重组大肠杆菌DE-pET-P46的培养基优化和高密度发酵

Medium Optimization and High Density Fermentation for the Recombinant E. coli DE-pET-P46

关键词:  猪肺炎支原体  P46蛋白  高密度发酵  响应面

Key words:  Mycoplasma hyopneumoniae  P46 protein  high cell density cultivation  response surface analysis

摘要:

为提高重组蛋白的生产效率，利用响应面试验设计技术，对表达猪肺炎支原体P46蛋白的重组大肠杆菌DE-pET-P46的培养基进行了优化，确定了培养基各营养成分及其最佳配比。在相同培养条件下，优化培养基比LB培养基的菌体浓度高出一倍。在生物反应器中利用优化培养基发酵培养该重组大肠杆菌，培养物的湿菌重达39.5 g/L。SDS-PAGE电泳结果显示，高密度发酵对重组大肠杆菌DE-pET-P46的rP46的表达和占细胞总蛋白比例均没有明显的改变。

Abstract:

For higher efficiency and lower cost of the protein production, response surface analysis(RSA) was used for culture optimization of the recombinant E. coli which could express Mycoplasma hyopneumoniae P46 protein, and the best ratio of each medium component was determined. Under the same culture conditions, the cell concentration of the optimized culture is twice higher than LB medium. The medium used in a bioreactor for the recombinant E. coli DE-pET-P46 culture fermentation, the wet bacteria weight of the culture is up to 39.5 g/L. SDS-PAGE electrophoresis showed that there is no obvious difference on the rP46 expression of the recombinant E. coli DE-pET-P46 between cultivated with bioreactor and with shaking incubator.