Method for immobilizing enzyme based on magnetic nanoparticles modified by carboxymethyl starch
A technology of magnetic nanoparticles and carboxymethyl starch, applied in biochemical equipment and methods, fixed on or in inorganic carriers, fixed on/in organic carriers, etc., can solve the problem of increasing costs, difficult to recycle and repeat Utilization, difficult enzyme integrity and other problems, to achieve good thermal stability, improve coagulation phenomenon, high hydrophilic effect
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Embodiment approach 1
[0037] The preparation of sodium carboxymethyl starch:
[0038] Measure 30 ml of 80% ethanol solution in a three-necked flask, take 1.0 g of high amylose starch and fully dissolve it in the three-necked flask, add 2 g of sodium chloroacetate, ultrasonicate for 3 min, and after it is completely dissolved, add dropwise mol / L sodium hydroxide solution, kept stirring at a high speed, stopped dripping when the pH reached 10, continued to stir in an oil bath at 45 °C for 4 hours, cooled to room temperature after the reaction was completed, and washed and precipitated for many times Freeze-drying to obtain carboxymethyl starch particles.
[0039] Magnetic Fe 3 o 4 Preparation of nanoparticles:
[0040] Take 50 mL of deionized water in a three-necked flask, pass nitrogen gas with a purity >99.9% for 20 min, and add Fe at a molar ratio of 1:2 2+ and Fe 3+ , stirring continuously in an oil bath at a temperature of 50 °C until dissolved. After completely dissolving, add dropwise 1....
Embodiment approach 2
[0046] The present embodiment is roughly the same as embodiment 1, and the only difference is: in the preparation process of sodium carboxymethyl starch, the mass ratio of high amylose and sodium chloroacetate is 1:1.5, and the preparation temperature is 50°C.
[0047] In magnetic Fe 3 o 4 During the preparation of nanoparticles, Fe 2+ with Fe 3+ The mol ratio is 1:1.5, and the preparation temperature is 60°C.
[0048] In magnetic sodium carboxymethyl starch nanoparticles Fe 3 o 4 During the preparation of @CMS, sodium carboxymethyl starch and Fe 3 o 4 The mass ratio was 1:3, the preparation temperature was 90 °C, and the immobilization pH was 7.
[0049] Immobilization of enzyme Fe on magnetic sodium carboxymethyl starch nanoparticles 3 o 4 @CMS@BR (or Fe 3 o 4 @CMS @TLL or Fe 3 o 4 During the preparation process of @CMS@KDN), the glutaraldehyde concentration was 3.0% v / v, the addition amount was 2.0 ml, the crosslinking time was 2 h, the immobilization temperatu...
Embodiment approach 3
[0052] The present embodiment is roughly the same as Embodiment 1, and the only difference is: in the preparation process of sodium carboxymethyl starch, the mass ratio of high amylose and sodium chloroacetate is 1:1, and the preparation temperature is 40 °C.
[0053] In magnetic Fe 3 o 4 During the preparation of nanoparticles, Fe 2+ with Fe 3+ The molar ratio is 1:1, and the preparation temperature is 70 °C.
[0054] In magnetic sodium carboxymethyl starch nanoparticles Fe 3 o 4 During the preparation of @CMS, sodium carboxymethyl starch and Fe 3 o 4 The mass ratio was 1:4, the preparation temperature was 50 °C, and the pH of the immobilization environment was 10.
[0055] Immobilization of enzyme Fe on magnetic sodium carboxymethyl starch nanoparticles 3 o 4 @CMS@BR (or Fe 3 o 4 @CMS @TLL or Fe 3 o 4 During the preparation process of @CMS@KDN), the glutaraldehyde concentration was 2.5% v / v, the addition amount was 2.5 ml, the crosslinking time was 1 h, the immo...
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