Genetic manipulation of stationary-phase genes to enhance recombinant protein production in Escherichia coli

✍ Scribed by Chih-Hsiung Chou; George N. Bennett; Ka-Yiu San

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 749 KB
• Volume: 50
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19960620)50:6<636::aid-bit4>3.0.co;2-l

✦ Synopsis

Genetic manipulation of the host strain, by which cell physiology could be modulated, was exploited t o enhance recombinant protein production in Escherichia coli. The effects of an inactivated stationary-phase gene (rmf or katF) on recombinant protein production in strains with two different expression systems (the pH-inducible and the lac promoters) were investigated. An improvement of recombinant protein production in the katF mutant at low growth rates was observed for both expression systems. A fourfold and a 30% increase in the volumetric recombinant protein activity were observed for the pHinducible and the lac promoter system, respectively. The effect of the rmf mutation, on the other hand, depends on the expression system. A twofold increase in the volumetric recombinant protein activity was found for the pHinducible promoter system, but there was no improvementforthe lacpromoter system. Improvement in culture performance for slow-growing cultures may have an impact on the design strategy of the hosthector system used in fed-batch cultures, where the specific growth rate is usually slow. The information may also be useful for developing optimal host/vector gene expression systems for recombinant protein production. 0 1996