摘要
为提高重组质粒的生产效率,利用响应面分析中的Plackett-Burman试验设计、最陡爬坡试验和Box-Behnken试验,对影响该工程菌质粒表达的培养基组分进行筛选和优化,以确定该工程菌的最佳培养基组分。结果显示:蔗糖、硫酸铵分别为最佳碳源、无机氮源;酵母浸粉、蔗糖和(NH4)2SO4最佳质量浓度分别为7.50,17.50,7.50g/L。在相同培养条件下,优化后培养基质粒产量是LB培养基质粒产量2倍。使用100L反应器高密度培养时利用优化的培养基发酵培养重组大肠杆菌,质粒质量浓度达到115.25mg/L,比优化前提高88.52%,质粒各项指标稳定,免疫原性优良。此试验数据为规模化生产提供参考数据。
For higher efficiency and lower cost of the plasmid production, Plackett-Burman Design and Box-Behnken Design in response surface methodology were used for culture optimization of the recombinant E. coli DH5α-pCAGGoptiHA5 which could affect the engineered bacteria plasraids expression. The result shows that sucrose was the best carbon source, （NH4）2SO4 was the best inorganic nitrogen source. The optimum concentration of yeast extract,sucrose, （NH4）2SO4 was 7.50,17.50 and 7.50 g/L respectively. Compared with LB medium, the plasmid concentraton increased 100% with the same culture condition. When amplified to 100L-scale bioreactor of high density fermentation with the optimum medium,the concentration of recombinant E. coll. plasmid increased to 115.25mg/L,88.52% higher than that original culture,The plasmid had stable characteristics and good immunogenicity. The results would be useful for large-scale fermentation.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第11期2076-2082,共7页
Chinese Journal of Veterinary Science
基金
国家国际合作专项(2014DFA31180)
国家公益性行业(农业)科研专项(31372463)
关键词
重组质粒
高密度培养
质粒表达
规模化生产
recombinant plasmid
high density fermentation
plasmids expression
large-scale production
