Production of Neoglycoprotein with Neutralizing Activity to Bacterial Toxins from Cheese Whey and Its Characteristic Analysis.
从奶酪乳清中制备具有细菌毒素中和活性的新糖蛋白及其特性分析。
基本信息
- 批准号:08660326
- 负责人:
- 金额:$ 1.6万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:1996
- 资助国家:日本
- 起止时间:1996 至 1997
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
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.
a.以甜干酪乳清为原料,采用12%三氯乙酸(TCA)直接萃取法制备酪蛋白糖肽(CGP)。唾液乳糖通过三种不同的化学或酶促技术,尝试了N-乙酰神经氨酸(NeuAc)α 2-x-半乳糖基β 1 - 4葡萄糖)对CGP的作用,最后是非糖蛋白(neoCGP)c.引入的糖链经碱性硼氢化物处理裂解后通过甲基化和NMR分析来分析其化学结构。d.利用生物素化聚合物探针建立了一种新的检测霍乱毒素中和活性的方法。首先,用霍乱毒素B亚基包被聚苯乙烯珠。第二,珠与样品反应,然后与生物素化NeuAc探针孵育。第三,在与HRP-链亲和素孵育后,将珠显色。如果样品对霍乱毒素具有中和活性,则与空白相比显色水平降低。e.通过使用开发的新评价方法,与去唾液酸CGP相比,实际上证实了neoCGP的中和活性增加。
项目成果
期刊论文数量(0)
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$ 1.6万 - 项目类别:
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