-
摘要：硫酸软骨素一般存在动物的软骨组织中，是一类糖胺聚糖。由多种酸性多糖的混合，其多糖是通过共价键与蛋白质相连形成蛋白聚糖。硫酸软骨素具有预防心血管、提高免疫力、调节脂类代谢、抗衰老等功能。在生物医学、食品等领域中被广泛应用。本毕业论文主要研究硫酸软骨素的提取工艺。原料采用粉碎后的猪软骨，通过蛋白酶处理，再进行提取和纯化。用硫酸-咔唑法分析方法确定硫酸软骨素的提取率，以提取率为考察指标，通过正交试验优化提取工艺条件，试验结果表明，最佳的提取工艺条件为：酶解时间26h、酶浓度120U/mL、料液比（w/v）1:15、提取温度55℃。正交试验分析表明，酶解时间对结果影响最大，酶浓度对结果的影响次之，而料液比和提取温度对结果产生的影响相对小些。粗提取物再经过离子交换柱分离纯化，离子交换柱采用Amberlite IRA-933 型阴离子交换树脂。结果显示Amberlite IRA-933 型柱能选择吸附对硫酸软骨素，洗脱时，使用氯化钠作为洗脱液，获得较好的纯化效果。纯化的硫酸软骨素样品外观为白色粉末，略带咸。21196
- 上一篇：几种多肉植物水培条件下抗盐实验
- 下一篇：地被菊耐涝性研究+文献综述
关键词：硫酸软骨素；酶解；提取
Study on the extraction of chondroitin sulfate
Abstract： Chondroitin sulfate, generally present in animal cartilage, is a mix of some acidic polysaccharides. It belongs to proteoglycan glycosaminoglycans, which is connected with protein via covalent bond. Chondroitin sulfate can prevent disease of cardiovascular, improve immunity, regulate lipid metabolism, anti-aging and others. So it has been widely used in the biomedical, food and other fields. In this paper, we use pig cartilage as raw material and enzymatic hydrolysis as extraction process. Then sulfuric acid-carbazole assay was conducted to assess extraction rate. Finally, we choose the optimum extraction technology programs and best purification assay. The results showed that, by orthogonal to the rate of chondroitin sulfate extraction, hydrolysis time had greatest impact on the extraction rate, followed by enzyme concentration. The ratio of solid to liquid and extraction temperature on the results was considered to have little impact. The best extraction conditions were : hydrolysis time 26h, enzyme concentration 120U/mL, solid-liquid ratio (w / v) 1:15, extraction temperature 55 ℃. We also found that Amberlite IRA-933 anion exchange resin column had strong capability for eluting chondroitin sulfate. And sodium chloride was a good eluent with high purification rate. Finally, we got purified chondroitin sulfate as white, odorless and salty powder.
Key Words：Chondroitin sulfate; enzyme; extract
目录
1 绪论 1
1.1硫酸软骨素概述 1
1.2硫酸软骨素的作用 1
1.3 硫酸软骨素的提取工艺方法 2
1.4课题研究目的和意义 2
1.5 课题研究的内容 3
2 实验材料和方法 3
2.1 实验试剂及仪器 3
2.2 实验方法 4
2.2.1 蛋白酶活力的测定 4
2.2.3 硫酸软骨素的不同工艺条件对提取的影响 5
2.2.3 蛋白酶最优反应条件的确定 5
2.2.4 Amberlite IRA-933 型阴离子交换树脂柱纯化 5
2.2.5 旋转蒸发浓缩 6
2.3分析方法 7
2.3.1已糖醛酸法测定硫酸软骨素含量 7
3 实验结果与讨论 8
3.1 蛋白酶活力的测定 8
3.2 胰蛋白酶酶解工艺条件的确定 9
3.2.1 预实验 9
3.2.2 酶浓度对提取效果的影响 10
3.2.3 原料与提取液之比对提取效果的影响 11