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

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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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