 2011

P. Wiercinska ,
Y. Lou  and
E. J. Squires
P. Wiercinska
Affiliation:
Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1
Y. Lou
Affiliation:
Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1
E. J. Squires*
Affiliation:
Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1
*
†E-mail: [email protected]
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Abstract

Boar taint is the unfavourable odour and taste from pork fat, which results in part from the accumulation of skatole (3-methylindole, 3MI). The key enzymes in skatole metabolism are thought to be cytochrome P450 2E1 (CYP2E1) and cytochrome 2A (CYP2A); however, the cytochrome P450 (CYP450) isoform responsible for the production of the metabolite 6-hydroxy-3-methylindole (6-OH-3MI, 6-hydroxyskatole), which is thought to be involved in the clearance of skatole, has not been established conclusively. The aim of this study was to characterize the role of porcine CYP450s in skatole metabolism by expressing them individually in the human embryonic kidney HEK293-FT cell line. This system eliminates the problems of the lack of specificity of antibodies, inhibitors and substrates for CYP450 isoforms in the pig, and contributions of any other CYP450s that would be present. The results show that pig CYP1A1, CYP2A19, CYP2C33v4, CYP2C49, CYP2E1 and CYP3A and human CYP2E1 (hCYP2E1) are all capable of producing the major skatole metabolite 3-methyloxyindole (3MOI), as well as indole-3-carbinol (I3C), 5-hydroxy-3-methylindole (5-OH-3MI), 6-OH-3MI, 2-aminoacetophenone (2AAP) and 3-hydroxy-3-methyloxindole. CYP2A19 produced the highest amount of the physiologically important metabolite 6-OH-3MI, followed by porcine CYP2E1 and CYP2C49; CYP2A19 also produced more 6-OH-3MI than hCYP2E1. Co-transfection with CYB5A increased the production of skatole metabolites by some of the CYP450s, suggesting that CYB5A plays an important role in the metabolism of skatole. We also show the utility of this expression system to check the specificity of selected substrates and antibodies for porcine CYP450s. Further information regarding the abundance of different CYP450 isoforms is required to fully understand their contribution to skatole metabolism in vivo in the pig.

Keywords

boar taintskatole3-methylindolecytochrome b5Acytochrome P450
Type
Full Paper
Information
animal , Volume 6 , Special Issue 5: Symposium “Sustainable animal production in the tropics” (SAPT), Guadeloupe, France, November 15–18, 2010 , May 2012 , pp. 834 - 845
Copyright
Copyright © The Animal Consortium 2011

