摘要：硫酸软骨素是是一类硫酸化链状黏多糖[1]，硫酸软骨素是由D-葡萄糖醛酸和N-乙酰-D-氨基半乳糖以β-1,3-糖苷键连接形成二糖，而二糖单位之间以β-1,4-糖苷键连接而成，是一种高分子化合物[2]。本文进行了微生物发酵法合成硫酸软骨素的研究，建立起了发酵液中的硫酸软骨素的提取、测定以及分析的比较系统的一套方案。微生物发酵液通过一定的预处理操作及醇沉操作后，采用碱法-酶法结合醇沉法对其中的硫酸软骨素进行提取，经过阳离子交换树脂分离纯化，用高效液相色谱法进行含量测定以及单糖组分分析，采用葡聚糖凝胶色谱法测定相对分子质量，同时根据盐酸水解-硫酸钡重量法对其中的硫进行定量分析。研究发现，对发酵液中提取的硫酸软骨素中进行分离纯化后，其纯度可达65%-70%，其中主要含有的单糖是葡萄糖，相对分子质量为36602，硫含量为4.36%.30841
毕业论文关键词：硫酸软骨素；微生物发酵法；多糖组分分析；葡聚糖凝胶色谱；
Study on the fermentation technology of bacterial chondroitin sulfate
Abstract: Chondroitin sulfate is a kind of sulfated chain mucopolysacchride, is mainly consisted of  D - glucuronic acid and N-acetyl galactosamine to beta 1,3 glucosidic bond connected to form a disaccharide, disaccharide units in beta 1,4 glycosidic bond connected, is a kind of high molecular compound. In this paper, it does some research of microbial fermentation method for the synthesis of chondroitin sulfate, and then establish the fermentation liquid of chondroitin sulfate extraction, determination and analysis system of a scheme. After some pre-processing and alcohol precipitation operation, through the method of alkali and enzyme hydrolyzing combined with alcohol precipitation, the chondroitin sulfate can be extracted from the broth, and then separating and purifying by cation exchange resin, using high performance liquid chromatography for content determination and monosaccharide composition analysis, using Sephadex chromatography for the determination of relative molecular mass, at the same time, according to the quantitative analysis of the sulfur hydrochloric acid hydrolysis - barium sulfate gravimetric method. It found that the purity of chondroitin sulfate extracted from fermentation liquid,can reach 65% - 70%, which mainly contain the monosaccharides glucose, relative molecular mass for 36602 ,and the sulfur content is about 4.36%.
Key words:  Chondroitin sulfate； microorganism fermentation；polysaccharide composition analysis； Sephadex chromatography
目  录
1 前言    1
 1.1 硫酸软骨素的组成、分布、结构和生化特性    1
 1.2 硫酸软骨素的药用功效    2
 1.3 硫酸软骨素的分离纯化    2
  1.3.1 阳离子交换树脂法    2
  1.3.2 高效液相色谱法    3
  1.3.3 葡聚糖凝胶色谱法    3
 1.4 硫酸软骨素的含量测定    4
  1.4.1 多糖含量测定    4
  1.4.2 硫含量测定    4
 1.5 本课题主要研究内容与研究意义    5
  1.5.1 研究内容    5
  1.5.2 研究意义    6
2 材料与方法    8
 2.1 材料与仪器设备    8
  2.1.1 菌种    8
  2.1.2 实验材料    8
  2.1.3 实验仪器与设备    9
  2.1.4 培养基及试剂的制备    10
 2.2 实验方法    11
  2.2.1 种子制备及微生物发酵    11
  2.2.2 正交试验优化种子和发酵条件    11 细菌硫酸软骨素发酵生物技术研究:http://www.751com.cn/shengwu/lunwen_26800.html