FARKLI TİPTE UN VE EKMEK ÖRNEKLERİNDE GLUTEN YAPISININ PROTEOMİK VE FT-IR SPEKTROSKOPİSİ ARAÇLARI İLE ARAŞTIRILMASI

Öz

Bu çalışma kapsamında; gluten proteininin alt birimi olan gliadin 2D-PAGE ve FT-IR tekniklerini içeren bazı proteomik yöntemler aracılığı ile araştırılmış ve bu proteinin α ve β yaprak formları arasında dönüşüm olduğu tespit edilmiştir. Denemelerde yer alan örneklerin protein konsantrasyonları un çeşidine ve örneklerin pişmiş formuna göre değişiklik göstermiştir. FT-IR ile elde edilen spektrumlarda Amid A, Amid I ve fosforile protein bölgeleri olmak üzere üç alan çalışılmıştır. Bu spektrum bilgisinden yola çıkarak, gliadin yapısının, ham maddenin ürüne dönüşme sürecinde önemli ölçüde farklılaştığı gözlenmiştir. Beyaz unun ekmek formunda N-H bağı ile ilgili Amid A titreşiminde artış gözlenirken, un örnekleri pişmiş forma dönüştüğünde C=O bağı titreşimi ile bağlantılı olan Amid I bölgesinde azalma tespit edilmiştir. Çalışma sonuçlarından yola çıkarak, ekmek üretiminde kullanılan un çeşidinin ve pişirme şeklinin nihai ürünün gluten yapısı ve miktarı üzerinde etkili olduğu söylenebilir.

Anahtar Kelimeler

• GLUTEN
• GLİADİN
• PROTEOMİKS
• 2D-PAGE
• FT-IR

PROTEOMIC VIEW ON GLUTEN STRUCTURE IN DIFFERENT TYPES OF FLOUR AND BREAD SAMPLES BY USING BOTTOM UP PROTEOMICS AND FT-IR SPECTROSCOPY

Öz

In the current study, some proteomic methods containing 2D-PAGE and FT-IR techniques were performed to screen gliadin, the subunit of gluten protein, and transformation between α and β sheet forms of this protein was evaluated. The protein concentration of the samples varied between flour types and also cooked form of these samples. We focused on Amide A, Amide I and phosphorylated protein regions on the spectrums achieved by FT-IR. Gliadin structure was dramatically differed when the raw material was formed in baked form. While Amide A vibration which is related to N-H streching increased for the bread form of the white flour, Amide I which is related to C=O streching decreased when the raw material changed in the cooked form. It can be concluded that the type of flour used in bread production and the type of baking were effective on gluten structure and amount of the final product.

Anahtar Kelimeler

• GLUTEN
• GLIADIN
• PROTEOMICS
• 2D-PAGE
• FT-IR

Kaynakça

1. Artık, N. (1988). Buğday protein fraksiyonlarının jel elektroforez (SDS-PAGE) ile analizi, aminoasit bileşim ve elektron mikroskobik görünüşlerinin belirlenmesi. Gıda 13(1), 65-75.
2. Bascuñán, K.A., Araya, M., Roncoroni, L., Doneda, L., Elli, L. (2020). Dietary Gluten as a Conditioning Factor of the Gut Microbiota in Celiac Disease. Adv Nutr. 11(1):160-174. doi:10.1093/advances/nmz080
3. Caminero, A., Nistal, E., Herran, A.R., Perez-Andres, J., Ferrero, M.A., Ayala, L.V., Vivas, S., Ruiz de Morales, J.M.G., Albillos, S.M., Casqueiro, F.J. (2015). Differences in gluten metabolism among healthy volunteers, coeliac disease patients and first-degree relatives. British Journal of Nutrition, 114, 1157-67.
4. Caminero, A., Herran, A.R., Nistal, E., Perez-Andres, J., Vaquero, L., Vivas, S., Ruiz de Morales, J.M.G., Albillos, S.M., Casqueiro, J. (2014). Diversity of the cultivable human gut microbiome involved in gluten metabolism: isolation of microorganisms with potential interest for coeliac disease. FEMS Microbiology Ecology, 88, 309-19.
5. Caminero, A., Nistal, E., Arias, L., Vivas, S., Comino, I., Real, A., Sousa, C., Ruiz de Morales, J.M., Ferrero, M.A., Rodriguez-Aparicio, L.B., Casqueiro, J. (2012). A gluten metabolism study in healthy individuals shows the presence of faecal glutenasic activity. European Journal of Nutrition, 51, 293-99.
6. Crockett, R., Le, P., Vodovotzet, Y. (2011). How Do Xanthan and Hydroxypropyl Methylcellulose Individually Affect the Physicochemical Properties in a Model Gluten-Free Dough? Journal of Food Science, 76(3), 274-282.
7. Czaja-Bulsa, G., Bulsa, M. (2017). What Do We Know Now about IgE-Mediated Wheat Allergy in Children? Nutrients, 9(35), doi:10.3390/nu9010035.
8. Delcour, J.A., Joye, I.J., Pareyt, B., Wilderjans, E., Brijs, K., Lagrain, B. (2012). Wheat Gluten Functionality as a Quality Determinant in Cereal-Based Food Products. Annual Review of Food Science and Technology, 3, 469–92.

