Advertisement

Journal of Industrial Microbiology & Biotechnology

, Volume 44, Issue 10, pp 1471–1481 | Cite as

Evaluation of metabolism of azo dyes and their effects on Staphylococcus aureus metabolome

  • Jinchun SunEmail author
  • Jinshan Jin
  • Richard D. Beger
  • Carl E. Cerniglia
  • Huizhong ChenEmail author
Environmental Microbiology - Original Paper

Abstract

Dyes containing one or more azo linkages are widely applied in cosmetics, tattooing, food and drinks, pharmaceuticals, printing inks, plastics, leather, as well as paper industries. Previously we reported that bacteria living on human skin have the ability to reduce some azo dyes to aromatic amines, which raises potential safety concerns regarding human dermal exposure to azo dyes such as those in tattoo ink and cosmetic colorant formulations. To comprehensively investigate azo dye-induced toxicity by skin bacteria activation, it is very critical to understand the mechanism of metabolism of the azo dyes at the systems biology level. In this study, an LC/MS-based metabolomics approach was employed to globally investigate metabolism of azo dyes by Staphylococcus aureus as well as their effects on the metabolome of the bacterium. Growth of S. aureus in the presence of Sudan III or Orange II was not affected during the incubation period. Metabolomics results showed that Sudan III was metabolized to 4-(phenyldiazenyl) aniline (48%), 1-[(4-aminophenyl) diazenyl]-2-naphthol (4%) and eicosenoic acid Sudan III (0.9%). These findings indicated that the azo bond close to naphthalene group of Sudan III was preferentially cleaved compared with the other azo bond. The metabolite from Orange II was identified as 4-aminobenzene sulfonic acid (35%). A much higher amount of Orange II (~90×) was detected in the cell pellets from the active viable cells compared with those from boiled cells incubated with the same concentration of Orange II. This finding suggests that Orange II was primarily transported into the S. aureus cells for metabolism, instead of the theory that the azo dye metabolism occurs extracellularly. In addition, the metabolomics results showed that Sudan III affected energy pathways of the S. aureus cells, while Orange II had less noticeable effects on the cells. In summary, this study provided novel information regarding azo dye metabolism by the skin bacterium, the effects of azo dyes on the bacterial cells and the important role on the toxicity and/or inactivation of these compounds due to microbial metabolism.

Keywords

Azo dyes Staphylococcus aureus Metabolism Metabolomics 

Notes

Acknowledgements and disclaimer

We thank Drs. Li-Rong Yu and Jing Han for their critical review of this manuscript. This study was funded by National Center for Toxicological Research, United States Food and Drug Administration, and supported in part by appointment (JJ) in the Postgraduate Research Fellowship Program by the Oak Ridge Institute for Science and Education through an interagency agreement between the US Department of Energy and the US Food and Drug Administration. The findings and conclusions in this publication are those of the authors and do not represent FDA positions or policies.

Supplementary material

10295_2017_1970_MOESM1_ESM.doc (188 kb)
Supplementary material 1 (DOC 188 kb)

