A kind of preparation method of magnetically immobilized lipase that can be used in organic medium
A technology for immobilizing lipase and lipase, applied in the directions of biochemical equipment and methods, immobilization on or in inorganic carriers, enzymes, etc.
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Embodiment 1
[0050] In the preparation method of magnetically immobilized lipase provided by the invention, the magnetic Fe 3 o 4 / P(St-AA) nanospheres were synthesized by the following method:
[0051] Step 1. Weigh 1059.5 mg of iron acetylacetonate and add it to 20 mL of diphenyl ether, then add 2 mL of oleylamine, 1 mL of ethylene glycol, and 2 mL of oleic acid, stir evenly, and react for 0.5 h at 200 ° C in a nitrogen atmosphere , and then reflux at 250° C. for 2 hours, stop heating, cool to room temperature, magnetically separate the black product, and wash with ethanol three times to obtain ferric oxide nanoparticles coated with oleic acid-oleylamine;
[0052] Step 2. Add 36mL sodium lauryl sulfate solution (mass concentration, 0.1mol / L) in the iron ferric oxide nano-particle of oleic acid-oleylamine wrapping obtained in step 1, after stirring and mixing, then Disperse under ultrasonic conditions for 15 minutes to prepare a magnetic ferric oxide emulsion;
[0053] Step 3. Add 90 m...
Embodiment 2
[0067] Magnetic Fe 3 o 4 The preparation of / P(St-AA) nanospheres is the same as in Example 1.
[0068] Magnetic Fe 3 o 4 The operation process of / P(St-AA) nanosphere immobilized lipase is as follows:
[0069] Step 1. The freeze-dried lipase powder derived from porcine pancreas is prepared into a 1 mg / mL lipase solution with pH=7.4 phosphate buffer, and stored in a refrigerator at 4°C for later use;
[0070] Step 2. take by weighing the magnetic Fe that 100mg above-mentioned method makes 3 o 4 / P(St-AA) nanospheres, sequentially added 191.7mg 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 57.6mg N-hydroxysuccinimide and 20mL morpholine Ethansulfonic acid buffer solution (0.1mol / L, pH=6.0), ultrasonically dispersed for 2min, and reacted at 25°C for 3h, the obtained product was magnetically separated and washed to obtain activated magnetic polystyrene nanospheres.
[0071] