ISSN 2308-4057 (Print),
ISSN 2310-9599 (Online)

Separation of gliadins from wheat flour by capillary gel electrophoresis: optimal conditions

Аннотация
Introduction. Gliadin proteins are one of the gluten fractions. They are soluble in alcoholic solution and divided into four groups (α + β, γ, ω1.2, and ω5-gliadins). In this paper gliadins were extracted from wheat flour, and optimal conditions for their separation were determined.
Study objects and methods. The separation was performed by capillary gel electrophoresis on Agilent apparatus, CE 7100 (a capillary with an inner diameter of 50 μm, a total length of 33 cm, and an effective length of 23.50 cm). In order to determine the optimal conditions, different solvent concentrations (50, 60, and 70% ethanol), capillary temperatures (20, 25, 30, 35, and 40°C), and electrode voltages (–14.5, –16.5, –17.5 and –18.5 kV) were applied. Migration time and relative concentration of each protein molecules within gliadin fractions in the electrophoregram were analysed using Agilent ChemStation Software.
Results and discussion. The optimal conditions for gliadin separation were: solvent 70% (v/v) ethanol, capillary temperature of 25°C, and electrode voltage of –16.5 kV. Under these conditions, the total proteins were indetified as Xav = 23.50, including α + β gliadin fraction (Xav = 7.50 and relative concentration RC = 28.29%), γ-gliadins (Xav = 5.00, RC = 26.66%), ω1.2-gliadins (Xav = 4.33, RC = 14.93%), and ω5-gliadins (Xav = 6.67, RC = 30.98%).
Conclusion. The results of the research can be of fundamental importance in the study of gluten proteins and the influence of technological procedures on their change and the possibility of reducing the allergic effect of gluten during processing.
Ключевые слова
Proteins , wheat , extraction , ethanol , electrophoresis , gluten
ФИНАНСИРОВАНИЕ
This work is a part of the research project “Analysis gluten proteins holders of allergic reactions by high performance liquid chromatography”, number 19/6-020/96123/18, financed by the Ministry of Scientific and Technological Development, Higher Education and Information Society Republic of Srpska.
СПИСОК ЛИТЕРАТУРЫ
  1. Shan L, Khosla C. Chemistry and biology of gluten proteins. Immunology, Endocrine and Metabolic Agents in Medicinal Chemistry. 2007;7(3):187–193. DOI: https://doi.org/10.2174/187152207780832397.
  2. Urade R, Sato N, Sugiyama M. Gliadins from wheat grain: an overview, from primary structure to nanostructures of aggregates. Biophysical Reviews. 2018;10(2):435–443. DOI: https://doi.org/10.1007/s12551-017-0367-2
  3. Wieser H. Chemistry of gluten proteins. Food Microbiology. 2007;24(2):115–119. DOI: https://doi.org/10.1016/j.fm.2006.07.004.
  4. Zhang Y, Luo G, Liu D, Wang D, Yang W, Sun J, et al. Genome-, transcriptome- and proteome wide analyses of the gliadin gene families in Triticum urartu. PloS ONE. 2015;10(7). DOI: https://doi.org/10.1371/journal.pone.0131559.
  5. Balakireva AV, Zamyatnin AA. Properties of gluten intolerance: Gluten structure, evolution, pathogenicity and detoxification capabilities. Nutrients. 2016;8(10). DOI: https://doi.org/10.3390/nu8100644.
  6. Wieser H. Relation between gliadin structure and coeliac toxicity. Acta Paediatrica. 1996;85(412):3–9. DOI: https://doi.org/10.1111/j.1651-2227.1996.tb14239.x.
  7. Qian Y, Preston K, Krokhin O, Mellish J, Ens W. Characterization of wheat gluten proteins by HPLC and MALDI TOF mass spectrometry. Journal of the American Society for Mass Spectrometry. 2008;19(10):1542–1550. DOI: https://doi.org/10.1016/j.jasms.2008.06.008.
  8. Lexhaller B, Colgrave ML, Scherf KA. Characterization and relative quantitation of wheat, rye, and barley gluten protein types by liquid chromatography-tandem mass spectrometry. Frontiers in Plant Science. 2019;10. DOI: https://doi.org/10.3389/fpls.2019.01530.
  9. Seilmeier W, Valdez I, Mendez E, Wieser H. Comparative investigations of gluten proteins from different wheat species II. Characterization of ω-gliadins. European Food Research and Technology. 2001;212(3):355–363. DOI: https://doi.org/10.1007/s002170000260.
  10. Horváth C. Storage proteins in wheat (Triticum aestivum L.) and the ecological impacts affecting their quality and quantity, with a focus on nitrogen supply. Journal of Agricultural and Environmental Sciences. 2014;1(2):57–76.
