溶氧探测补料技术发酵生产类人胶原蛋白的研究  

Fed-batch production of human-like collagen with recombinant Escherichia coli using feed-up DO-transient control

作　　者：薛文娇[1] 范代娣[2] 

机构地区：[1]陕西省微生物研究所,陕西西安710043 [2]西北大学化工学院陕西省可降解生物医药材料重点实验室,陕西西安710069

出　　处：《化学工程》2011年第10期6-10,共5页Chemical Engineering(China)

基　　金：国家自然科学基金资助项目(31000019)

摘　　要：在重组大肠杆菌发酵生产类人胶原蛋白(HLC)过程中,快速确定不同反应器中的补料速率,抑制乙酸产生,高水平表达HLC。采用溶氧探测补料技术,在不同规模反应器中,分批-补料培养重组大肠杆菌生产HLC,根据脉冲补料方式时溶氧信号的响应特征来辩识是否产生乙酸并确定补料速率。在实验室规模获得的最终细胞密度和HLC质量浓度分别为69.1,13.1 g/L,该值和以前根据比生长速率优化的结果基本一致;放大到中试规模(500 L)时,非诱导培养时最终细胞密度由实验室规模的80.3 g/L下降到54.1 g/L,但乙酸质量浓度和实验室规模基本相同;优化诱导时机后获得的最终细胞密度和HLC质量浓度分别为51.7,9.6 g/L,表达水平和实验室规模基本相同。利用溶氧探测补料技术分批-补料培养重组大肠杆菌,可以抑制乙酸的产生,高水平表达HLC。To control the acetate production and achieve a high-level expression of the target protein in the different-scale bioreactors during fed-batch production of human-like collagen（HLC） with recombinant Escherichia coli.A probing feeding strategy using feed-up DO-transient control was employed in two different-scale bioreactors and the dissolved oxygen response signal was used to detect the acetate production in pulsed fed-batch cultivation of E.coli.The final dry cell weight（DCW） of 69.1 g/L and HLC mass concentration of 13.1 g/L were obtained on the laboratory scale and the results are similar to that optimized in the previous study.When the process was scaled up to pilot-scale（500 L）,the final DCW decreased from 80.3 g/L to 54.1 g/L with non-induced cultivations while acetate mass concentration was similar on the both scales.The difference in DCW was mainly resulted from the difference in oxygen transfer capacity.The final DCW of 51.7 g/L and HLC mass concentration of 9.6 g/L were obtained on the pilot-scale upon optimization of induction timing,and the results are satisfied based on the HLC content.The feed-up DO-transient control strategy can be successfully employed to control the acetate production and achieve a high-level expression of the target protein during the fed-batch production of HLC with recombinant E.coli.

关 键 词：溶氧探测补料技术 乙酸 重组大肠杆菌 类人胶原蛋白 

分 类 号：TQ936[化学工程]