Preparation method of multi-level structure immobilized enzyme
A technology for immobilizing enzymes and preparation steps, applied in the field of bioengineering, can solve the problems of high enzyme activity and stable immobilization, and achieve good activity and operational stability, high enzyme load, and prevent the loss of enzymes. Effect
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Embodiment 1
[0018] (1) Preparation of alumina-silicon oxide composite film
[0019] Dissolve 1 g of polyvinyl ether-polypropylene ether-polyvinyl ether block copolymer (P123) in a mixture of 5 g of ethanol and 1.2 g of 0.2 mol / L hydrochloric acid at room temperature, add 0.03 g of toluene, and stir for 2 hours. 2.08 g of ethyl orthosilicate was added dropwise into the solution, and stirred for 1 hour to obtain a silicon oxide precursor solution. An anodic aluminum oxide film with a diameter of 47mm and a pore size of 200nm was immersed in the silicon oxide precursor solution, and volatilized under vacuum for 1 hour. The aluminum oxide film was taken out and naturally crystallized for 1 day. 180mL ethanol was refluxed three times for 24 hours to extract the block copolymer P123. Dry at room temperature.
[0020] (2) Filtration and adsorption of hemoglobin
[0021] Take 5 mL of 0.1 mol / L disodium hydrogen phosphate and 90 mL of 0.1 mol / L potassium dihydrogen phosphate solution to prepar...
Embodiment 2
[0030] (1) Preparation of alumina-silicon oxide composite film
[0031] Dissolve 1 g of polyvinyl ether-polypropylene ether-polyvinyl ether block copolymer (P123) in a mixture of 5 g of ethanol and 1 g of 0.2 mol / L hydrochloric acid at room temperature, add 0.15 g of toluene, and stir for 2 hours. 2.08 g of ethyl orthosilicate was added dropwise into the solution, and stirred for 1 hour to obtain a silicon oxide precursor solution. An anodic aluminum oxide film with a diameter of 47mm and a pore size of 200nm was immersed in the silicon oxide precursor solution, and volatilized under vacuum for 1 hour. The aluminum oxide film was taken out and naturally crystallized for 1 day. 180mL ethanol was refluxed three times for 24 hours to extract the block copolymer P123. Dry at room temperature.
[0032] (2) Filtration and adsorption of myoglobin
[0033]Take 10 mL of 0.1 mol / L disodium hydrogen phosphate and 90 mL of 0.1 mol / L potassium dihydrogen phosphate solution to prepare a...
Embodiment 3
[0042] (1) Preparation of alumina-silicon oxide composite film
[0043] Dissolve 0.687g of polyvinyl ether-polypropylene ether-polyvinyl ether block copolymer (P123) in a mixture of 4.8g of ethanol and 1.2g of 0.2mol / L hydrochloric acid at room temperature, add 0.01g of toluene, and stir for 2 hours . 2.08 g of ethyl orthosilicate was added dropwise into the solution, and stirred for 1 hour to obtain a silicon oxide precursor solution. An anodic aluminum oxide film with a diameter of 47mm and a pore size of 200nm was immersed in the silicon oxide precursor solution, and volatilized under vacuum for 1 hour. The aluminum oxide film was taken out and naturally crystallized for 1 day. 180mL ethanol was refluxed three times for 24 hours to extract the block copolymer P123. Dry at room temperature.
[0044] (2) Filtration and adsorption of trypsin
[0045] Take 100mL of 0.1mol / L Tris and 11.4mL of 0.1mol / L hydrochloric acid solution to prepare a pH 9 buffer solution to prepare ...
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