Improved fermentation method for production of heterologous...

C - Chemistry – Metallurgy – 12 – P

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C12P 21/00 (2006.01) C12N 1/00 (2006.01) C12N 9/22 (2006.01) C12N 15/67 (2006.01) C12N 15/74 (2006.01)

Patent

CA 2426084

Fed-batch or continuous process for producing heterologous peptides, polypeptides or proteins in lactic acid bacteria, comprising the steps of (i) constructing a recombinant lactic acid bacterium comprising a nucleotide sequence coding for the heterologous peptide, polypeptide or protein, and appropriate regulatory nucleotide sequences such as regulatable promoters and signal peptides to control the expression of the coding sequence and the secretion of gene product, (ii) cultivating the recombinant bacterium under fed-batch or continuous cultivation conditions to express the gene, and (iii) harvesting the recombinant bacterium or the gene product. Preferably, the cultivation medium is a chemically defined or synthetic medium optionally supplemented with yeast extract.

Cette invention a trait un procédé, à écoulement discontinu ou continu, de production de peptides, de polypeptides ou de protéines hétérologues dans des bactéries d'acide lactique, lequel procédé consiste, (i), à produire une bactérie de recombinaison d'acide lactique renfermant une séquence nucléotidique codant le peptide, le polypeptide ou la protéine hétérologue, ainsi que des séquences nucléotidiques régulatrices appropriées telles que des promoteurs régulables et des peptides signal afin d'agir sur l'expression de la séquence codante et la sécrétion du produit génique, (ii), à cultiver la bactérie de recombinaison par culture à écoulement discontinu ou continu afin de produire le gène et, (iii), à recueillir la bactérie de recombinaison ou le produit génique. Le milieu de culture est, de préférence, un milieu chimiquement défini ou un milieu synthétique auquel est éventuellement adjoint un extrait de levure.

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