Nutr Res Pract.  2014 Jun;8(3):278-283.

Effects of enzymatic hydrolysis of buckwheat protein on antigenicity and allergenicity

Affiliations
  • 1Department of Food Science and Technology, 52, Ewhayeodae-gil, Seodaemun-gu, Seoul 120-750, Korea. ssoh71@ewha.ac.kr
  • 2Environmental Health Center for Atopic Diseases, Samsung Medical Center, Seoul 135-710, Korea.
  • 3Korea Food Research Institute, Gyeonggi 463-746, Korea. ssoh71@ewha.ac.kr

Abstract

BACKGROUND/OBJECTIVES
Due to its beneficial health effects, use of buckwheat has shown a continuous increase, and concerns regarding the allergic property of buckwheat have also increased. This study was conducted for evaluation of the hydrolytic effects of seven commercial proteases on buckwheat allergens and its allergenicity.
MATERIALS/METHODS
Extracted buckwheat protein was hydrolyzed by seven proteolytic enzymes at individual optimum temperature and pH for four hours. Analysis was then performed using SDS-PAGE, immunoblotting, and competitive inhibition ELISA (ciELISA) with rabbit antiserum to buckwheat protein, and direct ELISA with pooled serum of 21 buckwheat-sensitive patients.
RESULTS
Alkaline protease, classified as serine peptidase, was most effective in reducing allergenicity of buckwheat protein. It caused decomposition of the whole buckwheat protein, as shown on SDS-PAGE, and results of immunoblotting showed that the rabbit antiserum to buckwheat protein no longer recognized it as an antigen. Allergenicity showed a decrease of more than 50% when pooled serum of patients was used in ELISA. Two proteolytic enzymes from Aspergillus sp. could not hydrolyze buckwheat allergens effectively, and the allergenicity even appeared to increase.
CONCLUSIONS
Serine-type peptidases appeared to show a relatively effective reduction of buckwheat allergenicity. However, the antigenicity measured using rabbit antiserum did not correspond to the allergenicity measured using sera from human patients. Production of less allergenic buckwheat protein may be possible using enzymatic hydrolysis.

Keyword

Buckwheat; allergy; enzymatic hydrolysis

MeSH Terms

Allergens
Aspergillus
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Fagopyrum*
Humans
Hydrogen-Ion Concentration
Hydrolysis*
Hypersensitivity
Immunoblotting
Peptide Hydrolases
Serine
Allergens
Peptide Hydrolases
Serine

Figure

  • Fig. 1 SDS-PAGE analysis of enzymatic hydrolyzed buckwheat protein (A) and immunoblotting of the hydrolyzed samples with rabbit antiserum (B). M: Molecular weight marker, BW: buckwheat protein, AK: Alkaline protease, FLA: Flavourzyme, BRO: Bromelain, COL: Collupulin, PRO: Protamex, GC: GC106, PAP: Papain

  • Fig. 2 Allergenicity of enzymatic hydrolyzed buckwheat protein measured with pooled serum of buckwheat-sensitive patients. Patient indicates pooled serum of buckwheat-sensitive patients, and negative indicates sera from healthy and non-atopic individuals. BW: buckwheat native protein, AK: Alkaline protease, FLA: Flavourzyme, BRO: Bromelain, COL: Collupulin, PRO: Protamex, GC: GC106, PAP: Papain. Error bars indicate standard deviation.


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