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目的:为Ompk亚单位疫苗生产提供参数。方法:利用振荡和发酵培养,测定不同培养时间的菌液浊度和蛋白诱导表达效果。结果:工程菌30℃摇瓶培养10h,种子罐培养8～10h,较为合适;摇瓶和5L发酵罐诱导表达培养,升温42℃诱导表达,7h效果较佳;经5批次50L、500L发酵罐诱导表达培养,均可得到重组蛋白表达的工程菌体,50L和500L发酵罐最高生物量分别为5.32g/L和6.38g/L,平均可达到3.75 g/L和4.74 g/L;适当延迟升温诱导前的培养时间,可提高工程菌的得率。结论:初步确定了重组蛋白工程菌规模化诱导表达培养方法,利用500L发酵罐培养可得到诱导表达的工程菌体。
Objective: To provide parameters for Ompk subunit vaccine production. Methods: Osmosis and fermentation were used to determine the turbidity and the protein expression induced by different culture time. Results: The engineered bacteria were cultured in shake flask at 30 ℃ for 10h, and the seed tank was cultured for 8-10h, which was more suitable. The shake flask and 5L fermenter were induced to express at 42 ℃ for 7h, The maximum biomass of 50L and 500L fermentors was 5.32g / L and 6.38g / L, respectively, with an average of 3.75 g / L and 4.74 g / L, respectively. Appropriate Delayed induction of culture temperature before induction, can improve the yield of engineering bacteria. Conclusion: The method of scale-induced expression of recombinant protein engineering bacteria has been preliminarily determined, and the engineered bacteria that can be induced to express in a 500L fermenter can be obtained.