9. Di Cagno, R., Angelis, M.D., Lavermicocca, P., Vincenzi, M.D., Giovannini, C., Faccia, M., Gobbetti, M. (2002). Proteolysis by Sourdough Lactic Acid Bacteria: Effects on Wheat Flour Protein Fractions and Gliadin Peptides Involved in Human Cereal Intolerance. Applied and Environmental Microbiology, 68(2), 623–633.
10. Diaz-Amigo, C., Yeung, J.M. (2010). Critical Evaluation of Uncertainties of Gluten Testing: Issues and Solutions for Food Allergen Detection. Pathogens and Toxins in Foods: Challenges and Interventions. Edited by V.K. Juneja and J.N. Sofos. 2010 ASM Press, Washington, DC
11. Dunaevsky, Y.E., Tereshchenkova, V.F., Belozersky, M.A., Filippova, I.Y., Oppert, B., Elpidina, E.N. (2021). Effective Degradation of Gluten and Its Fragments by Gluten-Specific Peptidases: A Review on Application for the Treatment of Patients with Gluten Sensitivity. Pharmaceutics, 13, 1603. https://doi.org/10.3390/pharmaceutics13101603.
12. DuPont, F.M., Chan, R., Lopez, R., Vensel, W.H. (2005). Sequential Extraction and Quantitive Recovery of Gliadins, Glutenins, and Other Proteins from small samples of wheat flour. Journal of Agriculture and Food Chemistry, 53, 1575-84.
13. Dziuba, M, Nalecz, D., Szerszunowicz, I., Waga, J. (2014). Proteomic Analysis of Wheat α/A- and β-Gliadins. Czech. Journal of Food Sciences, 32(5), 437-42.
14. El-Salhy, M., Hatlebakk, J.G., Gilja, O.H., Hausken, T. (2015). The relation between celiac disease, nonceliac gluten sensitivity and irritable bowel syndrome. Nutrition Journal, 14, 92.
15. Fasano, A., Sapone, A., Zevallos, V., Schuppan, D. (2015). Nonceliac Gluten Sensitivity, Gastroenterology,148, 1195–1204.
16. Gibert, A., Espadaler, M., Canela, M.A., Sanchez, A., Vaque, C., Rafecas, M. (2006). Consumption of gluten-free products: should the threshold value for trace amounts of gluten be at 20, 100 or 200 p.p.m.? European Journal of Gastroenterology & Hepatology, 18(11), 1187-1195.
17. Gómez M, Sciarini LS. (2015). Gluten-Free Bakery Products and Pasta. In Arranz E, Fernández-Bañares F, Rosell CM, Rodrigo L, Peña AS, editors. Advances in the Understanding of Gluten Related Pathology and the Evolution of Gluten-Free Foods. Barcelona, Spain: OmniaScience. p. 565-604.
18. Igci, N., Sharafi, P., Demiralp, D.O., Demiralp, C.O., Yuce, A., Dokmeci, S. (2017). Application of Fourier transform infrared spectroscopy to biomolecular profiling of cultured fibroblast cells from Gaucher disease patients: A preliminary investigation. Advances in Clinical and Experimental Medicine, 26(7), 1053–1061.
19. Jönsson, T., Memon, A.A., Sundquist, K., Sundquist, J., Olsson, S., Nalla, A., Bauer, M., Linse, S. (2015). Digested wheat gluten inhibits binding between leptin and its receptor. BMC Biochemistry, 16, 3.
20. Lagrain, B., Brunnbauer, M., Rombouts, I., Koehler, P. (2013). Identification of Intact High Molecular Weight Glutenin Subunits from the Wheat Proteome Using Combined Liquid Chromatography-Electrospray Ionization Mass Spectrometry. PLoS ONE 8(3), e58682. doi:10.1371/journal.pone.0058682.