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References

Agergaard, N, Laue, A 1992. Absorption from the gastrointestinal tract and liver turnover of skatole. In Measurement and prevention of boar taint in entire male pigs (ed. M Bonneau), pp. 15–16. INRA Editions, Paris.Google Scholar
Anzenbacherová, E, Baranová, J, Zuber, R, Pěchová, A, Anzenbacher, P, Souček, P, Martínková, J 2005. Model systems based on experimental animals for studies on drug metabolism in man: (mini)pig cytochromes P450 3A9 and 2E1. Basic and Clinical Pharmacology and Toxicology 96, 244–245.CrossRefGoogle Scholar
Babol, J, Squires, EJ, Lundström, K 1998. Relationship between oxidation and conjugation metabolism of skatole in pig liver and concentrations of skatole in fat. Journal of Animal Science 76, 829–838.CrossRefGoogle ScholarPubMed
Bæk, C, Hansen-Møller, J, Friis, C, Cornett, C, Hansen, SH 1997. Identification of selected metabolites of skatole in plasma and urine from pigs. Journal of Agricultural Food and Chemistry 45, 2332–2340.CrossRefGoogle Scholar
Balk, MW 1987. Emerging models in the USA: swine, woodchucks and the hairless guinea pig. In Animal models: assessing the scope of their use in biomedical research (ed. J Kawamata and EC Melby), pp. 311–326. Alan R. Liss, New York.Google Scholar
Bertz, RJ, Granneman, GR 1997. Use of in vitro and in vivo data to estimate the likelihood of metabolic pharmacokinetic interactions. Clinical Pharmacokinetics 32, 210–258.CrossRefGoogle ScholarPubMed
Billen, MJ, Squires, EJ 2009. The role of porcine cytochrome b5A and cytochrome b5B in the regulation of cytochrome P45017A1 activities. Journal of Steroid Biochemistry and Molecular Biology 113, 98–104.CrossRefGoogle ScholarPubMed
Bonneau, M 1982. Compounds responsible for boar taint, with special emphasis on androstenone: a review. Livestock Production Science 9, 687–707.CrossRefGoogle Scholar
de Lange, CF, Squires, EJ 1995. Entire males vs castrates for pork production – financial benefits to the producer. Ontario Swine Research Review, 41–44.Google Scholar
Diaz, GJ Squires, EJ 2000a. Metabolism of 3-methylindole by porcine liver microsomes: responsible cytochrome P450 enzymes. Toxicological Sciences 55, 284–292.CrossRefGoogle ScholarPubMed
Diaz, GJ Squires, EJ 2000b. Role of aldehyde oxidase in the hepatic in vitro metabolism of 3-methylindole in pigs. Journal of Agricultural Food and Chemistry 48, 833–837.CrossRefGoogle ScholarPubMed
Diaz, GJ, Skordos, KW, Yost, GS, Squires, EJ 1999. Identification of phase I metabolites of 3-methylindole produced by pig liver microsomes. Drug Metabolism and Disposition 27, 1150–1156.Google ScholarPubMed
Friis, C 1993. Disposition of skatole in male and female pigs. Proceedings of the 5th European ISSX Meeting, Tours, France, 26–29 September, 133p.Google Scholar
Friis, C 1995. Is boar taint related to sex differences or polymorphism of skatole metabolism?. In Proceedings of the EAAP Working Group on Production and Utilization of Meat from Entire Male Pigs. Milton Keynes, UK, September 1995.Google Scholar
Jensen, BB, Jensen, MT 1998. Microbial production of skatole in the digestive tract of entire male pigs. In Skatole and boar taint (ed. WK Jensen), pp. 41–75. Danish Meat Research Institute, Roskilde, Denmark.Google Scholar
Juskevich, JC 1987. Comparative metabolism in food-producing animals: programs sponsored by the center for veterinary medicine. Drug Metabolism Reviews 18, 345–362.CrossRefGoogle Scholar
Kojima, M, Morozumi, T 2004. Cloning of six full-length cDNAs encoding pig cytochrome p450 enzymes and gene expression of these enzymes in the liver and kidney. Journal of Health Science 50, 518–529.CrossRefGoogle Scholar
Madan, A, Usuki, E, Burton, LA, Ogilvie, WB, Parkinson, A 2002. In vitro approaches for studying the inhibition of drug-metabolizing enzymes and identifying the drug-metabolizing enzymes responsible for the metabolism of drugs. In Drugs and the pharmaceutical sciences: drug–drug interactions (ed. AD Rodrigues), pp. 217–294. Marcel Dekker, New York.Google Scholar
