海洋鱼类弧菌病疫苗的制备--新型简易鳗弧菌(Vibrio anguillarum)培养基优化①

以一株从我国山东海域爆发弧菌病鲈鱼体内分离得到的病原菌鳗弧菌（Vibrio anguillarum MV425 O1型）为研究对象，对其实现高密度生产海洋鱼类弧菌病疫苗的培养基进行了优化研究。研究表明，碳源的NaCl造成的渗透压以及由发酵碳源和氮源导致的pH值变化是影响细胞正常生长的重要因素。海水中微量元素（除铁元素）对菌体生长的作用可被酵母浸出物补偿，高铁浓度有助于菌体生长。在确定显著影响因子的基础上，采用可旋转中心复合设计方案和响应面法进行了统计模型优化，优化设计有效地提高了细胞培养密度，实现在摇瓶条件下细胞干重5．8g/L和30L规模反应器中26．2g/L的较好的初步放大结果，并可替代海水培养基。

Production of Marine Fish Vaccine for Vibriosis from Vibrio anguillarum: Optimization of New Culture Medium for High-density Culture

A freshwater culture medium for high density culture was optimized for the production of marine fish vaccine from a marine fish pathogen Vibrio anguillarum MVM425 at shake flask level. In this study, it was found that cell growth inhibition by high substrate concentrations was more due to high osmotic stress of the culture environment, produced by carbon sources and NaCl, and low pH caused by fermentation of carbon and nitrogen sources. The effect of trace elements in seawater (except for iron) on cell growth could be compensated by yeast extract. Increased cell growth was achieved by high iron concentration. Central composite rotatable design and response surface methodology were employed to derive a statistical model for media optimization towards high density culture. This statistical design was very effective in improving the cell density up to 5.8 g dry weight per liter with the batch culture in shake flask. Maximum cell dry weight 26.2g/L was achieved at 30L bioreactor using optimized media.