Use of high density cultures ofEscherichia colifor high level production of recombinantPseudomonas aeruginosaexotoxin A

✍ Scribed by R. Fass; M. Walle; A. Shiloach; A. Joslyn; J. Kaufman; J. Shiloach

• Publisher: Springer
• Year: 1991
• Tongue: English
• Weight: 597 KB
• Volume: 36
• Category: Article
• ISSN: 1432-0614
• DOI: 10.1007/bf00164700

✦ Synopsis

An efficient fermentation method for the production of two modified recombinant Pseudomonas aeruginosa exotoxin As cloned in Escherichia coli BL21(ADE3) was developed. Cell densities of 16-30 g dry weight/1 were found to be most suitable for the induction of protein synthesis, which was under the isopropyl fl-D-thiogalactopyranoside (IPTG)-inducible T7 expression system. A concentration of 0.6 mM I P T G and induction time of 90 min were found to give the best results for production of the modified toxins. Using this procedure, gram amounts of the proteins were obtained in a 3-1 bench-top fermentor. The high density growth of the bacteria did not impair the integrity of the proteins and did not interfere with the purification procedure.