【作者】 周峙苗；

【导师】 孙培龙；

【作者基本信息】 浙江工业大学 ， 生物化工， 2004， 硕士

【摘要】 羊栖菜多糖(简称SFP)是从马尾藻科植物羊栖菜(Sargassum fusiforme(Harv.)Setchel)中提取得到的，具有抗肿瘤、抗凝血、降血糖、提高肌体免疫功能等生理活性。本文研究用水提法、酸提法、酶法提取羊栖菜多糖，考虑到提取温度、时间、加水量、加酶量等因素的影响，用正交试验优化酶法提取条件。酶法提取的最适温度为50℃，最适pH值为3.0，提取时间为3hr，加酶量为纤维素酶(16000 U／g)8％，果胶酶(20000 U／g)4％。为适应工业化生产需要，确定水提法为煮沸提取时间3hr，提取液pH值为3.0，固液比为1：40。用酶法结合高温浸提多糖提取的工艺条件是：加纤维素酶量为(16000U／g)8％，加果胶酶量为(20000 U／g)4％，pH3.0，酶解时间为3hr，酶解温度为50℃；浸提温度120℃，浸提时间3hr，固液比1：40。采用该工艺的粗多糖提取得率达14.88％，比热水浸提工艺的粗多糖得率提高了117.23％，比资料介绍的四次浸提法提高了75.06％。 本文系统地研究了羊栖菜粗多糖中蛋白质的分离方法，先后选用了Sevag法、酶法结合Sevag法、三氯乙酸法、1万分子截留量膜超滤、阴阳离子交换树脂法。发现用弱碱性阴和弱酸性阳离子交换树脂串联脱除粗多糖中蛋白质，效果理想，蛋白脱去除率可达91.76％，多糖回收率为92.56％，其中(多糖含量／蛋白含量)值高达42.27，经1万截留分子量透析膜透析，最后冷冻干燥后，得到淡黄色的羊栖菜多糖精粉，其中多糖含量达90.27％，蛋白质含量仅为浙江工业大学硕士学位论文羊栖菜多糖的提取和纯化研究3.巧%。该方法还起到部分脱色效果。 将脱蛋白后的多糖精粉用SePhadexG一200葡聚糖凝胶分离成大分子量多糖部分和小分子量多糖两部分。大分子量多糖部分用SePhacy1S一400共聚糖凝胶分离，得到一个分子量相当大的组分，经凝胶HPLC纯度鉴定为均一性物质，命名组分Fl，该组分含量占多糖精粉的28.45%;小分子量多糖部分进行SePhadexG一1 00葡聚糖凝胶柱层析，结果表明主要有三个组分fl、几、。，经凝胶HPLC标准分子量对照分析，三个组分分子量分别为8.2万、5.6万和1.4万;经计算，比例为:37.4%:15.1%:475%。 本实验研究有效地分离了羊栖菜粗多糖中蛋白质，制得羊栖菜多糖精粉，为羊栖菜在保健食品和药物研究和开发方面提供了良好的基础，同时为羊栖菜多糖提取和分离的工业化研究和生产提供了理论依据。更多还原

【Abstract】 Sargassum jusiforme(Harv.) Setchel is one of the plentiful brown seaweeds in east China sea. Its various biological activities (antitumoral, anticoagulant, antiviral, antioxdative, hypoglycemic and so on) have been reported in the literature. To make best use of the precious marine resources, the methods of extraction and purification of polysaccharide (SFP) from Sargassum fusiforme were studied in this paper.Three methods for extraction of SFP are discussed. The conditions of extracting SFP by boiling water, acid water and cellulase were investigated, result shows the optimum extraction temperature of enzymatic treatment is 50℃ optimum time is 3hr, optimum pH value is 3.0, add cellulase (16000U/g ) 8%, pectinase (20000U/g) 4%. The optimum extraction temperature of boiling water treatment is 102℃, optimum time is 3hr, weight of material: weight of water=l :40. Enzyme and high temperature extraction was combined to improve the low extraction rate of acid extraction and neutral boiling water extraction, and the yield could attain 14.88%. The best experiment factors are received Cellulase (16000U/g ) 8%, pectinase (20000U/g) 4%, pH3.0, weight of material : weight of water=1:40. The time of enzymolysis 3hr> the time of extraction 3hr the temperature of extraction 120℃. Compared with other extraction methods,enzymatic treatment made yield of polysaccharide increasing 117.23% and up to 75.06%, which was higher than the method of four time extractions mentioned in literaturesSeveral methods includeing, Sevag method, enzyme decomposition with Sevag method, trichloroacetic acid(TCA) sedimentation, ultrafiltration , anion and cation exchange resin chromatography were studied to remove the protein in the SFP. Based on experimental results and general consideration, it was suitable to adopt using anion and cation exchange resin in series chromatography once. After that, 91.76% protein of crude polysaccharides could be removed, the recovery rate of polysaccharide was 92.56%, and the ratio value of polysaccharides content to protein content was up to 42.27.Then after dialysis and freeze dryness, light yellow powder was produced. In this powder, polysaccharide and protein content was 90.27% and 3.15%.Higher molecular and lower molecular weight sections were gained by using Sephadex G-200 gel chromatograph. Then a polysaccharide fraction containing 1.5% protein was gained by Sephacryl S-400 chromatographing to the higher molecular. The purity of this fraction was proved by HPGPC and its molecular was more than 2×106 dalton and was named F1. The portion of F1 was 28.45% of fine polysaccharides. And the low molecular weight was chromatographed by Sephadex G-100 gel filtration, and three fraction f1, f2, f3 were gained , the molecular weight of them was 8.2×104(37.4%), 5.6×104 (15.1%), 1.4× 104(47.5%), the weight ratio was 38.2%, 16.1%, 45.7% separately.In this paper, purified powder of SFP was obtained by isolated protein in rude SFP effectively. The research offers some scientific basis for producing healthy foods and medicines. It provides theoretical foundation for industrialization of extraction and isolation polysaccharides from Sagasssum fusiforme.更多还原