CS218890B1 - A method for isolating neutral proteases - Google Patents

Abstract

The invention relates to the chromatographic isolation of neutral proteases containing zinc using new insoluble carriers. Proteases separated by these carriers are used in the field of medicine, in the food industry and in the beverage industry. Isolation is carried out by bringing weakly basic solutions containing neutral proteases into contact with heterocyclic carriers, the heterocycles containing at least one endocyclic tertiary nitrogen atom with a free electron pair. In order to separate other proteins, elution is carried out with buffer solutions of higher ionic strength and in a weakly alkaline environment, and neutral zinc-containing proteases are eluted in a weakly acidic environment. Dialysis is carried out using a C.1% calcium acetate solution

Description

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Claims (5)

- 12 - 218 890 OBJECT OF THE INVENTION A process for the isolation of zinc-containing neutral proteases, characterized in that the solutions of pS 7 to 10, preferably 8.5 to 9.5, containing neutral proteases, are contacted with carriers comprising heterocycles, either column or batch. in a manner wherein the heterocycles must contain at least one endocyclic, free electron pair tertiary nitrogen, the other proteins are eluted with buffer solutions, and the ionic strengths of pH 8.5 to 9.5 and the zinc-containing proteases bound to carriers containing heterocycles are eluted with pH buffer. 4 to 7 with the addition of inorganic salts, dialyzing with 0.01% calcium acetate solution and lyophilizing. 2. A process as claimed in claim 1, wherein a zinc-containing, neutral protease-containing bio-material is a material of bacterial origin, in particular a fermentation broth culture liquid or a dry product thereof. purified solutions. 3. A method according to claim 1 or 2, characterized in that the fermentation media of bacteria such as B. amylo-liquefaciens, B. cerusus, B. licheninformis, B. megaterium, B. subtilis, B.thermoproteolyticus, B.thuringiensis aB are used. . pumylis, which are obtained by microbial fermentation. 4. A process as claimed in claim 1, wherein the heterocyclic-containing carriers are copolymers of hydroxyalkyl methacrylates (C2-C8-alkyl) with enhancers selected from the group of divinyl or polyvinyl monomers of the methacrylate type. 5. The process of claim 1 wherein the heterocyclic-containing carriers are copolymers containing 5, 6 or more membered heterocycles, preferably imidazole benzimidazole and derivatives thereof, as well as pyridine derivatives. Recognized by the invention on the basis of the results of an expert review by the Office of Invention and Patenting, Berlin, DD.