裙帶菜褐藻糖膠的提取、純化及組分研究

食品科學論文編號:SP129  論文字數:9839,頁數:30

目錄
中文摘要 I
英文摘要 II
目錄 Ⅲ
1 緒論 1
1.1 裙帶菜簡介 1
1.2 褐藻糖膠簡介 2
1.3  褐藻糖膠的研究方法 2
2 實驗部分 5
2.1  材料 5
2.1.1  裙帶菜(Undaria pinnatifida) 5
2.1.2  試劑和設備 5
2.2  水提法 7
2.3  酸提法 8
2.4  葡萄糖含量標準曲線的繪制 8
2.4.1  試劑 8
2.4.2  標準曲線的繪制 9
2.4.3  粗多糖含量的測定 9
2.5  粗多糖的純化 9
2.6  Sevag法除褐藻糖膠中蛋白質 9
2.7  考馬思亮藍法測定蛋白質含量標準曲線的繪制 10
2.7.1  試劑 10
2.7.2  蛋白質含量標準曲線的繪制 10
2.8  褐藻糖膠的層析法分離 10
2.8.1  試劑 10
2.8.2  層析方法 11
2.9  硫酸根含量標準曲線的繪制 11
2.9.1  試劑 11
2.9.2  硫酸根含量標準曲線的繪制 12
2.9.3  換算因子（f）的測定 12
2.9.4  褐藻糖膠中硫酸根含量的測定 12
2.10  氣相色譜法測定褐藻糖膠中單糖組成 13
2.10.1  標準單糖衍生化 13
2.10.2  褐藻糖膠水解和衍生化 13
2.10.3  氣相色譜條件 13
3  結果與討論 14
3.1  水提法和酸提法的比較 14
3.2  CaC12法純化粗多糖 14
3.3  褐藻糖膠除蛋白質的檢測 14
3.4  柱層析分離 15
3.5  褐藻糖膠中單糖組成分析 17
3.5.1  標準單糖氣相色譜圖 17
3.5.2  褐藻糖膠組分氣相色譜圖 18
4 總結與展望 21
4.1  試驗總結 21
4.2  展望 21
致謝 23
參考文獻 24

摘要：褐藻糖膠是存在于褐藻中的硫酸酯多糖，具有抗凝血、抗病毒等多種生理活性。本實驗采用甲醇對新鮮裙帶菜進行預處理，分別用水提法和酸提法提取多糖，采用不同濃度乙醇進行分級沉淀。用蒽酮-硫酸法測定各級沉淀中多糖的含量，結果多糖基本上存在60%乙醇濃度時的沉淀中。提取的粗多糖樣品采用CaCl2法進行純化，得到褐藻糖膠。褐藻糖膠采用Sevag法去除蛋白質后，上DEAE-Sepharose Fast Flow離子交換層析進行分級，用蒽酮硫酸法檢測洗脫峰，得到五個峰，對應為F1, F2, F3, F4, F5五個組分。用硫酸鋇比濁法測定五個組分的硫酸根含量分別為F1為18.3%；F2,為19.2%；F3為30.1%；F4為38.4%；F5為32.96%。多糖組分三氟乙酸水解后，對水解產物進行糖腈乙酸酯衍生化后，氣相色譜分析確定F1主要含有甘露糖和半乳糖；F2主要含有鼠李糖，巖藻糖，木糖，甘露糖，葡萄糖和半乳糖；F3主要含有巖藻糖；F4主要含有巖藻糖和半乳糖；F5主要含有巖藻糖和半乳糖。

關鍵詞：裙帶菜；巖藻聚糖；硫酸根；單糖組成

Abstract: Fucoidan is a sulfated polysaccharide from brown seaweed, which has a variety of biological activities, such as anticoagulant activities and antiviral activities. In this paper, Fucoidan was extracted from Undaria pinnatifida pretreated with methanol by acid method and aqueous method, and separated by fractional precipitation with alcohol. The fucoidan was found in the deposition when the consistence of ehthol was 60%. The crude fucoidan was purified by CaCl2 method. Then, the protein in fucoidan was removed by sevag way. DEAE-Sepharose Fast Flow column chromatography was used to isolate purification fucoidan, and F1、F2、F3、F4、F5 were obtained. The content of sulfate and the composition of the properties were analyzed. The content of sulfate was as follow: F1, 18.3%; F2, 19.2%; F3, 30.1%; F4, 38.4%; F5, 32.96%. The monosaccharide composition of the properties was identified with trifluoroacetic acid by the method of aldononitrile acetate precolumn-derivatizaion of Gas chromatography. The composition was as follow: F1, mannose and galactose; F2, rhamnose、fucose、xylose、mannose、glaucose and galactose; F3, fucose; F4, fucose and galactose; F5, fucose and galactose.

Keywords: Undaria pinnatifida; Fucoidan; sulfate; monosaccharide composition