21. Liu, W., Zhang, Y., Gao, X., Wang, K., Wang, S., Zhang, Y., He, Z., Ma, W., Yan, Y. (2012). Comparative proteome analysis of glutenin synthesis and accumulation in developing grains between superior and poor quality bread wheat cultivars. Journal of the Science of Food and Agriculture, 92, 106-115.
22. Matos, M., Rosell, C. (2014). Understanding gluten-free dough for reaching breads with physical quality and nutritional balance. Journal of the Science of Food and Agriculture, 95, 653–661.
23. Peraaho, M., Kaukinen,K., Paasikivi, K., Sievanen, H., Lohıniemi, S., Makis, M., Collin, P. (2003). Wheat-starch-based gluten-free products in the treatment of newly detected coeliac disease: prospective and randomized study. Alimentary Pharmacology and Therapeutics, 17, 587–594. Primo-Martin, C., Valera, R., Martianez-Anaya, M.A. (2003). Effect of Pentosanase and Oxidases on the Characteristics of Doughs and the Glutenin Macropolymer (GMP). Journal of Agricultural and Food Chemistry, 51, 4673-4679.
24. Rajpoot, P., Makharia, G.K. (2013). Problems and Challenges to Adaptation of Gluten Free Diet by Indian Patients with Celiac Disease. Nutrients, 5, 4869-4879
25. Ronda, F., Pérez-Quirce, S., & Villanueva, M. (2017). Chapter 12 - Rheological Properties of Gluten-Free Bread Doughs: Relationship With Bread Quality, Editor(s): J. Ahmed, P. Ptaszek, S. Basu, In Woodhead Publishing Series in Food Science, Technology and Nutrition, Advances in Food Rheology and Its Applications, Woodhead Publishing, Pages 297-334, ISBN 9780081004319, https://doi.org/10.1016/B978-0-08-100431-9.00012-7.
26. Secundo, F., Guerrier, N. (2005). ATR-FT/IR Study on the Interactions between Gliadins and Dextrin and Their Effects on Protein Secondary Structure. Journal of Agricultural and Food Chemistry, 53, 1757−1764.
27. Shewry, P.R., Halford, N.G., Belton, P.S., Tatham, A.S. (2002). The structure and properties of gluten: an elastic protein from wheat grain. Philosophical Transactions of the Royal Society of London. Series B, 357, 133-142.
28. Sivam, A.S., Sun-Waterhouse, D., Quek, S.Y., Perera, C.O. (2010). Properties of Bread Dough with Added Fiber Polysaccharides and Phenolic Antioxidants: A Review. Journal of Food Science, 75(8), 163-174.
29. Smerdel, B., Pollak, L., Novotni, D., Cukelj, N., Benkovic, M., Lusic, D., Curic, D. (2012). Improvement of gluten-free bread quality using transglutaminase, various extruded flours and protein isolates. Journal of Food and Nutrition, 51(4), 242–253.
30. Solé-Jamault, V., Davy, J., Cochereau, R., Boire, A., Larré, C., Denery-Papini, S. (2022). Optimization of large-scale purification of omega gliadins and other wheat gliadins, Journal of Cereal Science, 103, 103386, ISSN 0733-5210, https://doi.org/10.1016/j.jcs.2021.103386.
31. Stantiall, S.E., Serventi, L. (2017). Nutritional ans sensory challenges of glüten-free bakery products: a review. International Journal of Food Sciences and Nutrition, http://dx.doi.org/10.1080/09637486.2017.1378626
32. Volta, U., Caio,G., Tovoli, F., De Giorgio,R. (2013). Non-celiac gluten sensitivity: questions still to be answered despite increasing awareness. Cellular & Molecular Immunology, 10, 383–392.