Change of the Content of an Allergenic Protein in Soybean during Natto Fermentation

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  • 納豆発酵過程における大豆アレルゲン蛋白質の変動
  • ナットウ ハッコウ カテイ ニ オケル ダイズ アレルゲン タンパクシツ ノ

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In this study, in order to develop a hypollergenic soybean product, the change of the content of an allergenic protein, Gly m Bd 30K, in soybean during natto fermentation in a natto manufacturing factory was investigated by the immunological methods with anti-Gly m Bd 30K antiserum. For the preparation of the anti-Gly m Bd 30K antiserum, was purified from the defatted soybean meal by extraction, isoelectric precipitation at pH6.4, ammonium sulfate fractionation (40% saturation), Sephacryl S-400 and Cellulofine GCL-1000sf gel filtrations (Fig. 1), and HiTrap Q ion-exchange chromatography (Fig. 2A). The purified Gly m Bd 30K was separated into two bands with the molecular weights of approximately 33,000 and 31,000, respectively on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Fig. 2B). Both bands were identified as Gly m Bd 30K by the N-terminal amino acid sequence analysis. Then, the carboxymethylated Gly m Bd 30K was immunized to two rabbits and after eight weeks, anti-Gly m Bd 30K antisera were obtained. By immunoblotting with the anti-Gly m Bd 30K antiserum, the allergenic protein was detected in Hikiwari soybean fermented for 0-9hrs, but not in Hikiwari soybean fermented for 10-17hrs (Fig. 3). The contents of Gly m Bd 30K in both Hikiwari and Kotsubu natto were reduced to 1-5% of that of the unfermented soybean after the fermentation for 10hrs or longer(Fig. 4). By an immunoblotting technique with the serum of a soybean-sensitive patient with atopic dermatitis, one of the major allergenic proteins of soybean with the molecular weight of approximately 69,000, could not be detected in Hikiwari natto fermented for both 10 and 17hrs (Fig. 5). From these results, it was suggested that Hikiwari natto fermented for 10-17hrs in the natto manufacturing factory was suitable for the preparation of the hypoallergenic soybean product.

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