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目的中试生产中对肺炎克雷伯杆菌培养工艺进行改进及优化。方法采用液体综合培养基代替半综合培养基在10 L和100 L中国丽生物反应器中对肺炎克雷伯杆菌进行培养,在10 L中国丽生物反应器探讨不同的培养基配方、pH值、培养温度、搅拌转速、溶氧,工艺参数稳定后,扩大培养到100 L中国丽生物反应器,并探讨培养过程中补加葡萄糖的浓度及补加方式等对细菌浓度及荚膜多糖含量的影响。结果肺炎克雷伯杆菌液体综合培养基可代替半综合培养基用于该菌的培养,培养过程中维持pH值7.2、温度37℃、通气60 L/h、搅拌转速250 r/min、培养到2 h时开始以恒速补加30 mL/L 40%葡萄糖溶液、培养时间为5 h,细菌长势最好,收获的荚膜多糖含量最高。结论肺炎克雷伯杆菌的培养工艺放大到100 L中国丽生物反应器中,经过多次试验初步建立了稳定的肺炎克雷伯杆菌中试培养工艺。
Objective Pilot production of Klebsiella pneumoniae culture process to improve and optimize. Methods Culture medium for Klebsiella pneumoniae was cultured in 10 L and 100 L Chinese bioreactors using liquid synthetic medium instead of semi-synthetic medium. Different culture medium formulations, pH value, Culture temperature, stirring speed, dissolved oxygen, and the process parameters were stable, the culture was expanded to 100 L bioreactor in China and the effect of adding glucose and supplementation on the bacterial concentration and capsular polysaccharide content . Results The Klebsiella pneumoniae liquid composite medium could be used in the culture of the strain instead of semi-synthetic medium. The culture process was maintained at pH 7.2, temperature 37 ℃, aeration 60 L / h, stirring speed 250 r / min, At 2 h, 30 mL / L of 40% glucose solution was added at a constant speed for 5 h. The bacteria grew best and the content of capsular polysaccharide was the highest. Conclusion The culture process of Klebsiella pneumoniae was amplified to 100 L in China Bioreactor. After several experiments, a stable pilot culture process of Klebsiella pneumoniae was established.