鲨鱼软骨中硫酸软骨素的提取工艺研究

论文编号:ZY095    论文字数:11581,页数:22

摘要：以鲨鱼软骨为原料，采用碱液提取，中性蛋白酶水解，三氯乙酸沉淀蛋白酶，乙醇沉淀粘多糖的工艺,提取了硫酸软骨素。研究了水解鲨鱼软骨的工艺参数，通过正交实验获得中性蛋白酶水解鲨鱼软骨提取硫酸软骨素的最优条件，结果表明，从鲨鱼软骨中提取硫酸软骨素的最佳工艺条件为：用质量浓度2%的NaOH溶液60oC下提取6h．再用质量浓度2%的NaOH溶液提取3h，提取液加人鲨鱼软骨量质量分数1.0%的中性蛋白酶，40-50oC水解4h，用三氯乙酸沉淀蛋白质，加入4-5倍体积的体积浓度96%的乙醇沉淀，在最佳条件下平均提取率为20.1%。产品为白色粉末。

关键词：粘多糖；硫酸软骨素 ；提取；纯化

Abstract：In this paper, the Shark cartilage was chosed as the raw material. The technology of the extraction with sodium hydroxide, the hydrolization with dispase, the precipitation of proteinum with trichloroacetic acid and the precipitation of mucoitin with ethanol were mainly studied. The optima conditions of the extraction of chondroitin sulfate were determined by orthogonal experiments  in this paper. The result shows that the most suitable and proper conditions of the extraction of chondroitin sulfate from Shark cartilage were as following : soaked with 2%NaOH for 6 hours at 60oC first, then soaked with 2%NaOH of the first for 3 hours. And then 1.0% dispase were added to the extracts for hydrolization at 40-50oC for 4 hours. The proteinum was precipitated with trichloroacetic acid, and then 4-5 times volume of 96% ethanol were added for the precipitation of mucoitin. The average extraction ratio was 32.2％,and the product was white powder.

Keyword：mucoitin;  chondroitin sulfate;  extraction;  purification

目  录
中文摘要………………………………………………………………………………Ⅰ
英文摘要……………………………………………………………………………Ⅱ
目录…………………………………………………………………………………Ⅲ
1绪论………………………………………………………………………………1
 1.1引言……………………………………………………………………1
 1.2 硫酸软骨素的结构与性质………………………………………………2
      1.2.1 结构……………………………………………………………………2
      1.2.2理化性质………………………………………………………………2
 1.3 鲨软骨粘多糖的应用……………………………………………………2
 1.4 提取路线分析……………………………………………………………3
  1.4.1 酶水解法……………………………………………………………3
  1.4.2 碱提取法……………………………………………………………4
  1.4.3 稀碱和酶结合法……………………………………………………4
 1.5 提纯路线分析………………………………………………………………5
  1.5.1 盐沉淀法……………………………………………………………5
  1.5.2 利用电离性质分离法………………………………………………5
  1.5.3 凝胶过滤色谱分离法………………………………………………5
  1.5.4 电泳分离法…………………………………………………………5
  1.5.5 超滤膜分离法………………………………………………………6
2实验部分…………………………………………………………………………7
 2.1 原料，试剂与仪器…………………………………………………………7
  2.1.1原料…………………………………………………………………7
  2.1.2试剂…………………………………………………………………7
  2.1.3仪器…………………………………………………………………7
    2.2 实验流程……………………………………………………………………8
 2.3 实验方案……………………………………………………………………8

 2.3.1 原料预处理…………………………………………………………8
 2.3.2 碱解…………………………………………………………………8
 2.3.3 酶解…………………………………………………………………8
 2.3.4 醇析…………………………………………………………………8
 2.3.5 精制…………………………………………………………………9
 2.3.6 产品检测……………………………………………………………9
3 结果与讨论…………………………………………………………………………10
 3.1 碱解因素对产物的影响……………………………………………………10
 3.2 酶解因素对实验结果的影响………………………………………………11
 3.2.1 酶用量的彭响………………………………………………………11
 3.2.2 酶解时间的影响……………………………………………………12
 3.2.3 温度的影响 ………………………………………………………12
 3.3 三氯乙酸对产物的影响……………………………………………………13
 3.4 醇析条件对产物的影响……………………………………………………14
 3.5 精制条件对产物的影响……………………………………………………14
 3.6 产品表征……………………………………………………………………14
4 总结与展望………………………………………………………………………16
致谢…………………………………………………………………………………17
参考文献……………………………………………………………………………18