Affinity Preparation of Sialylsulfo-glycopeptides from Hen Ovomucin and Their Characteristic Analysis

母鸡卵粘蛋白唾液酸磺基糖肽的亲和制备及其特性分析

• 批准号: 06660333
• 负责人: SAITO Tadao
• 金额: $ 1.34万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06660333/
• 关键词: Chicken egg; Egg white proteins; Ovomucin; Sialic acid; Glycopeptides; Affinity chromatography; Neutralizing activity; Cholera toxin; 総合糖鎖; 化学構造

a. Sugar chains were released from hen ovomucin by alkaline borohydride reaction. Neutral and acidic sugar chains were isolated by ion-exchange chromatography. Neutral sugar chains were purified Bio-Gel P-4 gel filtration. Acidic sugar chains were purified by ion-exchange and high performance liquid chromatographies. The chemical structures of sugar chains were analyzed by methylation and nuclear magnetic resonance analysis.b. Three novel neutral sugar chains (mono-, di- and trisaccharide) and three novel acidic sugar chains (tetra-, penta- and hexasaccharide) were finally clarified.c. Ovomucin was digested by actinase E.Neutral and acidic glycopeptides (sialyl- and sialylsulfo-glycopeptides) were separated by lysozyme-affinity chromatography which was developed in our previous paper.d. A new assay for evaluation of neutralizing activity against cholera toxin by using polystirene beads. This system was more high sensitive compared to microplate method.e. Sialyl- and sialylsulfo-glycopeptide isolated from hen ovomucin by lysozyme-affinity chromatography showed high neutralizing activity against cholera toxin.

a.通过碱性硼氢化物反应从鸡蛋粘蛋白中释放糖链。通过离子交换色谱分离中性和酸性糖链。中性糖链经Bio-Gel P-4凝胶过滤纯化。通过离子交换和高效液相色谱法纯化酸性糖链。通过甲基化和核磁共振分析来分析糖链的化学结构。最终确定了三种新的中性糖链（单糖、二糖和三糖）和三种新的酸性糖链（四糖、五糖和六糖）。卵粘蛋白经放线菌素E酶解后，用我们以前建立的溶菌酶亲和层析法分离了中性和酸性糖肽（唾液酸-和唾液酸磺基-糖肽）。一种新的评价聚苯乙烯珠中和霍乱毒素活性的方法。与微孔板法相比，该系统具有更高的灵敏度。用溶菌酶亲和层析法从鸡卵粘蛋白中分离得到唾液酸糖肽和唾液酸磺基糖肽，它们对霍乱毒素具有很高的中和活性。

项目成果

Tadao Saito et al.: "The Chemical Structure of Novel Neutral Sugar Chains Isolated from Hen Ovomucin." Biosci. Biotech. Biochem.(1996)

Tadao Saito 等人：“从母鸡卵粘蛋白中分离出的新型中性糖链的化学结构”。

Tadao Saito et al.: "Mass Production of Sialylsulfo-glycopeptides from Hen Ovomucin and Their Neutralizing Activity for Cholera Toxin." Biosci. Biotech. Biochem.(1996)

Tadao Saito 等人：“从母鸡卵粘蛋白中大量生产唾液酸磺基糖肽及其对霍乱毒素的中和活性”。

Tadashi Nakamura et al.: "Isolation of a New Minor Protein (Ovofactor-1), Which Has a Cell Growth Promoting Activity, from Hen's Egg White by Heparin Affinity Chromatography." Biosci. Biotech. Biochem.59. 1946-1948 (1995)

Tadashi Nakamura 等人：“通过肝素亲和色谱法从鸡蛋清中分离出一种具有细胞生长促进活性的新型次要蛋白（Ovofactor-1）。”

Tadao,Saito et al.: "Fraction of Glycopeptides form Bovine K-Casein and Hen Ovomucin Via Lysozyme Affinity Chromatography" Anim.Sci.Technol.(Jpn). 65. 624-630 (1994)

Tadao, Saito 等人：“通过溶菌酶亲和色谱法从牛 K-酪蛋白和母鸡卵粘蛋白中提取糖肽”Anim.Sci.Technol.（日本）。

Tadao Saito et al.: "The Chemical Structure of Novel Sialysulfo-Sugar Chains Isolated from Hen Ovomucin." Biosci. Biotech. Biochem.(1996)

Tadao Saito 等人：“从母鸡卵粘蛋白中分离出的新型唾液酸磺基糖链的化学结构”。