微生物发酵透明质酸的制备及测定  

Study on the Preparation and Mensuration of Hyaluronic Acid from Fermentation Broth

作　　者：吕磊[1] 吕炜锋[1] 姚文兵[1] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009

出　　处：《药物生物技术》2009年第2期162-165,共4页Pharmaceutical Biotechnology

摘　　要：为提高透明质酸成品质量和收率,针对透明质酸发酵液中菌体和蛋白质的物化性质,用乙醇对发酵液进行沉淀得透明质酸粗品,粗品溶解后用硅藻土、活性炭吸附,再经滤板过滤,然后加入含氯代十六烷基吡啶的氯化钠溶液沉淀,溶解,乙醇沉淀等步骤后溶液经滤膜过滤,醇沉,干燥。所得透明质酸为白色粉末,其回收率约为79%,蛋白质含量低于0.1%,相对分子质量达180万,活性炭除内毒素后,内毒素限值符合欧洲药典标准(<0.5Eu/mg)。To improve the quality and yield of HA, a novel pretreatment was developed aiming at removing cells and proteins in fermentation broth. Crude HA was obtained by precipitating using ethanol. HA was then dissolved in water and was filtrated with the mixture of kieselguhr and active carbon. NaCl solution containing cetylpyridinium chloride（CPC） was added into HA solution, following which the cetylpyridinium salt precipite was obtained and was then re-dissolved again in water. The process of precipitation by ethanol and re-dissolution were repeated. Final fine product was gained after film filtration, ethanol precipitation and drying. The yield of the final product was 79 % and the colour was white, with protein content lower than 0. 1%. Its relative molecular mass was as high as 1.8× 10^6. When the endotoxin was removed by active carbon, the bacterial endotoxin has reached medical grade, coming up to the standard of European Pharmacopoeia （〈0. 5Eu/mg）.

关 键 词：透明质酸 医药级 分离纯化 氯代十六烷基吡啶 

分 类 号：TQ460.6[化学工程—制药化工]