European Food Research and Technology

, Volume 234, Issue 4, pp 663–670 | Cite as

Pro-oxidative effects of melanoidin–copper complexes on isolated and cellular DNA

  • Bettina Cämmerer
  • Katharina Chodakowski
  • Claudia Gienapp
  • Laura Wohak
  • Andrea Hartwig
  • Lothar W. Kroh
Original paper


High molecular weight Maillard reaction products (melanoidins) are described to possess metal-chelating properties. Whereas in food systems, this ability contributes to antioxidant properties, the consequences on biological systems are not quite clear. The study was aimed to evaluate the implication of metal complexation by melanoidins on DNA damage. Melanoidins prepared with d-glucose and different l-amino acids under water-free reaction conditions were charged with cupric ions. The effect on isolated DNA was investigated by the PM2 assay and on cellular systems in the human colon carcinoma cell line HCT-116 by alkaline unwinding. Independent of the amino acid composition, pure melanoidins (MW >14 kDa) did not cause significant DNA damage. By charging melanoidins with Cu2+ ions, a considerable DNA strand breaking activity was detectable, which was again amplified in an oxidative milieu (addition of hydrogen peroxide). Since Cu2+ normally does not provoke the formation of reactive oxygen species (ROS) via Fenton-type reaction, the results obtained have to be attributed to reducing properties of melanoidins. Thus, in melanoidin–copper complexes redox cycling may take place leading to Cu+ which subsequently undergoes Fenton-type and Haber–Weiss reactions. As a consequence, ROS are formed, which may explain the generation of DNA strand breaks.


Melanoidins Metal chelating DNA strand breaks Fenton reaction PM2-assay Pro-oxidative action 