  11. Cebolla Á, Moreno ML, Coto L, Sousa C. Gluten immunogenic peptides as standard for the evaluation of potential harmful prolamin content in food and human specimen. Nutrients. 2018;10(12). DOI: https://doi.org/10.3390/nu10121927.
  12. Helmerhorst EJ, Zamakhchari M, Schuppan D, Oppenheim FG. Discovery of a novel and rich source of glutendegrading microbial enzymes in the oral cavity. PloS ONE. 2010;5(10). DOI: https://doi.org/10.1371/journal.pone.0013264.
  13. Shewry P. What is gluten – Why is it special? Frontiers in Nutrition. 2019;6. DOI: https://doi.org/10.3389/fnut.2019.00101.
  14. Grosch W, Wieser H. Redox reactions in wheat dough as affected by ascorbic acid. Journal of Cereal Science. 1999;29(1):1–16. DOI: https://doi.org/10.1006/jcrs.1998.0218.
  15. Srinivasan B, Focke-Tejkl M, Weber M, Pahr S, Baar A, Atreya R, et al. Usefulness of recombinant γ-gliadin 1 for identifying patients with celiac disease and monitoring adherence to a gluten-free diet. The Journal of Allergy and Clinical Immunology. 2015;136(6):1607–1618. DOI: https://doi.org/10.1016/j.jaci.2015.04.040.
  16. Daniel C, Triboi E. Effects of temperature and nitrogen nutrition on the grain composition of winter wheat: Effects on gliadin content and composition. Journal of Cereal Science. 2000;32(1):45–56. DOI: https://doi.org/10.1006/jcrs.2000.0313.
  17. Wieser H, Kieffer R. Correlations of the amount of gluten protein types to the technological properties of wheat flours determined on a micro-scale. Journal of Cereal Science. 2001;34(1):19–27. DOI: https://doi.org/10.1006/jcrs.2000.0385.
  18. Hurkman WJ, Tanaka CK, Vensel WH, Thilmony R, Altenbach SB. Comparative proteomic analysis of the effect of temperature and fertilizer on gliadin and glutenin accumulation in the developing endosperm and flour from Triticum aestivum L. cv. Butte 86. Proteome Science. 2013;11(1). DOI: https://doi.org/10.1186/1477-5956-11-8.
  19. Malvano F, Albanese D, Pilloton R, Di Matteo M. A new label-free impedimetric aptasensor for gluten detection. Food Control. 2017;79:200–206. DOI: https://doi.org/10.1016/j.foodcont.2017.03.033.
  20. Sirén H. Capillary electrophoresis in food analysis. In: Nollet LML, Toldra F, editors. Handbook of food analysis – Two Volume Set. Boca Raton: CRC Press; 2015. pp. 493–519. DOI: https://doi.org/10.1201/b18668.
  21. Lookhart G, Bean S. Separation and characterization of wheat protein fractions by high-performance capillary electrophoresis. Cereal Chemistry. 1995;72(6):527–532.
  22. Wenz C. Performance of commercially available gels for protein characterization by capillary gel electrophoresis with UV detection on the Agilent 7100 CE System. Application Note [Internet]. [cited 2020 Jul 1]. Available from: https://www.gimitec.com/file/5990-7976EN.pdf.
  23. Braud C, Devarieux R, Atlan A, Ducos C, Vert M. Capillary zone electrophoresis in normal or reverse polarity separation modes for the analysis of hydroxy acid oligomers in neutral phosphate buffer. Journal of hromatography B: Biomedical Sciences and Applications. 1998;706(1):73–82. DOI: https://doi.org/10.1016/s0378-4347(97)00468-4.
  24. Grujić R, Savanović D. Analysis of myofibrillar and sarcoplasmic proteins in pork meat by capillary gel electrophoresis. Foods and Raw Materials. 2018;6(2):421–428. DOI: https://doi.org/10.21603/2308-4057-2018-2-421-428.
  25. Bietz JA, Schmalzried E. Capillary electrophoresis of wheat gliadin: initial studies and application to varietal identification. LWT – Food Science and Technology. 1995;28(2):174–184. DOI: https://doi.org/10.1016/s0023-6438(95)91346-7.
Как цитировать?
Separation of gliadins from wheat flour by capillary gel electrophoresis: optimal conditions. Foods and Raw Materials, 2020, vol. 8, no. 2, pp. 411-421
DOI
http://doi.org/10.21603/2308-4057-2020-2-411-421
Издатель
Кемеровский государственный университет
htpps://kemsu.ru
ISSN
2308-4057 (Print) /
2310-9599 (Online)
О журнале