马疫链球菌IMF401发酵生产透明质酸的提取工艺研究
本文章对马疫链球菌IMF401为出发菌发酵生产的透明质酸发酵液进行提取条件的优化，采用膜抽滤与活性炭吸附相结合的方式，对发酵液进行预处理和后期提取纯化。通过单因素实验与正交优化实验，最终确定发酵液预处理条件为：在70℃条件下，以型号SH的滤纸板作为抽滤介质，以硅藻土SYS为助滤剂（40g/L）进行抽滤；后期分离纯化条件为：在30℃温度下，加入C:HA（质量比）为2:1的活性炭，采用磁力搅拌或者摇床摇匀处理4h，后抽滤得滤液，滤液3倍乙醇醇沉，取沉淀真空干燥得到成品。成品透明质酸钠经检测醛酸含量为41.78%，分子量约1.63×106Da，蛋白质含量为1.30‰；含水量约10.28%。
毕业论文关键词：马疫链球菌；透明质酸；工艺优化；预处理；分离纯化
Study on extraction process of fermentative production hyaluronic acid from Streptococcus equi IMF401
ABSTRCT: Hyaluronic acid is one king of acidic mucopolysaccharido，hyaluronic acid, is made up with N- acetyl glucosamine and glucuronic acid by β-l, 4 glycosidic linkages and β-l, 3-glycosidic bond alternately connected together Acid mucopolysaccharides. Hyaluronic acid is widely distributed in the extracellular matrix, connective tissues and organs of higher animals.. With special pHysiological efficiency, and in the medical, cosmetic and health products and other fields can be widely used [1,2].
The present article of Streptococcus equi IMF401 of fermentation production of hyaluronic acid fermentation liquid for optimization of extraction conditions, combined with membrane filtration and activated carbon adsorption, of fermentation liquid of pre - and post - extraction purification. Through single factor experiments and orthogonal optimization experiments, and ultimately determine the fermentation liquid pre treatment conditions: under the condition of 70 DEG C, filter board with types of SH as the filtration medium, diatomite sys as filter aid (40g/L) filtration; anapHase separation purification conditions: under the temperature of 30 DEG C, adding C:HA (quality ratio) is 2:1 activated carbon using magnetic stirring or shaking shaking treatment for 4h, after filtration to obtain filtrate, filtrate 3 times of ethanol alcohol precipitation, precipitation vacuum drying to obtain the finished product. The content of sodium hyaluronate was 86.5%, and the molecular weight of sodium hyaluronate was 1.63×106 Da，Protein content was 1.30 ‰，and the content of water was about 10.28%。
Keywords: Streptococcus equi; hyaluronic acid; process optimization; pretreatment; separation and purification
目录
第一章 前言 6
1.1 透明质酸的结构、分布和性质 6
1.1.1 透明质酸的结构 6
1.1.2 透明质酸的分布 6
1.1.3 透明质酸的性质 6
1.2 透明质酸的用途 7
1.2.1 透明质酸在化妆品工业的应用 7
1.2.2 透明质酸的临床应用 8
1.2.3 在保健品方面的应用 8
1.3发酵液中透明质酸的分离和提纯 9
1.3.1 微生物发酵法 9
1.3.2 动物组织提取法 9
1.3.3 发酵液中HA的提取 10
1.4 本课题的研究目的和意义 10
第二章 透明质酸发酵液预处理条件优化 12