细菌硫酸软骨素发酵生物技术研究
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
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