兽疫链球菌NW-162发酵产生的透明质酸分离纯化研究

Separation and purification of hyaluronic acid by fermentation of Streptococcus zooepidemicus NW-162

对兽疫链球菌诱变株NW-162发酵液中透明质酸(HA)的分离提取工艺方法进行了对比研究。采用乙醇沉淀分离透明质酸、链霉蛋白酶酶解结合氯仿除去蛋白质及经氯代十六烷吡啶分级分离工艺,结果表明:乙醇与上清液的体积比为2.5∶1.0时沉淀透明质酸25 h最为经济;酶解结合氯仿去除蛋白质较氯仿单独作用时,透明质酸产量及纯度均有一定提高,再经氯代十六烷吡啶分级分离后,可得纯度为71.5%、蛋白质质量分数2.8%、产量0.747 g/L、相对分子质量725×103的透明质酸产品。

The comparative study on the process of separation and extraction of hyaluronic acid （HA） by fermentation of Streptococcus zooepidemicus mutant NW-162 was carried out. The processes for precipitation by ethanol, zymolysis combined with deproteinization by chloroform and fractional separation by cetylpyridinium chloride （CPC） were adopted. The results indicate that precipitating HA for 25 h with the volume ratio of ethanol to supernatant 2.5 ： 1.0 is the optimal. Compared with that of action by chloroform solely, the technology of zymolysis combined with deproteinization by chloroform improves the production and purity of HA in a certain extent. After fractional separation with CPC, the final product of HA with purity 71.5%, protein mass fraction 2.8%, output 0.747 g/L, relative molecular mass 725 × 10^3 was obtained.