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Improvement in recombinant protein production in ppGpp‐deficient Escherichia coli
Author(s) -
Dedhia Neilay,
Richins Richard,
Mesina Archie,
Chen Wilfred
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19970220)53:4<379::aid-bit4>3.0.co;2-k
Subject(s) - recombinant dna , escherichia coli , intracellular , protein biosynthesis , exponential growth , messenger rna , stringent response , biochemistry , biology , biosynthesis , enterobacteriaceae , cell growth , strain (injury) , fed batch culture , overproduction , chemistry , enzyme , gene , fermentation , mathematical analysis , mathematics , anatomy
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 fed‐batch 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 CAT‐specific 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. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 379–386, 1997.