Direct expression of peptides into culture media

Details Direct expression of peptides into culture media Direct expression of peptides into culture media

C - Chemistry, Metallurgy

12

N

C12N 1/00 (2006.01) C07K 14/585 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/63 (2006.01) C12N 15/70 (2006.01) C12P 21/02 (2006.01) C12P 21/06 (2006.01)

Patent

CA 2285658

Expression systems are disclosed for the direct expression of peptide products into the culture media where genetically engineered host cells are grown. High yield was achieved with novel vectors, a special selection of hosts, and/or fermentation processes which include careful control of cell growth rate, and use of an inducer during growth phase. Special vectors are provided which include control regions having multiple promoters linked operably with coding regions encoding a signal peptide upstream from a coding region encoding the peptide of interest. Multiple transcription cassettes are also used to increase yield. The production of amidated peptides using the expression systems is also disclosed.

L'invention concerne des systèmes d'expression permettant l'expression directe de produits peptidiques dans un milieu de culture, dans lequel on fait croître des cellules hôtes génétiquement manipulées. On a ainsi pu obtenir un rendement élevé grâce à de nouveaux vecteurs, à une sélection spéciale des hôtes, à l'utilisation d'un inducteur au cours de la phase de croissance, et/ou à des processus de fermentation consistant à surveiller de près la vitesse de croissance des cellules. L'invention concerne également des vecteurs particuliers, comprenant les régions de régulation qui présentent plusieurs promoteurs liés de manière opérationnelle aux régions codantes qui codent un peptide signal en amont, à partir d'une région codante qui code le peptide d'intérêt. On utilise également plusieurs cassettes de transcription pour améliorer le rendement. L'invention concerne enfin la production de peptides amidifiés au moyen desdits systèmes d'expression.

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