Comparable efficiency of different extraction protocols for wheat and rye prolamins

Authors

  • Peter Socha Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Research Centre AgroBioTech, Tr. A. Hlinku 2, 949 76 Nitra
  • Marián Tomka Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Biochemistry and Biotechnology, Tr. A. Hlinku 2, 949 76 Nitra
  • Kvetoslava Kačmárová Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Biochemistry and Biotechnology, Tr. A. Hlinku 2, 949 76 Nitra
  • Blažena Lavová Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Biochemistry and Biotechnology, Tr. A. Hlinku 2, 949 76 Nitra
  • Eva Ivanišová Slovak University of Agriculture in Nitra, Faculty of Biotechnology and Food Sciences, Department of Plant Products Storage and Processing, Tr. A. Hlinku 2, 949 76 Nitra
  • Barbara Mickowska University of Agriculture, Faculty of Food Technology, Malopolska Centre of Food Monitoring, Balicka 122, 30-149 Krakow
  • Dana Urminská Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra, Institute of Chemistry, Slovak Academy of Sciences, Center for White-Green Biotechnology, Tr. A. Hlinku 2, 949 76 Nitra

DOI:

https://doi.org/10.5219/540

Keywords:

extraction, prolamins, wheat, rye

Abstract

The identification and quantification of cereal storage proteins is of interest of many researchers. Their structural or functional properties are usually affected by the way how they are extracted. The efficiency of extraction process depends on the cereal source and working conditions. Here, we described various commonly used extraction protocols differing in the extraction conditions (pre-extraction of albumins/globulins, sequential extraction of individual protein fractions or co-extraction of gluten proteins, heating or non-heating, reducing or non-reducing conditions). The total protein content of all fractions extracted from commercially available wheat and rye flours was measured by the Bradford method. Tris-Tricine SDS-PAGE was used to determine the molecular weights of wheat gliadins, rye secalins and high-molecular weight glutelins which are the main triggering factors causing celiac disease. Moreover, we were able to distinguish individual subunits (α/β-, γ-, ω-gliadins and 40k-γ-, 75k-γ-, ω-secalins) of wheat/rye prolamins. Generally, modified extraction protocols against classical Osborne procedure were more effective and yields higher protein content in all protein fractions. Bradford measurement led into underestimation of results in three extraction procedures, while all protein fractions were clearly identified on SDS-PAGE gels. Co-extraction of gluten proteins resulted in appearance of both, low-molecular weight fractions (wheat gliadins and rye secalins) as well as high-molecular weight glutelins which means that is not necessary to extract gluten proteins separately. The two of three extraction protocols showed high technical reproducibility with coefficient of variation less than 20%. Carefully optimized extraction protocol can be advantageous for further analyses of cereal prolamins. 

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References

Blum, H., Beier, H., Gross, H. J. 1987. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels, Electrophoresis, vol. 8, p. 93-99. https://doi.org/10.1002/elps.1150080203 DOI: https://doi.org/10.1002/elps.1150080203

van den Broeck, H. C., America, A. H. P., Smulders, M. J. M., Bosch, D., Hamer, R. J., Gilissen, L. J. W. J., van der Meer, I. M. 2009. A modified extraction protocol enables detection and quantification of celiac-disease related gluten proteins from wheat. Journal of Chromatography B, vol. 877, p. 975-982. https://doi.org/10.1016/j.jchromb.2009.02.035 DOI: https://doi.org/10.1016/j.jchromb.2009.02.035

van den Broeck, H. C., de Jong, H. C., Salentijn, E. M. J., Dekking, L., Bosch, D., Hamer, R. J., Gilissen, L. J. W. J., van der Meer, I. M., Smulders, M. J. M. 2010. Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease, Theoretical and Applied Genetics, vol. 121, no. 8, p. 1527-1539. https://doi.org/10.1007/s00122-010-1408-4 DOI: https://doi.org/10.1007/s00122-010-1408-4

van den Broeck, H. C., Gilissen, L. J. W. J., Smulders, M. J. M., van der Meer, I. M., Hamer, R. J. 2011. Dough quality of bread wheat lacking α-gliadins with celiac disease epitopes and addition of celiac-safe avenins to improve dough quality. Journal of Cereal Sciences, vol. 53, p. 206-216. https://doi.org/10.1016/j.jcs.2010.12.004 DOI: https://doi.org/10.1016/j.jcs.2010.12.004

Ciclitira, P. J., Ellis, H. J., Lundin, K. E. A. 2005. Gluten-free diet - what is toxic? Best Practice & Research Clinical Gastroenterology, vol. 19, no. 3, p. 359-371. https://doi.org/10.1016/j.bpg.2005.01.003 DOI: https://doi.org/10.1016/j.bpg.2005.01.003

DuPont, F. M., Chan, R., Lopez, R., Vensel, W. H. 2005. Sequential extraction and quantitative recovery of gliadins, glutenins, and other proteins from small samples of wheat flour. Journal of Agricultural and Food Chemistry, vol. 53, p. 1575-1584. https://doi.org/10.1021/jf048697l DOI: https://doi.org/10.1021/jf048697l

van Eckert, R., Bond, J., Rawson, P., Klein, Ch. L., Stern, M., Jordan, T. W. 2010. Reactivity of gluten detecting monoclonal antibodies to a gliadin reference material. Journal of Cereal Sciences, vol. 51, no. 2, p. 198-204. https://doi.org/10.1016/j.jcs.2009.11.012 DOI: https://doi.org/10.1016/j.jcs.2009.11.012

