A kind of horseradish peroxidase immobilization method
A technology of horseradish peroxidase and methacryloxypropyltrimethoxysilane, which is applied in the direction of immobilized enzymes, oxidoreductases, biochemical equipment and methods, etc., can solve the problem of difficult recycling and reuse rate Low-level problems, to achieve the effect of mild conditions, high immobilization efficiency, and improved stability
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Embodiment 1
[0030] 1. Measure 200 μL of methacryloxypropyltrimethoxysilane (MEMO) into 10 ml of 95% ethanol, and dropwise add 10% citric acid water to adjust the pH to 2-3. Weigh 100 mg of micron-scale expanded perlite, add it to the above reaction system, and soak it for 0.5-1 h. The above reaction mixed liquid is filtered, washed and dried to obtain the modified expanded perlite powder.
[0031]2. Weigh 10 mg of horseradish peroxidase, suspend it in a round-bottomed flask containing 5 mL of phosphate buffer solution (pH=7.1), and stir for about 30 min, and then dropwise add 2 mL of methacrylic anhydride to the round-bottomed flask at a rate of about is 1ml / min. The above suspension is centrifuged, precipitated and freeze-dried to obtain methacrylylated horseradish peroxidase.
[0032] 3. 100 mg of the obtained modified expanded perlite and 10 mg of methacrylated horseradish peroxidase were mixed and dispersed in 5 ml of acrylic acid aqueous solution with a mass fraction of 2%, and a p...
Embodiment 2
[0034] 1. Measure 400 μL of methacryloxypropyltrimethoxysilane (MEMO) into 10 ml of 95% ethanol, and add 10% citric acid water dropwise to adjust the pH to 2-3. 200 mg of micron-scale expanded perlite was weighed and added to the above reaction system, and soaked for 0.5-1 h. The above reaction mixed liquid is filtered, washed and dried to obtain the modified expanded perlite powder.
[0035] 2. Weigh 10 mg of horseradish peroxidase, suspend it in a round-bottomed flask containing 5 mL of phosphate buffer solution (pH=7.1), and stir for about 30 min, and then add 2 mL of base acrylic anhydride dropwise to the round-bottomed flask at a rate of about 1ml / min. The above suspension is centrifuged, precipitated and freeze-dried to obtain methacrylylated horseradish peroxidase.
[0036] 3. 200 mg of the obtained modified expanded perlite and 10 mg of methacrylated horseradish peroxidase were mixed and dispersed in an acrylic acid aqueous solution with a mass fraction of 2%, and a ...
Embodiment 3
[0038] 1. Measure 500 μL of methacryloxypropyltrimethoxysilane (MEMO) into 10 ml of 95% ethanol, and dropwise add 10% citric acid water to adjust the pH to 2-3. 250 mg of micron-scale expanded perlite was weighed and added to the above reaction system, and soaked for 0.5-1 h. The above reaction mixed liquid is filtered, washed and dried to obtain the modified expanded perlite powder.
[0039] 2. Weigh 10 mg of horseradish peroxidase, suspend it in a round-bottomed flask containing 5 mL of phosphate buffer solution (pH=7.1), and stir for about 30 min, and then dropwise add 2 mL of methacrylic anhydride to the round-bottomed flask at a rate of about is 1ml / min. The above suspension is centrifuged, precipitated and freeze-dried to obtain methacrylylated horseradish peroxidase.
[0040] 3. 250 mg of the obtained modified expanded perlite and 10 mg of methacrylated horseradish peroxidase were mixed and dispersed in an aqueous solution of acrylic acid with a mass fraction of 2%, a...
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