  1. 1.
    Gomyo T, Horikoshi M (1976) On the interaction of melanoidin with metallic ions. Agr Biol Chem 40:33–40CrossRefGoogle Scholar
  2. 2.
    Rendleman JA Jr (1987) Complexation of calcium by melanoidin and its role in determining bioavailability. J Food Sci 52:1699–1705CrossRefGoogle Scholar
  3. 3.
    Morales FJ, Fernandez-Fraguas C, Jimenez-Perez S (2005) Iron-binding ability of melanoidins from food and model systems. Food Chem 90:821–827CrossRefGoogle Scholar
  4. 4.
    Wijewickreme AN, Kitts DD, Durance TD (1997) Reaction conditions influence the elementary composition and metal chelating affinity of nondialyzable model Maillard reaction products. J Agric Food Chem 45:4577–4583CrossRefGoogle Scholar
  5. 5.
    O’Brien J, Morrissey PA (1997) Metal ion complexation by products of the Maillard reaction. Food Chem 58:17–27CrossRefGoogle Scholar
  6. 6.
    Rendleman JA, Inglett GE (1990) The influence of Cu2+ in the Maillard reaction. Carbohydr Res 201:311–326CrossRefGoogle Scholar
  7. 7.
    Cosovic B, Vojvodic V, Boskovic N, Plavsic M, Lee C (2010) Characterization of natural and synthetic humic substances (melanoidins) by chemical composition and adsorption measurements. Org Geochem 41:200–205CrossRefGoogle Scholar
  8. 8.
    Plavsic M, Cosovic B, Lee C (2006) Copper complexing properties of melanoidins and marine humic material. Sci Total Environ 366:310–319CrossRefGoogle Scholar
  9. 9.
    Yoshimura Y, Iijima T, Watanabe T, Nakazawa H (1997) Antioxidative effect of Maillard reaction products using glucose-glycine model system. J Agric Food Chem 45:4106–4109CrossRefGoogle Scholar
  10. 10.
    Borrelli RC, Fogliano V, Monti SM, Ames JM (2002) Characterization of melanoidins from a glucose-glycine model system. Eur Food Res Technol 215:210–215CrossRefGoogle Scholar
  11. 11.
    Delgado-Andrade C, Seiquer I, Navarro MP (2004) Bioavailability of iron from a heat treated glucose-lysine model food system: assays in rats and in Caco-2 cells. J Sci Food Agric 84:1507–1513CrossRefGoogle Scholar
  12. 12.
    Seiquer I, Aspe T, Vaquero P, Navarro MP (2001) Effects of heat treatment of casein in the presence of reducing sugars on calcium bioavailability: in vitro and in vivo assays. J Agric Food Chem 49:1049–1055CrossRefGoogle Scholar
  13. 13.
    Garcia MM, Seiquer I, Delgado-Andrade C, Galdo G, Navarro MP (2009) Intake of Maillard reaction products reduces iron bioavailability in male adolescents. Mol Nutr Food Res 53:1551–1560CrossRefGoogle Scholar
  14. 14.
    Homma S, Murata M (1995) Characterization of metal-chelating compounds in instant coffee. In: ASIC, 19th Colloque, Kyoto, pp 183–191Google Scholar
  15. 15.
    Navarro PM, Aspe T, Seiquer I (2000) Zinc transport in Caco-2 cells and zinc balance in rats: influence of the heat treatment of a casein-glucose-fructose mixture. J Agric Food Chem 48:3589–3596CrossRefGoogle Scholar
  16. 16.
    Wijewickreme AN, Kitts DD (1997) Influence of reaction conditions on the oxidative behavior of model Maillard reaction products. J Agric Food Chem 45:4571–4576CrossRefGoogle Scholar
  17. 17.
    Rivero D, Perez-Magarino S, Gonzales-Sanjose ML, Valls-Belles V, Codoner P, Muniz P (2005) Inhibition of induced DNA oxidative damages by beers: correlation with the content of polyphenols and melanoidins. J Agric Food Chem 53:3637–3642CrossRefGoogle Scholar
  18. 18.
    Rehner G, Walter T (1991) Wirkung von Maillard Produkten und Lysinoalanin auf die Bioverfügbarkeit von Eisen, Kupfer und Zink. Z Ernährungswiss 30:50–55CrossRefGoogle Scholar
  19. 19.
    Maillard M-N, Billaud C, Chow Y-N, Ordonaud C, Nicolas J (2007) Free radical scavenging, inhibition of polyphenoloxidase activity and copper chelating properties of model Maillard systems. LWT Food Sci Technol 40:1434–1444CrossRefGoogle Scholar
  20. 20.
    Rufián-Henares JA, de la Cueva SP (2009) Antimicrobial activity of coffee melanoidins-a study of their metal-chelating properties. J Agric Food Chem 57:432–438CrossRefGoogle Scholar
  21. 21.
    Einarsson H, Snygg BG, Eriksson C (1983) Inhibition of bacterial growth by Maillard reaction products. J Agric Food Chem 31:1043–1047CrossRefGoogle Scholar
  22. 22.
    Rufian-Henares JA, Morales FJ (2008) Antimicrobial activity of melanoidins against Escherichia coli is mediated by a membrane-damage mechanism. J Agric Food Chem 56:2357–2362CrossRefGoogle Scholar
  23. 23.
    Cämmerer B, Kroh LW (1995) Investigation of the influence of reaction conditions on the elementary composition of melanoidins. Food Chem 53:55–59CrossRefGoogle Scholar
  24. 24.
