由透明质酸发酵液制备高纯度透明质酸的方法  

The Method of Preparation of Highly Purified Hyaluronic Acid from Fermentation Broth

作　　者：郭育涛[1] 江元汝[1] 陈斌[1] 陈双莉[1] 

机构地区：[1]西安建筑科技大学理学院化学系,西安710055

出　　处：《天然产物研究与开发》2008年第4期725-727,747,共4页Natural Product Research and Development

基　　金：陕西省教育厅专项基金(2001JK210)

摘　　要：为简化由透明质酸发酵液制备高纯度透明质酸的过程,本实验利用氯苯除去菌体,由乙醇分离得到透明质酸提取物,向提取物中加入由4.2 g/L碳酸氢钠和5.3 g/L碳酸钠以体积比为45∶5组成的水溶液及胰蛋白酶进行水解。水解后所得分离物在磷酸二氢钠含量为0.45 g/L,磷酸氢二钠含量为0.05 g/L及氯化钠含量为11.6 g/L的水溶液中溶解,向其中加入3倍体积的95%乙醇,得到沉淀,绝干的沉淀物中透明质酸质量比为98.9%。随后,经活性炭以及透析处理。最终透明质酸质量比为99.996%,相对分子量5.8×105。总回收率96.17%。In order to simplify processes of preparation of purification of hyaluronie acid （HA） from fermentation broth, in this study a lot of bacterial cells were removed via ehlorbenzene, and extracts including HA was separated by ethyl alcohol. Then, the extracts and trypsin were added to a basic water solution which was composed of sodium bicarbonate solution of mass concentration 4.2 g/L and sodium carbonate solution of mass concentration 5.3 g/L according to volume ratio 45： 5,and protein in the extracts was hydrolyzed. Hydrolysates separated by ethyl alcohol,including HA,were dissolved in saliniferous water solution in which prelate salt content reach 0.45 g/L and sodium biphosphate content reach 0.05 g/L and sodium chloride content reach 11.6 g/L. When 3 times amount of 95% ethyl alcohol was added to the solution, precipitates including HA was got. Mass ratio of HA in oven-dried precipitates was up to 98.9%. Then, charcoal was added to the precipitates dissolved in the saliniferous water solution and the solution was dialyzed for removal of pyrogen. Lastly,mass ratio of HA in purified sample was up to 99.996% ;relative molecular weight of HA was 5.8 × 10^5 ; total recovery of HA was 96.17%.

关 键 词：透明质酸 纯化 胰蛋白酶 发酵液 

分 类 号：Q530[生物学—生物化学] R284.3[医药卫生—中药学]