Improvement of Surface Functionalities, Including Allergenicity Attenuation, of Whole Buckwheat Protein Fraction by Maillard-Type Glycation with Dextran

  • Tazawa, Shigeru (Interdisciplinary Graduate School of Science and Technology, Shinshu University) ;
  • Katayama, Shigeru (Interdisciplinary Graduate School of Science and Technology, Shinshu University) ;
  • Hirabayashi, Masahiro (Department of Agriculture, Shinshu University) ;
  • Yamaguchi, Daiki (Department of Agriculture, Shinshu University) ;
  • Nakamura, Soichiro (Interdisciplinary Graduate School of Science and Technology, Shinshu University)
  • Received : 2014.05.22
  • Accepted : 2014.10.13
  • Published : 2014.12.31


The purpose of the current study was to determine the effects of the introduction of polysaccharide chains onto the molecular surface of buckwheat proteins on buckwheat protein surface functionality. The whole buckwheat protein fraction (WBP) was prepared using 50 mM phosphate buffer (pH 7.5) containing 0.5 M NaCl and covalently linked with 6 kDa, 17.5 kDa, 40 kDa, 70 kDa, or 200 kDa dextran by Maillard-type glycation through controlled dry-heating at $60^{\circ}C$ and 79% relative humidity for two weeks. Conjugation with 40 kDa dextran improved the water solubility and emulsifying properties of WBP without causing a serious loss of available lysine; 84.9% of the free amino groups were conserved. In addition, we found that the introduction of dextran chains onto the molecular surfaces of WBP attenuated the antigenicity of WBP.


  1. Javornik B, Kreft I. 1984. Characterization of buckwheat protein. Fagopyrum 4: 30-38.
  2. Tomotake H, Yamamoto N, Yanaka N, Ohinata H, Yamazaki R, Kayashita J, Kato N. 2006. High protein buckwheat flour suppresses hypercholesterolemia in rats and gallstone formation in mice by hypercholesterolemic diet and body fat in rats because of its low protein digestibility. Nutrition 22: 166-173.
  3. Tomotake H, Shimaoka I, Kayashita J, Yokoyama F, Nakajoh M, Kato N. 2000. A Buckwheat protein product suppresses gallstone formation and plasma cholesterol more strongly than soy protein isolate in hamsters. J Nutr 130: 1670-1674.
  4. Wieslander G. 1996. Review on buckwheat allergy. Allergy 51: 661-665.
  5. Yanagihara Y, Koda A. 1980. Immunopharmacological study of buckwheat hypersensitivity. Nihon Yakurigaku Zasshi 10: 184-188.
  6. Yano M, Nakamura R, Hayakawa S, Torii S. 1989. Purification and properties of allergenic proteins in buckwheat seed. Agric Biol Chem 53: 2387-2392.
  7. Matsumoto R, Fujino K, Nagata Y, Hashiguchi S, Ito Y, Aihara Y, Takahashi Y, Maeda K, Sugimura K. 2004. Molecular characterization of a 10-kDa buckwheat molecule reactive to allergic patients' IgE. Allergy 59: 533-538.
  8. Urisu A, Kondo Y, Morita Y, Wada E. 1994. Identification of a major allergen of buckwheat seeds by immunoblotting methods. Allergy Clin Immunol News 6: 151-155.
  9. Nair A, Adachi T. 1999. Immunodetection and characterization of allergenic proteins in common buckwheat (Fagopyrum esculentum). Plant Biotechnol 16: 219-224.
  10. Yoshimasu MA, Zhang JW, Hayakawa S, Mine Y. 2000. Electrophoretic and immunochemical characterization of allergenic proteins in buckwheat. Int Arch Allergy Immunol 123: 130-136.
  11. Yoshioka H, Ohmoto T, Urisu A, Mine Y, Adachi T. 2004. Expression and epitope analysis of the major allergenic protein Fag e 1 from buckwheat. J Plant Physiol 161: 761-767.
  12. Tanaka K, Matsumoto K, Akasawa A, Nakajima T, Nagasu T, Iikura Y, Saito H. 2002. Pepsin-resistant 16-kD buckwheat protein is associated with immediate hypersensitivity reaction in patients with buckwheat allergy. Int Arch Allergy Immunol 129: 49-56.
  13. Arita K, Babiker EE, Hiroyuki A, Kato A. 2001. Effect of chemical and genetic attachment of polysaccharides to proteins on the production of IgG and IgE. J Agric Food Chem 49: 2030-2036.
  14. Nakamura S, Suzuki Y, Ishikawa E, Yakushi T, Jing H, Miyamoto T, Hashizume K. 2008. Reduction of in vitro allergenicity of buckwheat Fag e 1 through the Maillard-type glycosylation with polysaccharides. Food Chem 109: 538-545.
  15. Marshall JJ, Rabinowitz ML. 1976. Preparation and characterization of a dextran-trypsin conjugate. J Biol Chem 251: 1081-1087.
  16. Nakamura S, Kato A, Kobayashi K. 1992. Bifunctional lysozyme-galactomannan conjugate having excellent emulsifying properties and bactericidal effect. J Agric Food Chem 40: 735-739.
  17. Kato A, Murata K, Kobayashi K. 1988. Preparation and characterization of ovalbumin-dextran conjugate having excellent emulsifying properties. J Agric Food Chem 36: 421-425.
  18. Nakamura S, Dokai K, Matsuura M, Saeki H. 2002. Food safety of functional neoglycoproteins prepared by covalent attachment of galactomannan to food proteins. Nutraceuticals & Food 7: 139-145.
  19. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
  20. Zacharius RM, Zell TE, Morrison JH, Woodlock JJ. 1969. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem 30: 148-152.
  21. Haynes R, Osuga DT, Feeney RE. 1967. Modification of amino groups in inhibitors of proteolytic enzymes. Biochemistry 6: 541-547.
  22. Pearce KM, Kinsella JE. 1978. Emulsifying properties of proteins: evaluation of a turbidimetric technique. J Agric Food Chem 26: 716-723.
  23. Okahata Y, Niikura K, Sugiura Y, Sawada M, Morii T. 1998. Kinetic studies of sequence-specific binding of GCN4-bZIP peptides to DNA strands immobilized on a 27-MHz quartzcrystal microbalance. Biochemistry 37: 5666-5672.
  24. Choi SM, Ma CY. 2006. Extraction, purification and characterization of globulin from common buckwheat (Fagopyrum esculentum Moench) seeds. Food Res Int 39: 974-981.
  25. Sarwat F, Ul Qader SA, AmanA, Ahmed N. 2008. Production and characterization of a unique dextran from an indigenous Leuconostoc mesenteroides CMG713. Int J Biol Sci 4: 379-386.

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