Production of Neoglycoprotein with Neutralizing Activity to Bacterial Toxins from Cheese Whey and Its Characteristic Analysis.

点击翻译按钮获取中文摘要

a.The new preparation method for caseinoglycopeptides (CGP) from sweet cheese whey by direct extraction with 12% TCA was developed.b.The introduction of an acidic trisaccharide ; sialyllactose (N-acetylneuraminil (NeuAc) alpha2-x-galactosylbeta1-4glucose) was tried against CGP by three different chemical or enzymatic technique and finally noeglycoprotein (neoCGP) was produced.c.The chemical structure of the introduced sugar chains was analyzed by methylation and NMR analysis after cleavage by alkaline borohydride treatment.d.A new detection method for neutralizing activity to Cholera toxin was developed by using biotinated-polymer probe. First, polystirene beads were coated with Cholera toxin B-subunit. Second, the beads was reacted with the sample and then incubated with biotinated NeuAc-probe. Third, the beads was color-developed after incubation with HRP-streptoavidin. If the sample has neutralizing activity to Cholera toxin, the coloration level is decreased compared to blank.e.The increase of the neutralizing activity of neoCGP was actually confirmed compared to asialoCGP by using the developed new evaluation method.