High-cell density fermentation studies of recombinant Escherichia coli strains expressing human interleukin-1 beta.
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.
Jung
G
G
Rhône-Poulenc Santé, Centre de Recherches de Vitry, Vitry-sur-Seine, France.
Denèfle
P
P
Becquart
J
J
Mayaux
J F
JF
D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D007375:Interleukin-1; D011994:Recombinant Proteins