摘要：医用级透明质酸的制取有一定的困难，要求其分子量在180Da万以上，杂蛋白的含量低于0.1%。目前在除蛋白方面大部分还是在用活性炭处理。本课题以透明质酸发酵液为材料，研究活性炭对发酵液中的蛋白质，色素等一些杂质吸附作用。在实际纯化过程中，既要保证较高的蛋白去除率，又要保证透明质酸分子量和含量损失较小。因此选择合适种类的活性炭将有效提高实验效果，同时找出合适的吸附时间，温度，pH等吸附条件也至关重要。在本次研究中，设计了多组单因素实验，考察了活性炭种类、用量、吸附时间、温度、pH五个因素对除蛋白效果的影响情况。研究结果表明，在37℃下，调节pH为3.0，以6倍透明质酸质量的药用活性炭处理0.5小时所得实验效果最佳。22196
毕业论文关键词：透明质酸；分子量；蛋白去除率；活性炭
Protein separation during the extraction of medical hyaluronic acid
Abstract: There is certain difficulty in the production of medical hyaluronic acid with lower than 0.1% protein contented, because its molecular weight should be over 1.8 million Da. At present, activated carbon treatment is the most widely used in the process of removing protein. In this study, the hyaluronic acid broth from fermentation is used as the material to discover acticarbon’s adsorptive capacity of pigments and other impurities in the fermentation broth. The high protein removal rate should be ensured while the loss of both molecular weight and hyaluronic acid content should be slight. So the appropriate kind of acticarbon will efficiently improve the result, meanwhile the suitable adsorption time, temperature, pH and other adsorption conditions will also play important roles in the result. In this study, multiple groups of single factor experiment have be designed to study the effects of activated carbon types, dosage, adsorption time, temperature and pH on removing protein. The results show that, under the temperature of 37 ℃, when the pH was adjusted to 3.0, 6 times the hyaluronic acid quality of medicinal charcoal treating half an hour will produce the best result.
Keywords: Hyaluronic acid; molecular weight; protein removal rate; acticarbon
目 录
1 绪论 1
1.1 透明质酸综述 1
1.2 透明质酸的分布简述 1
1.3 透明质酸的理化性质简述 1
1.4 透明质酸的生理功能简述 2
1.4.1 透明质酸的保湿性能 2
1.4.2 透明质酸参与构成蛋白糖聚糖聚合体 2
1.4.3 透明质酸对细胞生物学的影响 3
1.4.4 透明质酸对免疫细胞的作用 3
1.4.5 对创伤愈合的影响 3
1.5 透明质酸提取工艺研究现状 4
1.5.1 过滤法 4
1.5.2 凝胶层析法 4
1.5.3 离子交换层析法 5
2 本课题的研究背景和目的 5
2.1课题背景 5
2.2 课题目的 6
3 药品与仪器 6
3.L试剂与仪器设备 6
3.2 方法与技术 7
3.2.1 试剂配制 7
3.2.2 蛋白质浓度标准曲线的制作及样品测定 7
3.2.3 葡萄糖醛酸标准曲线的制作及样品测定 8
3.2.4 相对分子量的测定 9
4 试验方法 9
4.1不同种类的活性炭对除蛋白的影响 9
4.2 活性炭的用量对除蛋白的影响 10
4.3 活性炭处理时的温度对除蛋白的影响 10
4.4 PH对除蛋白的影响 11 医用级透明质酸提取过程中杂蛋白的去除:http://www.youerw.com/shengwu/lunwen_14712.html