Preparation of water-insoluble enzyme compositions
AssigneeKali-Chemie Aktiengesellschaft
InventorsGuenter Weidenbach, Dirk Bonse
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Abstract
Enzymes are covalently bonded to porous inorganic supports by a process wherein maximum activity of enzyme in the water-insoluble enzyme composition is achieved with the lowest possible amount of enzyme. The process involves selecting an inorganic carrier having a most frequent pore-diameter that results in highest enzymatic activity possible per weight unit of the insoluble enzyme composition at the given enzyme concentration within the insoluble enzyme composition, and contacting the selected inorganic carrier with an enzyme solution containing an amount of enzyme which is sufficiently low so that the enzyme in the insoluble enzyme composition has a specific activity which is substantially the same as that of the enzyme in solution in the free state.
Claims27
A process for preparing a water-insoluble enzyme composition wherein an enzyme is covalently bonded to an inorganic support material whereby a maximum activity of the insoluble enzyme composition is achieved with the lowest possible amount of enzyme, said process comprising the steps of:
- (a) selecting an inorganic support material having the most frequent pore-diameter which produces an enzyme composition having the highest absolute activity when a plurality of inorganic support materials having different most frequent pore-diameters are each individually contacted with an enzyme solution containing a given concentration of enzyme, said most frequent pore-diameter producing the highest absolute activity being independent of the enzyme concentration in the solution;
- (b) selecting an aqueous enzyme solution containing the concentration of enzyme which produces a composition that has a relative activity of substantially 100% when the support material of step (a) is contacted with a plurality of enzyme solutions having different concentrations of enzyme;
- (c) contacting the support material of step (a) with the enzyme solution of step (b) whereby a portion of the enzyme from the solution is taken up by the support material to produce a water-insoluble enzyme composition;
- and
- (d) separating the resulting water-insoluble enzyme composition from the remaining solution.
A process of producing a water-insoluble enzyme composition wherein an enzyme is covalently bonded to an inorganic support material whereby maximum activity of the insoluble enzyme composition is achieved with the lowest possible amount of enzyme comprising the steps of:
- (a') contacting at least 2 different inorganic support materials capable of covalently bonding the enzyme, each having a different most frequent pore-diameter, each with at least two different solutions of the enzyme, each having a different enzyme content to form a water-insoluble enzyme composition;
- (b') determining the enzymatic activity per weight unit of the enzyme compositions obtained in step (a');
- (c') comparing enzymatic activities of the enzyme compositions determined in step (b') and determining the inorganic support material having the most frequent pore-diameter that yields the enzyme composition having the highest enzymatic activity for the at least two different solutions of enzyme;
- (d') contacting the inorganic support material having the most frequent pore-diameter determined in step (c') with a plurality of solutions containing different amounts of the enzyme to form a plurality of water-insoluble enzyme compositions;
- (e') determining the specific activity of each of the plurality of enzyme compositions obtained in step (d');
- (f') comparing the specific activity of each water-insoluble enzyme composition determined in step (e') with the specific activity of the enzyme in the free state contained in each of said plurality of enzyme solutions to determine the relative activity of each water-insoluble enzyme composition;
- (g') determining from the different amounts of enzyme in the enzyme solutions the amount which yields a water-insoluble enzyme composition having a relative activity of substantially 100%;
- (h') contacting the inorganic support material having the most frequent pore-diameter as determined in step (c') with a solution containing the amount of enzyme as determined in step (g') to form a water-insoluble enzyme composition;
- and
- (i') separating the water-insoluble enzyme composition from the remaining solution.
Dependencies are inferred from the claim text.