Jung G, Denèfle P, Becquart J, Mayaux J F
Rhône-Poulenc Santé, Centre de Recherches de Vitry, Vitry-sur-Seine, France.
Ann Inst Pasteur Microbiol. 1988 Jan-Feb;139(1):129-46. doi: 10.1016/0769-2609(88)90100-7.
A high-productivity process has been developed for the production of mature human interleukin-1 beta (IL-1 beta) from recombinant Escherichia coli strains. Conditions were found that allow high IL-1 beta expression levels in high cell density cultures. Improved fed-batch fermentation strategies are described which include maintenance of glucose and acetate concentrations below 1 g/l and sparging the fermentor with an O2-enriched air supply. Using the E. coli tryptophan promoter control of transcription, a 2.2 g/l production level of IL-1 beta was achieved in E. coli B at cell densities of 55 g dry weight per litre. Another genetic construction involving the bacteriophage lambda cIts-PR expression cassette allowed a similar IL-1 beta production level (1.9 g/l) in E. coli E103S, albeit at a lower cell density (30 g/l). A simplified procedure allowing the purification of fully active IL-1 beta is also presented.
已开发出一种用于从重组大肠杆菌菌株生产成熟人白细胞介素-1β（IL-1β）的高生产率工艺。发现了在高细胞密度培养中允许高IL-1β表达水平的条件。描述了改进的补料分批发酵策略，包括将葡萄糖和乙酸盐浓度维持在1 g/l以下，并用富氧空气供应对发酵罐进行鼓泡。利用大肠杆菌色氨酸启动子控制转录，在大肠杆菌B中，细胞密度为每升55 g干重时，IL-1β的产量达到2.2 g/l。另一种涉及噬菌体λ cIts-PR表达盒的基因构建在大肠杆菌E103S中实现了相似的IL-1β产量水平（1.9 g/l），尽管细胞密度较低（30 g/l）。还介绍了一种简化的方法，可用于纯化具有完全活性的IL-1β。
