重组大肠杆菌分批-补料培养生产类人胶原蛋白的过程控制  

A study on the process control for production of human-like collagen in fed-batch culture of escherichia coli BL 21

作　　者：骆艳娥[1] 范代娣[1] 花秀夫[1] 张兮[1] 王小刚[1] 

机构地区：[1]西北大学化工学院,陕西西安710069

出　　处：《西北大学学报（自然科学版）》2005年第2期180-183,共4页Journal of Northwest University（Natural Science Edition）

基　　金：国家科技攻关重大专项基金资助项目(2003DA901A32);国家自然科学基金资助项目(20476085)

摘　　要：目的优化重组大肠杆菌高密度发酵的调控工艺。方法通过考察比生长速率、3种控氧方式以及诱导强度对细胞和类人胶原蛋白产量的影响,确定最佳的发酵工艺。结果比生长速率显著影响类人胶原蛋白的产量,且最适比生长速率为0.15～0.20h-1;在提高罐压的条件下,细胞和类人胶原蛋白的浓度均显著高于其他两种控氧方式,充入富氧空气的细胞和类人胶原蛋白的浓度均最低;诱导强度对细胞生长和蛋白质表达的影响非常大,当细胞浓度达47±2g/L(DCW)时,开始升温至42℃进行诱导,最佳条件为42℃保温2～3h,然后降温至39℃继续培养5～6h。结论通过优化重组大肠杆菌分批补料培养生产类人胶原蛋白的工艺,使得细胞和类人胶原蛋白浓度分别达68.94g/L(DCW)和13.02g/L。Aim Process control of recombinant Escherichia coli in fed-batch culture should be optimized. Methods The effects of specific growth rate, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated to optimize fermentation conditions.Results The results showed that specific growth rate affects human-like collagen production greatly and its optimum value was (0.15)~0.20 h^(-1); cell density and concentration of human-like collagen controlled by enhancing the pressure of fermentation bioreactor were higher than that of the other two methods and those controlled by supplying oxygen-enriched air were lowest; the effects of different inducement strength on cell growth and protein expression were very great, when cell density reached to 47±2 g/L(DCW), induction should begin and the optimum inducement conditions were to cultivate at 42 ℃ for about 2~3 h and to continue cultivate at 39 ℃ for 5~6 h.Conclusion The concentration of biomass and human-like collagen could reach to 68.94 g/L (DCW) and 13.02 g/L respectively after optimizing the process control of recombinant Escherichia coli to produce human-like collagen.

关 键 词：类人胶原蛋白 重组大肠杆菌 半分批.补料培养 比生长速率 控氧方式 温度诱导 

分 类 号：Q939[生物学—微生物学]