    Schwerdtle T, Walter I, Mackiw I, Hartwig A (2003) Induction of oxidative DNA damage by arsenite and its trivalent and pentavalent methylated metabolites in cultured human cells and isolated DNA. Carcinogenesis 24:967–974CrossRefGoogle Scholar
  25. 25.
    Hartwig A, Dally H, Schlepegrell R (1996) Sensitive analysis of oxidative DNA damage in mammalian cells: use of the bacterial Fpg protein in combination with alkaline unwinding. Toxicol Lett 88:85–90CrossRefGoogle Scholar
  26. 26.
    Rohn S, Rawel HM, Kroll J (2004) Antioxidant activity of protein-bound quercetin. J Agric Food Chem 52:4725–4729CrossRefGoogle Scholar
  27. 27.
    Cämmerer B, Kroh LW (1996) Investigation of the contribution of radicals to the mechanism of the early stage of the Maillard reaction. Food Chem 57:217–221CrossRefGoogle Scholar
  28. 28.
    Yen G-C, Liao C-M, Wu S-C (2002) Influence of Maillard reaction products on DNA damage in human lymphocytes. J Agric Food Chem 50:2970–2976CrossRefGoogle Scholar
  29. 29.
    Taylor JLS et al (2004) Gentoxicity of melanoidin fractions derived from a standard glucose/glycine model. J Agric Food Chem 52:318–323CrossRefGoogle Scholar
  30. 30.
    Wagner K-H, Reichhold S, Koschutnig K, Chériot S, Billaud C (2007) The potential antimutagenic and antioxidant effects of Maillard reaction products used as “natural antibrowning” agents. Mol Nutr Food Res 51:496–504CrossRefGoogle Scholar
  31. 31.
    Glösl S et al (2004) Genotoxicity and mutagenicity of melanoidins isolated from a roasted glucose-glycine model in human lymphocyte cultures, intestinal Caco-2 cells and in the Salmonella typhimurium strains TA98 and TA102 applying the AMES test. Food Chem Toxicol 42:1487–1495CrossRefGoogle Scholar
  32. 32.
    Cheriot S, Billaud C, Pöchtrager S, Wagner K-H, Nicolas J (2009) A comparison study between antioxidant and mutagenic properties of cysteine glucose-derived Maillard reaction products and neoformed products from heated cysteine and hydroxymethylfurfural. Food Chem 114:132–138CrossRefGoogle Scholar
  33. 33.
    Yen G-C, Tsai L-C, Lii J-D (1992) Antimutagenic effect of Maillard Browning products obtained from amino acids and sugars. Food Chem Toxicol 30:127–132Google Scholar
  34. 34.
    Summa C et al (2008) Radical scavenging activity, anti-bacterial and mutagenic effects of cocoa bean Maillard reaction products with degree of roasting. Mol Nutr Food Res 52:342–351CrossRefGoogle Scholar
  35. 35.
    Manzocco L, Calligaris S, Mastrocola D, Nicoli MC, Lerici CR (2001) Review of nonenzymatic browning and antioxidant capacity in processed foods. Trends Food Sci Technol 11:340–346CrossRefGoogle Scholar
  36. 36.
    Rufián-Hernares JA, Delgado-Andrade C, Moralez FJ (2006) Assessing the antioxidant and pro-oxidant activity of phenolic compounds by means of their copper reducing activity. Eur Food Res Technol 223:225–231CrossRefGoogle Scholar
  37. 37.
    Gunther MR, Hanna PM, Mason RP, Cohen MS (1995) Hydroxyl radical formation from cuprous ion and hydrogen peroxide: a spin-trapping study. Arch Biochem Biophys 316:515–522CrossRefGoogle Scholar
  38. 38.
    Kennedy LJ, Moore K, Caulfield JL, Tannenbaum SR, Dedon PC (1997) Quantitation of 8-oxoguanine and strand breaks produced by four oxidizing agents. Chem Res Toxicol 10:386–392CrossRefGoogle Scholar
  39. 39.
    Schwerdtle T et al (2007) Impact of copper on the induction and repair of oxidative DNA damage, poly(ADP-ribosyl)ation and PARP-1 activity. Mol Nutr Food Res 51:201–210CrossRefGoogle Scholar
  40. 40.
    Hamann I, Schwerdtle T, Hartwig A (2009) Establishment of a non-radioactive cleavage assay to assess the DNA repair capacity towards oxidatively damaged DNA in subcellular and cellular systems and the impact of copper. Mutat Res 669:122–130CrossRefGoogle Scholar
  41. 41.
    Macomber L, Rensing C, Imlay JA (2007) Intracellular copper does not catalyze the formation of oxidative DNA damage in Escherichia coli. J Bacteriol 189:1616–1626CrossRefGoogle Scholar
  42. 42.
    Gorren ACF, Schrammel A, Schmidt K, Mayer B (1996) Decomposition of S-nitrosoglutathione in the presence of copper ions and glutathione. Arch Biochem Biophys 330:219–228CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Bettina Cämmerer
    • 1
  • Katharina Chodakowski
    • 1
  • Claudia Gienapp
    • 1
  • Laura Wohak
    • 1
  • Andrea Hartwig
    • 2
  • Lothar W. Kroh
    • 1
  1. 1.Institute of Food Technology and Food ChemistryBerlin University of TechnologyBerlinGermany
  2. 2.Institute of Applied BiosciencesKarlsruhe University of Technology (KIT)KarlsruheGermany

Personalised recommendations