Method for preparing mixed immobilized glucose oxidase/catalase microspheres
A technology of glucose oxidase and catalase, which is applied in the direction of immobilized on/in organic carriers, multi-enzyme systems, etc., can solve problems such as the difficulty in realizing the full utilization of amino groups, and achieve sustainable development and eliminate damage , The effect of improving industrial production efficiency
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Embodiment 1
[0024] A method for preparing co-immobilized glucose oxidase / catalase microspheres, the specific method steps are as follows:
[0025] (1) Preparation of chitosan-arginine anionic microspheres
[0026] According to the article "Chitosan-Arginine Resin Immobilization of Chymotrypsin and Its Properties" published in "Applied Chemistry" Volume 26 Issue 7 in July 2009, chitosan-arginine anionic microspheres were prepared .
[0027] (2) Prepare GOD and CAT mixed solution
[0028] After the first step (1) is completed, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 3~8 according to the ratio of GOD:CAT activity ratio of 1:1~6, The lysates containing GOD and CAT were collected. For the collected mixed solution containing GOD and CAT, it was used to prepare co-immobilized GOD / CAT.
[0029] (3) Co-immobilization of GOD and CAT
[0030] After the (2) step was completed, the mass (g) of the chitosan-arginine anion microspheres...
Embodiment 2
[0034] A method for preparing co-immobilized glucose oxidase / catalase microspheres, same as in Example 1, wherein:
[0035] In the (2) step, the activity ratio of GOD:CAT is 1: 4, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 5;
[0036] In the (3) step, the quality of the chitosan-arginine anion microsphere prepared in the (1) step: the volume ratio of the mixed solution containing GOD and CAT collected in the (2) step is 1g: 5mL, After 45 minutes of stirring and adsorption, the glutaraldehyde volume concentration is 2.5% glutaraldehyde solution: the volume ratio of the mixed solution containing GOD and CAT is 1: 7, and the reaction is 2.5 hours at 7 ° C;
[0037] In step (4), wash 4 times with 0.2 mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 5, freeze-dry for 27 hours at a temperature of -45° C. and a vacuum of 35 Pa.
Embodiment 3
[0039] A method for preparing co-immobilized glucose oxidase / catalase microspheres, same as in Example 1, wherein:
[0040] In the (2) step, the activity ratio of GOD:CAT is 1: 6, GOD and CAT are dissolved in the 0.2mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 8;
[0041] In the (3) step, the quality of the chitosan-arginine anion microsphere prepared in the (1) step: the volume ratio of the mixed solution containing GOD and CAT collected in the (2) step is 1g: 8mL, After 60 minutes of stirring and adsorption, the glutaraldehyde volume concentration was 2.5% glutaraldehyde solution: the volume ratio of the mixed solution containing GOD and CAT was 1: 10, and the reaction was carried out at 10° C. for 3 hours;
[0042] In step (4), wash 5 times with 0.2 mol / L disodium hydrogen phosphate-citric acid buffer solution of pH 8, freeze-dry for 30 hours at a temperature of -50° C. and a vacuum of 50 Pa in a freeze dryer.
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