Purification and Characterization of the Lectins from Mushroom Flammulina velutipes

팽이버섯으로부터 Lectin의 정제와 특성

  • Kim, Hyung-Suk (Graduate School of Biotechnology, Korea University) ;
  • Son, Seung-Yeol (Dept. of Microbiology, College of Natural Sciences, Dankook University) ;
  • Hwang, Se-Young (Graduate School of Biotechnology, Korea University) ;
  • Hong, Bum-Shik (Graduate School of Biotechnology, Korea University)
  • 김형석 (고려대학교 생명공학원) ;
  • 손승렬 (단국대학교 미생물학과) ;
  • 황세영 (고려대학교 생명공학원) ;
  • 홍범식 (고려대학교 생명공학원)
  • Published : 1999.11.30

Abstract

Two Lectins, designated FVL-1 and FVL-2, were isolated and purified from the fruiting bodies of edible mushroom Flammulina veluripes using ammonium sulfate fractionation, ethanol treatment, DEAE-TOYPEARL ion-exchange column chromatography, and TSK-Gel HW-55F column chromatography. Specific activity increased 18 folds for FVL-1 and 7.9 folds for FVL-2 from ethanol treated sample. SDS-PAGE of FVL-1 and FVL-2 gave apparent molecular mass of 10.6 kDa and 37 kDa, respectively. FVL-2 agglutinated all type of human erythrocytes (A, B, AB, and O). However, FVL-1 agglutinated more human erythrocyte type O than type A, B, and AB. The hemagglutination activities of the FVL-1 were effectively inhibited by bovine submaxillary and porcine stomach mucins(BSM and PSM), fetuin, asialofetuin and cations, such as $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Mn^{2+}$ and $Fe^{2+}$. However, FVL-2 was not inhibited by any cations. The hemagglutination activities of the two lectins were not inhibited by the sugar, such as lactose, galactose and sugar derivatives. FVL-1 and FVL-2 were stable at pH $5{\sim}11$ and pH $4{\sim}7$, respectively. FVL-1 was stable below $55^{\circ}C$ and FVL-2 was below $45^{\circ}C$.