Improvement in recombinant protein production in ppGpp-deficient Escherichia coli

✍ Scribed by Neilay Dedhia; Richard Richins; Archie Mesina; Wilfred Chen

• Publisher: John Wiley and Sons
• Year: 1997
• Tongue: English
• Weight: 211 KB
• Volume: 53
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19970220)53:4<379::aid-bit4>3.0.co;2-k

✦ Synopsis

Absstract: Maintaining a metabolically productive state for recombinant Escherichia coli remains a central problem for a wide variety of growth-dependent biosynthesis. This problem becomes particularly acute under conditions of minimal cell growth such as fed-batch fermentations. In this, we investigated the possibility of manipulating the protein synthesis machinery of E. coli whereby synthesis of foreign proteins might be decoupled from cell growth. In particular, the effects of eliminating intracellular ppGpp on the synthesis of foreign proteins were studied in both batch and fed-batch operations. A significant increase in CAT production was observed from the ppGpp-deficient strain during both exponential and fedbatch phases. The increase in CAT production during exponential growth was accompanied by a simultaneous increase in CAT mRNA levels. Interestingly, CAT production was increased five-fold, while the level of CATspecific mRNA increased only three-fold. Thus, eliminating intracellular ppGpp appears to have increase the production of recombinant protein by increasing not only the pool sizes of CAT mRNA but also possible alternations in the post-transcriptional processes.