Gellrich, C., Schieberle, P., Wieser, H. 2003. Biochemical characterization and quantification of the storage protein (secalin) types in rye flour. Cereal Chemistry, vol. 80, no. 1, p. 102-109. https://doi.org/10.1094/CCHEM.2003.80.1.102 DOI: https://doi.org/10.1094/CCHEM.2003.80.1.102

Gregorini, A., Colomba, M., Ellis, H. J., Ciclitira, P. J. 2009. Immunogenicity characterization of two ancient wheat α-gliadin peptides related to coeliac disease. Nutrients, vol. 1, no. 2, p. 276-290. https://doi.org/10.3390/nu1020276 DOI: https://doi.org/10.3390/nu1020276

Hybenova, E., Štofirová, J., Mikulajová, A. 2013. Celiac disease and gluten-free diet. Potravinarstvo, vol. 7, no. 1, p. 95-100. https://doi.org/10.5219/276 DOI: https://doi.org/10.5219/276

Kaukinen, K., Lindfors, K., Mäki, M. 2014. Advances in the treatment of coeliac disease: an immunopathogenic perspective. Nature Reviews Gastroenterology & Hepatology, vol. 11, p. 36-44. PMid:23917697 DOI: https://doi.org/10.1038/nrgastro.2013.141

Kruger, J. E., Marchylo, B. A., Hatcher, D. 1988. Preliminary assessment of a sequential extraction scheme for evaluating quality by reversed-phase high-performance liquid chromatography and electrophoretic analysis of gliadins and glutenins. Cereal Chemistry, vol. 65, no. 3, p. 208-214.

Mamone, G., Picariello, G., Addeo, F., Ferranti, P. 2011. Proteomic analysis in allergy and intolerance to wheat products. Expert Review of Proteomics, vol. 8, no. 1, p. 95-115. https://doi.org/10.1586/epr.10.98 DOI: https://doi.org/10.1586/epr.10.98

Mickowska, B., Socha, P., Urminská, D., Cieślik, E. 2012. The comparison of prolamins extracted from different varieties of wheat, barley, rye and triticale species: amino acid composition, electrophoresis and immunodetection, Journal of Microbiology, Biotechnology and Food Sciences, vol. 1, no. 4, p. 742-752. ISSN 1338-5178.

Osborne, T. B. 1924. The Vegetable Proteins, 2nd ed., Longmans Green and Co., London, UK. 154 p.

Schägger, H., von Jagow, G. 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analytical Biochemistry, vol. 166, no. 2, p. 368-379. https://doi.org/10.1016/0003-2697(87)90587-2 DOI: https://doi.org/10.1016/0003-2697(87)90587-2

Shewry, P. R. 2004. Improving the protein content and quality of temperate cereals: wheat, barley and rye. In Çakmak, I. and Welch, R. M. Impacts of Agriculture on Human Health and Nutrition. EOLSS Publishers Co Ltd. : Oxford, UK. ISBN 978-1848265448.

Singh, N. K., Shepherd, K. W., Cornish, G. B. 1991. A simplified SDS-PAGE procedure for separating LMW subunits of glutenin. Journal of Cereal Science, vol. 14, no. 3, p. 203-208. https://doi.org/10.1016/S0733-5210(09)80039-8 DOI: https://doi.org/10.1016/S0733-5210(09)80039-8

Weiss, W., Vogelmeier, C., Görg, A. 1993. Electrophoretic characterization of wheat grain allergens from different cultivars involved in bakers' asthma. Electrophoresis, vol. 14, no. 8, p. 805-816. https://doi.org/10.1002/elps.11501401126 DOI: https://doi.org/10.1002/elps.11501401126

Wieser, H. 1998. Investigations on the extractability of gluten proteins from wheat bread in comparison with flour. Zeitschrift für Lebensmitteluntersuchung und - Forschung A, vol. 207, no. 2, p. 128-132. https://doi.org/10.1007/s002170050306 DOI: https://doi.org/10.1007/s002170050306

Wieser, H. 2007. Chemistry of gluten proteins. Food Microbiology, vol. 24, no. 2, p. 115-119. https://doi.org/10.1016/j.fm.2006.07.004 DOI: https://doi.org/10.1016/j.fm.2006.07.004

Wieser, H., Koehler, P. 2008. The biochemical basis of celiac disease. Cereal Chemistry, vol. 85, no. 1, p. 1-13. https://doi.org/10.1094/CCHEM-85-1-0001 DOI: https://doi.org/10.1094/CCHEM-85-1-0001

Zingone, F., Capone, P., Ciacci, C. 2010. Celiac disease: Alternatives to a gluten free diet. World Journal of Gastrointestinal Pharmacology and Therapeutics, vol. 1, no. 1, p. 36-39. PMid:21577293 DOI: https://doi.org/10.4292/wjgpt.v1.i1.36

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Published

2016-01-24

How to Cite

Socha, P. ., Tomka, M. ., Kačmárová, K. ., Lavová, B. ., Ivanišová, E. ., Mickowska, B. ., & Urminská, D. . (2016). Comparable efficiency of different extraction protocols for wheat and rye prolamins. Potravinarstvo Slovak Journal of Food Sciences, 10(1), 139–144. https://doi.org/10.5219/540

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