The cytotoxic and genotoxic effects of conjugated trans-2-nonenal [T2N], an off-flavor compound in beer and-heat processed food arising from lipid oxidation

• Autorzy: Szwajcer Dey E , Staniszewska M. , Pasciak M. , Konopacka M. , Rogolinski J. , Gamian A. , Danielsson B.
• Języki publikacji: EN

Abstrakty

EN

This study investigates the toxic effect of E(2)nonenal (trans-2-nonenal, T2N) and its conjugate with horse muscle myoglobin (Mb) tested on murine cell line L₉₂₉ and human cell line A₅₄₉, as well as the genotoxic effect of these compounds assayed by measuring of micronuclei in human cells K₅₆₂. It is an aldehyde, which is occurring as the substance responsible for an off flavour in aged beers, but originates also from lipid oxidation in heat processed food. T2N is an aldehyde formed from linoleic acid as a secondary oxidation product. The modification of Mb with T2N was analyzed with the use of SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and electrospray ionization mass spectrometry (ESI-MS). Results from SDS-PAGE suggest that T2N substitutes Mb and additionally causes cross-linking with polymerization of Mb resulting in an insoluble fraction. The ESI-MS spectrum of the soluble fraction used in the toxicity tests, demonstrated that conjugation of T2N with Mb yielded Mb adducts with one residue of trans-2-nonenal per myoglobin molecule as the major fraction and adducts with different numbers of T2N molecules as minor fractions. In the cytotoxicity assay the T2N and its Mb conjugate causes 50 % destruction of cells at the concentration 95-125 pg/ml and 200 pg/ml respectively, when L₉₂₉ and A₅₄₉ cell lines were used, whereas Mb control tested up to 2000 mg/ml was without any cytotoxic effect. In genotoxicity in vitro assay we have observed that the T2N and its Mb conjugate expressed the genotoxicity. The number of micronuclei in human K₅₆₂ cells reac₅₆₂ 26 ± 2.16 promille (MN/1000 cells), comparing to 62 ± 8.64 MN/1000 cells for the reference free T2N, whereas a control value was 10.33 ± 1.25 MN/1000 cells. The studied compounds expressed also the apoptotic effect in K₅₆₂ cells as the number of apoptotic cells increased to 44.67 ±4.92 promille for T2N-Mb, comparing to 168.67 ±37.28 promille for free T2N, whereas a control value was 30.33 ± 1.36 promille for Mb. In these assays the T2N-Mb conjugate is several times more toxic in relation to control protein. Results indicate that T2N adducts with protein are potent to induce various cytotoxic and apoptotic effects when assayed in vitro tests. It suggests that higher level of such aldehyde might create in organism severe potential of toxicity.

Słowa kluczowe

EN

horse muscle; genotoxicity; heat processed food; lipid oxidation; myoglobin; toxic effect; trans-2-nonenal; apoptosis; food chemistry; cytotoxicity

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2005
• Tom: 54
• Numer: Suppl.
• Opis fizyczny: p.47-52,fig.,ref.

Twórcy

autor

Szwajcer Dey E

• Lund University, Box 124, 221 00 Lund, Sweden

autor

Staniszewska M.

autor

Pasciak M.

autor

Konopacka M.

autor

Rogolinski J.

autor

Gamian A.

autor

Danielsson B.

Bibliografia

• Abend M., A. Rhein, K.P. Gilbertz, W.F. Blakely and D. Van Beuningen. 1995. Correlation of micronucleus and apoptosis assays with reproductive cell death. Int. J. Radiat. Biol. 67: 315-326.
• Back W., C. Forster, M. Krollenthaler, J. Lehmann, B. Sacher, B. Thum and F. Weihenstephan. 1999. New research findings on improving taste stability. Brauwelt International pp. 380-390.
• Drost B.W., R. Van den Berg, F.J.M. Freijee, E.G. Van der Velde and M.J. Hollemans. 1990. Flavour stability.Soc. Brew. Chem. 48: 124-131.
• Esterbauer FL, R.J. Schaur and H. Zollner. 1991. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Review Article Free Radio. Biol. Med. 11: 81-128.
• Faustman C, D.C. Liebler, D. McClure and Q. Sun. 1999. α,ß-unsaturated aldehydes accelerate oxymyoglobin oxidation. J. Agric. Food Chem. 47: 3140-3144.
• Guillen-Sans R. and M. Guzman-Chozas. 1998. The thiobarbituric Acid (TBA) reaction in foods: A review. Crit. Rev. Food Sci. Nutrition 38: 315-330.
• Hilderbrand D.F. 1992. Altering fatty acid metabolism in plants. Food Technol. 71-74.
• Laemmli U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T. Nature 227: 680-685.
• Lermusieau G., S. Noel and C. Liegeois. 1999. Nonoxidative mechanism for development of trans-2-nonenal in beer. J. Am. Soc. Brew. Chem. 57: 29-33.
• Lynch M.P. and C. Faustman. 2000. Effect of aldehyde lipid oxidation products on myoglobin. J. Agric. Food Chem. 48: 600-604.
• Noel S., C. Liegeois, G. Lermusieau, E. Bodart, C. Badot and S. Collin. 1999. Release of deuterated nonenal during beer aging from labelled precursors synthesized in the boiling kettle. J. Agric. Food Chem. 47: 4323-4326.
• Nyborg M., H. Outtrup and T. Dreyen 1999. Investigations of the protective mechanism of sulfite against beer staling and formation of adducts with trans-2-nonenal. J. Am. Soc. Brew. Chem. 57: 24-28.
• Piasecki E., A.D. Inglot, J. Zielińska-Jenczylik, J. Młochowski and L. Syper. 1992. Simultaneous induction of interferon gamma and tumour necrosis factor alpha by different seleno-organic compounds in human peripheral blood leukocytes. Arch. Immunol. Then Exp. 40: 229-234.
• Sakai T., S. Kuwazuru, K. Yamauchi and K.A. Uchida. 1995. Lipid peroxidation-derived aldehyde, 4-hydroxy-2-nonenal and W-6 fatty acids contents in meat. Biosci. Biotechnol. Biochem. 59: 1379-1380.
• Tressl R., D. Bahri and R. Silwan 1979. Bildung von Aldehyden durch Lipoxydation under deren Bedeutung als "Off-flavour" komponenten in Bier. Proceedings of the European Brewery Convention Congress 17: 27-41.