Matal, J, Matuskova, Z, Tunkova, A, Anzenbacherová, E, Anzenbacher, P 2009. Porcine CYP2A19, CYP2E1 and CYP1A2 forms are responsible for skatole biotransformation in the reconstituted system. Neuroendocrinology Letters 30 (suppl. 1), 36–40.Google ScholarPubMed
McLaughlin, LA, Ronseaux, S, Finn, RD, Henderson, CL, Wolf, CR 2010. Deletion of microsomal cytochrome b5 profoundly affects hepatic and extrahepatic drug metabolism. Molecular Pharmacology 78, 269–278.CrossRefGoogle ScholarPubMed
Moe, M, Lien, S, Bendixen, C, Hedegaard, J, Hornshøj, H, Berget, I, Meuwissen, HET, Grindflek, E 2008. Gene expression profiles in liver of pigs with extreme high and low levels of androstenone. BMC Veterinary Research 4, 29.CrossRefGoogle ScholarPubMed
Monshouwer, M, van't Klooster, GAE, Nijmeijer, SM, Witkamp, RF, van Miert, ASJPAM 1998. Characterization of cytochrome P450 isoenzymes in primary cultures of pig hepatocytes. Toxicology In Vitro 12, 715–723.CrossRefGoogle ScholarPubMed
Peacock, J, Lou, Y, Lundström, K, Squires, EJ 2008. The effect of a c.-8G>T polymorphism on the expression of cytochrome b5A and boar taint in pigs. Animal Genetics 39, 15–21.CrossRefGoogle ScholarPubMed
Prunier, A, Bonneau, M, von Borell, EH, Cinotti, S, Gunn, M, Fredriksen, B, Giersing, M, Morton, DB, Tuyttens, FAM, Velarde, A 2006. A review of the welfare consequences of surgical castration in piglets and the evaluation of non-surgical methods. Animal Welfare 15, 277–289.CrossRefGoogle Scholar
Rendic, S, Carlo, FJD 1997. Human cytochrome P450 enzymes: a status report summarizing their reactions, substrates, inducers and inhibitors. Drug Metabolism Reviews 29, 413–580.CrossRefGoogle ScholarPubMed
Schenkman, JB, Jansson, I 2003. The many roles of cytochrome b5. Pharmacology and Therapeutics 97, 139–152.CrossRefGoogle ScholarPubMed
Skaanlid, MT, Friis, C 2007. Is bupropion a more specific substrate for porcine CYP2E than chlorzoxazone and p-nitrophenol?. Basic and Clinical Pharmacology and Toxicology 101, 159–162.CrossRefGoogle Scholar
Squires, EJ, Lundström, K 1997. Relationship between cytochrome P450IIEI in liver and levels of skatole and its metabolites in intact male pigs. Journal of Animal Science 75, 2506–2511.CrossRefGoogle Scholar
Terner, MA, Gilmore, WJ, Lou, Y, Squires, EJ 2006. The role of CYP2A and CYP2E1 in the metabolism of 3-methylindole in primary cultured porcine hepatocytes. Drug Metabolism and Disposition 34, 848–854.CrossRefGoogle Scholar
Thornton-Manning, RJ, Ruangyuttikarn, W, Gonzalez, JF, Yost, SG 1991. Metabolic activation of the pneumotoxin, 3-methylindole, by vaccinia-expressed cytochrome P450s. Biochemical and Biophysical Research Communications 181, 100–107.CrossRefGoogle ScholarPubMed
Thornton-Manning, J, Appleton, ML, Gonzalez, FJ, Yost, GS 1996. Metabolism of 3-methylindole by vaccinia-expressed P450 enzymes: correlation of 3-methyleneindolenine formation and protein-binding. Pharmacology and Experimental Therapeutics 276, 21–29.Google ScholarPubMed
Tsiaoussis, J, Newsome, NP, Nelson, JL, Hayes, CP, Plevris, NJ 2001. Which hepatocyte will it be? Hepatocyte choice for bioartificial liver support systems. Liver Transplantation 7, 2–10.CrossRefGoogle ScholarPubMed
Wiercinska, P, Squires, EJ 2010. Chlorzoxazone metabolism by porcine cytochrome P450 enzymes and the effect of cytochrome b5. Drug Metabolism and Disposition 38, 857–862.CrossRefGoogle ScholarPubMed
Yamazaki, H, Nakano, M, Gillam, EMJ, Bell, LC, Guengerich, FP, Shimada, T 1996. Requirements for cytochrome b5 in the oxidation of 7-ethoxycoumarin, CLZ, aniline, and N-nitrosodimethylamine by recombinant cytochrome P450 2E1 and by human liver microsomes. Biochemical Pharmacology 52, 301–309.CrossRefGoogle Scholar
Yamazaki, H, Nakamura, M, Komatsu, T, Ohyama, K, Hatanaka, N, Asahi, S, Shimada, N, Guengerich, FP, Shimada, T, Nakajima, M, Yokoi, T 2002. Roles of NADPH-P450 reductase and apo- and holo-cytochrome b5 on xenobiotic oxidations catalyzed by 12 recombinant human cytochrome P450s expressed in membranes of Escherichia coli. Protein Expression and Purification 24, 329–337.CrossRefGoogle ScholarPubMed