b型流感嗜血杆菌荚膜多糖发酵培养基的优化

Optimization of Media for Capsular Polysaccharide Fermentation of Haemophilus influenza Type b

探明b型流感嗜血杆菌荚膜多糖发酵的最适培养基及接种量,为降低其疫苗荚膜多糖制备成本提供理论依据,设计6种培养基进行摇瓶培养,检测发酵上清液中的菌体量,筛选最适培养基,并在此基础上优化接种浓度及氯化血红素、辅酶I添加量进行发酵罐培养,测定荚膜多糖含量,评估培养效果。结果表明:接种后OD600值为0.1的接种量较为适宜b型流感嗜血杆菌生长;3号改良培养基(选择性大豆蛋胨、酵母浸粉、酸水解酪蛋白、K2HPO4、氯化钠、葡萄糖、氯化血红素、辅酶I分别为1%、1%、0.50%、0.25%、0.50%、0.70%、25mg/L和20mg/L)对b型流感嗜血杆菌液体进行发酵罐发酵可获得大量菌体且稳定,荚膜多糖含量为(105.6±13.2)mg/L。

The optimal medium was selected from six media by detecting the bacteria quantity in the fermented supernatant after shake-flask culture and the capsular polysaccharide content of Haemophilus influenzatype b（Hib）cultured in the optimal medium with different inoculation quantity and different chlorhematin and coenzymeⅠ concentration was detected to explore the optimal medium and inoculation quantity for capsular polysaccharide fermentation of Hib and provide the theoretical basis for reducing the preparation cost of capsular polysaccharide of Hib.Result：The inoculation quantity for Hib growth is0.1 OD600 value after inoculation.The capsular polysaccharide content of Hib cultured in the improved medium including 1%selectivity soy peptone,1%yeast extract powder,0.50%acid hydrolyzed casein.0.25% K2 HPO4,0.50% NaCl,0.70% glucose,25 mg/L chlorhematin and 20 mg/L coenzymeⅠcan reach（105.6±13.2）mg/L.