CAJ | 학술논문

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。
목적：중시생산중대폐염극뢰백간균배양공예진행개진급우화。방법채용액체종합배양기대체반종합배양기재10 L화100 L중국려생물반응기중대폐염극뢰백간균진행배양，재10 L중국려생물반응기탐토불동적배양기배방、pH치、배양온도、교반전속、용양，공예삼수은정후，확대배양도100 L중국려생물반응기，병탐토배양과정중보가포도당적농도급보가방식등대세균농도급협막다당함량적영향。결과폐염극뢰백간균액체종합배양기가대체반종합배양기용우해균적배양，배양과정중유지pH치7．2、온도37℃、통기60 L/h、교반전속250 r/min、배양도2 h시개시이항속보가30 mL/L 40％포도당용액、배양시간위5 h，세균장세최호，수획적협막다당함량최고。결론폐염극뢰백간균적배양공예방대도100 L중국려생물반응기중，경과다차시험초보건립료은정적폐염극뢰백간균중시배양공예。
Objective To improve the cultural process and select the optimity for producing Klebsiella pneumoniae vaccine . Methods The integrated liquid medium was instead of semi-integrated medium for inoculating Klebsiella pneumoniae in bioreactor (10 L and 100 L of volume), and the effects of various factors (such as medium formulation, pH, culture tem-perature, stirring speed, oxygen concentration , supplemented glucose content and its supplementing time pon ) on the growth and polysaccharide yield were detected in bioreactor .Results The results showed that the integrated liquid medium can be used for inoculating Klebsiella pneumoniae to substitute for semi-synthetic medium .The optimum culture conditions are as follows:pH 7.2, temperature at 37℃, stirring speed at 250 r/m, 60 L/h ventilation, and supplementing 3 000 mL of 40%glucose solution into 100 L medium after inoculating two hours , and then continuously cultured for three hours , by this time the growth state of the bacteria is the best , polysaccharrde content is the highest .Conclusion Stable Klebsiella pneumonia fermentation for producing vaccine has been established by several validated test with 100 L bioreactor .