Abstract

Proteomic methods and approaches to the detection of tissue-specific and tissue-generating proteins and peptides – which form corrective properties – in studied meat samples and specially developed meat products were successfully tried out in 2016–2017. The methods allow one to confirm protein and peptide authenticity and also detect bio-markers of proteolytic changes in meat after slaughter. The following proteomic techniques were used in the present research: two-dimensional O’Farrell electrophoresis with isoelectrofocusing in ampholin and immobilin pH gradients, the detection of proteins on two-dimensional electrophoregrams by staining with Coomassie R-250 and silver nitrate, and mass spectrometric identification of proteins by means of MALDI-TOF and MS/MS methods. Contractile actomyosin complex proteins, such as myosin light chains and tropomyosins, were the most informative among proteins of species specificity. It is also necessary to mention that earlier experiments allowed us to choose enzymes which play a part in carbohydrate metabolism (glyceraldehyde 3-phosphate dehydrogenase and β-enolase) as markers. In addition to the listed proteins, myoglobins, actins, and several other proteins in horse meat have showed high species specificity and have been detected well. A system of species specificity (authenticity) of meat raw materials was suggested. The system allows the presence of pork, beef, horse, and camel meat to be detected in both raw and heat-treated products if the content is 5% and more. The data has been obtained by means of bioinformatics, a highly useful tool for formulating an algorithm to identify the protein markers for the Atlas “Proteomic profiles of farm animals meat proteins”. “Proteomic profiles of farm animals muscle proteins”.

Keywords

Beef, pork, horse meat, camel meat, proteomics, muscle proteins, peptide fingerprint, 2D-electrophoresis, MALDI-TOF mass spectrometry, authenticity

REFERENCES

1. Shishkin S.S, Kovalev L.I, Kovaleva M.A., et al. The application of proteomic technologies for the analysis of muscle proteins of farm animals used in the meat industry (Review).Applied Biochemistry and Microbiology, 2014, vol. 50, no. 5, pp. 453–465. (In Russian).
2. Vostrikova N.L. and Chernukha I.M. Bioinformatics – instrument interpretation proteomic profiles of meat protein. Theory and practice of meat processing, 2017, vol. 2, no. 1, pp. 4–17. (In Russian). DOI: 10.21323/2414-438X-2017-2-1-4-17.
3. 3. Chernukha I.M., Fedulova L.V, Kotenkova E.A., Shishkin S.S., and Kovalyov L.I. The Influence of Autolysis on the protein-peptide profile of Bos taurus and Sus scrofa Heart and aorta Tissues. Theory and practice of meat processing, 2016, vol. 1, no. 2, pp. 4–9. (In Russian). DOI:10.21323/2414-438X-2016-1-2-4-9.
4. Picard B., Lebret B., Cassar-Malek I., et al. Recent advances in omic technologies for meat quality amangement. Meat Science, 2015, vol. 109, pp. 18–26. DOI: 10.1016/j.meatsci.2015.05.003.
5. Zhang R., Große-Brinkhaus C., Heidt H., et al. Polymorphisms and expression analysis of SOX-6 in relation to porcine growth, carcass, and meat quality traits. Meat Science, 2015, vol. 109, pp. 18–26. DOI: 10.1016/j.meatsci.2015.04.007.
6. Anderson N.L., Polanski M., Pieper R., et al. The Human Plasma Proteome: A Nonredundant List Developed by Combination of Four Separate Sources. Molecular & Cellular Proteomics, 2004, vol. 3, no. 4, pp. 311–326. DOI: 10.1074/mcp.M300127-MCP200.
7. Shishkin S.S., Kovaleva M.A., Eryomina L.S., Lisitskaya K.V., and Kovalev L.I. Proteomic Approaches for the Study of Transgelins as Tumor-associated Proteins and Potential Biomarkers. Current Proteomics, 2013, vol. 10, no. 2, pp.165–178. DOI: 10.2174/1570164611310020008.
8. Granvogl B., Plöscher M., and Eichacker L.A. Sample preparation by in-gel digestion for mass spectrometry-based proteomics. Analytics and Bioanalytics Chemistry, 2007, vol. 389, no. 4, p. 991–1002. DOI: 10.1007/s00216-007-1451-4.
9. Medzihradszky K.F. In-solution digestion of proteins for mass spectrometry. Methods in Enzymology, 2005, vol. 405, pp. 50–65. DOI: 10.1016/S0076-6879(05)05003-2.
10. Zvereva E.A., Kovalev L.I., Ivanov A.V., et al. Enzyme immunoassay and proteomic characterization of troponin I as a marker of mammalian muscle compounds in raw meat and some meat products. Meat Science, 2015, vol. 105, pp. 46–52. DOI: 10.1016/j.meatsci.2015.03.001.
11. SIB Swiss Institute of Bioinformatics. Available at: http://web.expasy.org/docs/swiss-prot_guideline.html/ UniProtKB/Swiss-Prot/SIB Swiss Institute of Bioinformatics. (accessed 10 May 2017).
12. Anderson N.L. and Anderson N.G. Proteome and proteomics: New technologies, new concepts, and new words. Electrophoresis, 1998, vol. 19, no. 11, pp. 1853–1861. DOI: 10.1002/elps.1150191103.
13. Kovalev L.I., Shishkin S.S., Kovaleva M.A., Vostrikova N.L., and Chernukha I.M. Proteomic research proteins in a sample of pork meat products. All about the meat, 2013, no. 3, pp. 32–34. (In Russian).
14. Manyukhin Ya.S., Chernukha I.M., Kovalev L.I., et al. The study of horsemeat proteins by use proteomic technologies. All about the meat, 2014, no. 3, pp. 20–24. (In Russian).
15. Ivanov A.V. Sravnitel’noe proteomnoe issledovanie belkov cheloveka, uchastvuyush’ikh v obespechenii dvigatel’nykh funktsiy [Comparative proteomic study of human proteins taking part in locomotor functions]. Abstract of Diss. Cand. Sci. (Eng.). Moscow, 2012. 25 p.
16. Manyukhin Ya.S., Chernukha I.M., Vostrikova N.L., et al. The study of muscle proteins of camel using proteomic technologies. All about the meat, 2016, no. 6, pp. 24–28. (In Russian).
17. Melody J.L., Lonergan S.M., Rowe L.J., et al. Early post mortem biochemical factors influence tenderness and water-holding capacity of three porcine muscles. Journal of Animal Science, 2004, vol. 82, no. 4, pp. 1195–1205.
18. Stadtman E.R. Metal ion-catalyzed oxidation of proteins: Biochemical mechanism and biological consequences. Free Radical Biology and Medicine, 1990, vol. 9, no. 4, pp. 315–325. DOI: 10.1016/0891-5849(90)90006-5.
19. Lametsch R. Proteomics in Muscle-to-Meat. Proceedings of the American Meat Science Association 64th Reciprocal Meat Conference. Kansas, 2012, pp. 19–23.
20. Jira W. Aktuelles aus der internationalen Fleischforschung massenspekrtrometrischer Nachweis von Tierarten und Klebefleisch. Fleischwirtschaft, 2014, vol. 94, no. 5, pp. 98–101.
21. Pares D., Saguer E., Pap N., Toldra M., and Carretero C. Low-salt porcine serum concentrate as functional ingredient in frankfurters. Meat Science, 2012, vol. 92, no. 2, pp. 151–156. DOI: 10.1016/j.meatsci.2012.04.029.