Chymosin recovery and purification

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 9/64 (2006.01)

Patent

CA 2058453

2058453 9015865 PCTABS00003 This invention provides a method of recovering and purifying chymosin from fermentation used for microbial production of chymosin. The fermentation mixture is filtered to provide a filtrate containing the chymosin. The filtrate is first prepared by either adjusting it to a low pH, such as about 2 and/or adjusting the salt concentration of the filtrate to at least about 2M. The aqueous filtrate mixture is contacted with a phenyl-sepharose resin, which selectively binds the chymosin from the filtrate. The filtrate remaining after the contact with the phenyl-sepharose resin is discarded, and the phenyl-sepharose resin column is eluted with water or dilute salt solution to obtain substantially pure chymosin in a single elution step.

L'invention concerne un procédé pour récupérer et purifier de la chymosine lors d'une fermentation pour la production microbienne de chymosine. Le mélange de fermentation est filtré de manière à réaliser un filtrat contenant ladite chymosine. On prépare d'abord ledit filtrat en l'ajustant à un pH faible, environ 2, et/ou en ajustant la concentration de sel dans ledit filtrat à au moins environ 2M. Le mélange aqueux du filtrat est mis en contact avec une résine phényle-sépharose, qui lie sélectivement ladite chymosine provenant du filtrat. Le filtrat qui reste après le contact avec ladite résine phényle-sépharose est rejeté, la colonne de résine phényle-sépharose étant éluée avec de l'eau ou avec une solution de sel dilué de manière à obtenir de la chymosine essentiellement pure lors d'une seule étape d'élution.

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