SU443902A1 - The method of obtaining enzymes immobilized in the structure of polymerization gels - Google Patents

Description

one

FIELD OF THE INVENTION The invention relates to the biological, food, and food industry.

A method of producing enzymes immobilized in the structure of polysterization gels is known by mixing the enzyme solution with a mixture of acrylamide and l / N methylene bisacrylaide in the presence of radical polymerization catalysts.

C, however, with this method, a weak binding of small molecular weight enzymes acting on high molecular weight substrates to the gel is observed,

The proposed method makes it possible to improve the binding of enzymes with a mole, with a molecular weight of up to 50,000 s polyacrylamide gel.

This enzyme is pre-treated with solutions of anhydrous unsaturated acids. The latter, it is advisable to introduce in such

The concentrations that the ratio of KOjm4ecTBa alcohol workers of the enzyme and acid groups of the acid was 1: (). From anhydrite of unsaturated acids after, blows to use angi, dri, ditones of itaconic and maleic acids, and it is expedient to carry out the treatment of enzymes during 18-20 hours.

Angdrdda unsaturated acids at 4 ° C for 18-20 hours saturate the enzyme molecule with the cherry groups 1 and 1 without a significant decrease in the enzymatic activity.

Dy.difitsirovanny ferdyunts are subjected to copolymers with acrylic monomerales in the presence of catalysts radical by schmerization. The addition of solutions of unsaturated acids with gchhydride to enzymes: improves the properties of enzymes immobilized in the structure of polymerase-linked gels and improves “the structure of Iel, makes it more homogeneous, which 3 confirm the optical characteristics of the resulting gel of the resulting gel, Example. In order to saturate the enzyme with vinyl groups, the reaction mixture consists of equal volumes of trypsin solution with an activity of 8 U / ml (determined by Anson's method) and an itdonic acid anhydride solution with a concentration of 10 mgll., Which corresponds to the ratio of amino and anhydride groups I- (iiO -i40A, and heat the mixture at-4 ° C overnight. The activity of the enzymes is almost not reduced by the action of anhardric acid. To immobilize the enzymes, a polymerization mixture of the following composition is made: 7.6 h, reaction mixture A, 1.4 h , acrylamide solution and A /, A / - methylene bisacrylamide (90:10 ratio) and 0.5 h, solution / V, / V. Wyv tetramethydialene "The mixture is flushed with nitrogen for 15-30 minutes to remove oxygen, which inhibits the polserization, Then 0.5 h of ammonium persulphate solution is added to the mixture and praying is continued for 1-2 min. For a period of 10-15 h a fairly transparent gel block with enzymatic activity is formed. The gel block is crushed, repeatedly passed with a buffer solution and determine the activity of the enzyme in flowing waters, In gel 70-75 / 0 of the starting enzyme total is quenched. The activity of the enzyme enzyme is 2.5-3 E / g of gel, Pr, 1per 2. The conditions of example I are observed, but a solution of chymotrypsin with an activity of APE equal to 2060 is entered into the reaction mixture A. The activity of w / 1 mobile pharmac is 323 V / g gel, 443. . ... 902 4 Example 3, The conditions of example I are met, but the reaction mixture A is composed of a chymotrypsin solution with an ATPE activity equal to 4BOO 5 B / ml and a solution of al: i; drdd maleic acid with a ratio of amino and anhydride groups 1: 40, The immobilized enzyme activity is 140 U / g gel. PREPARING FROM ITS 2 ETENSH I. A method for producing enzymes immobilized in a polymerisation gel structure by solution of an enzyme with a mixture of acrylamide and l, i-methylenebisacrylamide In the presence of radical polymerization catalysts, characterized in that, in order to better bind enzyme to enzyme, polymer polymerization catalysts are different in that, in order to better bind enzyme to mole to enzyme polymerization catalysts, different from that, in order to better bind enzyme to molten enzyme, polymer polymerization catalysts are better in order to better bind enzyme polymerization catalysts. m weighing up to 50,000. The enzymes are pretreated with solutions of anhydrous unsaturated acids, .G 2, The method according to claim 1, characterized in that such concentrations of Angi solutions are used; the doses of the unsaturated acids are equal to the ratio of the number of amino groups of the enzyme and the Angi; the acid groups of the acid groups. I: (20-40) "35, 3, Method Jo PP, 1-2, characterized in that Ghz angi.rdrd of unsaturated Zhslot use Angdrdrd itaconic acid, 4, The method according to PPL-3, distinguishes 40 by the fact that angi; drvdov unsaturated acids use angi, dri; d maleic acid, 5, The method according to claims 1-4. This is due to the fact that the treatment of the i45 enzyme with anhydrous solutions of unsaturated acids is carried out in leaks. 18-20 hours at 4 ° C