References

  1. 1.
    Cerniglia CE, Freeman JP, Franklin W, Pack LD (1982) Metabolism of benzidine and benzidine-congener based dyes by human, monkey and rat intestinal bacteria. Biochem Biophys Res Commun 107:1224–1229CrossRefPubMedGoogle Scholar
  2. 2.
    Cerniglia CE, Freeman JP, Franklin W, Pack LD (1982) Metabolism of azo dyes derived from benzidine, 3,3′-dimethyl-benzidine and 3,3′-dimethoxybenzidine to potentially carcinogenic aromatic amines by intestinal bacteria. Carcinogenesis 3:1255–1260CrossRefPubMedGoogle Scholar
  3. 3.
    Cerniglia CE, Zhuo Z, Manning BW, Federle TW, Heflich RH (1986) Mutagenic activation of the benzidine-based dye direct black 38 by human intestinal microflora. Mutat Res 175:11–16CrossRefPubMedGoogle Scholar
  4. 4.
    Chen H, Wang RF, Cerniglia CE (2004) Molecular cloning, overexpression, purification, and characterization of an aerobic FMN-dependent azoreductase from Enterococcus faecalis. Protein Expr Purif 34:302–310CrossRefPubMedGoogle Scholar
  5. 5.
    Chen H, Hopper SL, Cerniglia CE (2005) Biochemical and molecular characterization of an azoreductase from Staphylococcus aureus, a tetrameric NADPH-dependent flavoprotein. Microbiology 151:1433–1441CrossRefPubMedGoogle Scholar
  6. 6.
    Chen YE, Tsao H (2013) The skin microbiome: current perspectives and future challenges. J Am Acad Dermatol 69:143–155CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Childs JJ, Nakajima C, Clayson DB (1967) The metabolism of 1-phenylazo-2-naphthol in the rat with reference to the action of the intestinal flora. Biochem Pharmacol 16:1555–1561CrossRefPubMedGoogle Scholar
  8. 8.
    Chung KT (1983) The significance of azo-reduction in the mutagenesis and carcinogenesis of azo dyes. Mutat Res 114:269–281CrossRefPubMedGoogle Scholar
  9. 9.
    Chung KT, Stevens SE Jr, Cerniglia CE (1992) The reduction of azo dyes by the intestinal microflora. Crit Rev Microbiol 18:175–190CrossRefPubMedGoogle Scholar
  10. 10.
    Cundell AM (2016) Microbial ecology of the human skin. Microb Ecol. doi: 10.1007/s00248-016-0789-6 PubMedGoogle Scholar
  11. 11.
    Dettmer K, Aronov PA, Hammock BD (2007) Mass spectrometry-based metabolomics. Mass Spectrom Rev 26:51–78CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Feng J, Heinze TM, Xu H, Cerniglia CE, Chen H (2010) Evidence for significantly enhancing reduction of Azo dyes in Escherichia coli by expressed cytoplasmic Azoreductase (AzoA) of Enterococcus faecalis. Protein Pept Lett 17:578–584CrossRefPubMedGoogle Scholar
  13. 13.
    Feng J, Cerniglia CE, Chen H (2012) Toxicological significance of azo dye metabolism by human intestinal microbiota. Front Biosci (Elite Ed) 4:568–586CrossRefGoogle Scholar
  14. 14.
    Fouts JR, Kamm JJ, Brodie BB (1957) Enzymatic reduction of prontosil and other azo dyes. J Pharmacol Exp Ther 120:291–300PubMedGoogle Scholar
  15. 15.
    Gingell R, Bridges JW, Williams RT (1969) Gut flora and the metabolism of prontosils in the rat. Biochem J 114:5P–6PCrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Gingell R, Walker R (1971) Mechanisms of azo reduction by Streptococcus faecalis. II. The role of soluble flavins. Xenobiotica 1:231–239CrossRefPubMedGoogle Scholar
  17. 17.
    Griswold DP Jr, Casey AE, Weisburger EK, Weisburger JH (1968) The carcinogenicity of multiple intragastric doses of aromatic and heterocyclic nitro or amino derivatives in young female sprague-dawley rats. Cancer Res 28:924–933PubMedGoogle Scholar
  18. 18.
    Haug W, Schmidt A, Nortemann B, Hempel DC, Stolz A, Knackmuss HJ (1991) Mineralization of the sulfonated azo dye mordant Yellow 3 by a 6-aminonaphthalene-2-sulfonate-degrading bacterial consortium. Appl Environ Microbiol 57:3144–3149PubMedPubMedCentralGoogle Scholar
  19. 19.
    Keck A, Klein J, Kudlich M, Stolz A, Knackmuss HJ, Mattes R (1997) Reduction of azo dyes by redox mediators originating in the naphthalenesulfonic acid degradation pathway of Sphingomonas sp. strain BN6. Appl Environ Microbiol 63:3684–3690PubMedPubMedCentralGoogle Scholar
  20. 20.
    Levine WG (1991) Metabolism of azo dyes: implication for detoxication and activation. Drug Metab Rev 23:253–309CrossRefPubMedGoogle Scholar
  21. 21.
    Li L, Gao HW, Ren JR, Chen L, Li YC, Zhao JF, Zhao HP, Yuan Y (2007) Binding of Sudan II and IV to lecithin liposomes and E. coli membranes: insights into the toxicity of hydrophobic azo dyes. BMC Struct Biol 7:16CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Lowry LK, Tolos WP, Boeniger MF, Nony CR, Bowman MC (1980) Chemical monitoring of urine from workers potentially exposed to benzidine-derived azo dyes. Toxicol Lett 7:29–36CrossRefPubMedGoogle Scholar
  23. 23.
    Mailloux RJ, Beriault R, Lemire J, Singh R, Chenier DR, Hamel RD, Appanna VD (2007) The tricarboxylic acid cycle, an ancient metabolic network with a novel twist. PLoS One 2:e690CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Martin CN, Kennelly JC (1985) Metabolism, mutagenicity, and DNA binding of biphenyl-based azodyes. Drug Metab Rev 16:89–117CrossRefPubMedGoogle Scholar
  25. 25.
    Mueller GC, Miller JA (1949) The reductive cleavage of 4-dimethylaminoazobenzene by rat liver; the intracellular distribution of the enzyme system and its requirement for triphosphopyridine nucleotide. J Biol Chem 180:1125–1136PubMedGoogle Scholar
  26. 26.
    Nony CR, Bowman MC, Cairns T, Lowry LK, Tolos WP (1980) Metabolism studies of an azo dye and pigment in the hamster based on analysis of the urine for potentially carcinogenic aromatic amine metabolites. J Anal Toxicol 4:132–140CrossRefPubMedGoogle Scholar
  27. 27.
    Pan H, Feng J, Cerniglia CE, Chen H (2011) Effects of Orange II and Sudan III azo dyes and their metabolites on Staphylococcus aureus. J Ind Microbiol Biotechnol 38:1729–1738CrossRefPubMedGoogle Scholar
  28. 28.
    Pan H, Xu J, Kweon OG, Zou W, Feng J, He GX, Cerniglia CE, Chen H (2015) Differential gene expression in Staphylococcus aureus exposed to Orange II and Sudan III azo dyes. J Ind Microbiol Biotechnol 42:745–757CrossRefPubMedGoogle Scholar
  29. 29.
    Pearce CI, Christie R, Boothman C, von Canstein H, Guthrie JT, Lloyd JR (2006) Reactive azo dye reduction by Shewanella strain J18 143. Biotechnol Bioeng 95:692–703CrossRefPubMedGoogle Scholar
  30. 30.
    Platzek T, Lang C, Grohmann G, Gi US, Baltes W (1999) Formation of a carcinogenic aromatic amine from an azo dye by human skin bacteria in vitro. Hum Exp Toxicol 18:552–559CrossRefPubMedGoogle Scholar
  31. 31.
    Rinde E, Troll W (1975) Metabolic reduction of benzidine azo dyes to benzidine in the rhesus monkey. J Natl Cancer Inst 55:181–182CrossRefPubMedGoogle Scholar
  32. 32.
    Stingley RL, Zou W, Heinze TM, Chen H, Cerniglia CE (2010) Metabolism of azo dyes by human skin microbiota. J Med Microbiol 59:108–114CrossRefPubMedGoogle Scholar
  33. 33.
    Sun J, Schnackenberg LK, Beger RD (2009) Studies of acetaminophen and metabolites in urine and their correlations with toxicity using metabolomics. Drug Metab Lett 3:130–136CrossRefPubMedGoogle Scholar
  34. 34.
    Sun J, Von Tungeln LS, Hines W, Beger RD (2009) Identification of metabolite profiles of the catechol-O-methyl transferase inhibitor tolcapone in rat urine using LC/MS-based metabonomics analysis. J Chromatogr B Anal Technol Biomed Life Sci 877:2557–2565CrossRefGoogle Scholar
  35. 35.
    Sun J, Von Tungeln LS, Hines W, Beger RD (2009) Identification of metabolite profiles of the catechol-O-methyl transferase inhibitor tolcapone in rat urine using LC/MS-based metabonomics analysis. J Chromatogr B Analyt Technol Biomed Life Sci 877:2557–2565CrossRefPubMedGoogle Scholar
  36. 36.
    Sun J, Schnackenberg LK, Hansen DK, Beger RD (2010) Study of valproic acid-induced endogenous and exogenous metabolite alterations using LC–MS-based metabolomics. Bioanalysis 2:207–216CrossRefPubMedGoogle Scholar
  37. 37.
    Sun J, Jin J, Beger RD, Cerniglia CE, Yang M, Chen H (2016) Metabolomics evaluation of the impact of smokeless tobacco exposure on the oral bacterium Capnocytophaga sputigena. Toxicol Vitro 36:133–141CrossRefGoogle Scholar
  38. 38.
    Walker R (1970) The metabolism of azo compounds: a review of the literature. Food Cosmet Toxicol 8:659–676CrossRefPubMedGoogle Scholar
  39. 39.
    Wilson ID, Nicholson JK (2017) Gut microbiome interactions with drug metabolism, efficacy, and toxicity. Transl Res 179:204–222CrossRefPubMedGoogle Scholar
  40. 40.
    Xu H, Heinze TM, Chen S, Cerniglia CE, Chen H (2007) Anaerobic metabolism of 1-amino-2-naphthol-based azo dyes (Sudan dyes) by human intestinal microflora. Appl Environ Microbiol 73:7759–7762CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Xu H, Heinze TM, Paine DD, Cerniglia CE, Chen H (2010) Sudan azo dyes and Para Red degradation by prevalent bacteria of the human gastrointestinal tract. Anaerobe 16:114–119CrossRefPubMedGoogle Scholar
  42. 42.
    Yoshida O (1973) Etiological factors in bladder tumors. Nihon Hinyokika Gakkai Zasshi 64:707–712PubMedGoogle Scholar
  43. 43.
    Zimmermann T, Kulla HG, Leisinger T (1982) Properties of purified Orange II azoreductase, the enzyme initiating azo dye degradation by Pseudomonas KF46. Eur J Biochem 129:197–203CrossRefPubMedGoogle Scholar
  44. 44.
    Zimmermann T, Gasser F, Kulla HG, Leisinger T (1984) Comparison of two bacterial azoreductases acquired during adaptation to growth on azo dyes. Arch Microbiol 138:37–43CrossRefPubMedGoogle Scholar

Copyright information

© © Society for Industrial Microbiology and Biotechnology (outside the USA) 2017

Authors and Affiliations

  1. 1.Division of Systems BiologyNational Center for Toxicological Research, US FDAJeffersonUSA
  2. 2.Division of MicrobiologyNational Center for Toxicological Research, US FDAJeffersonUSA

Personalised recommendations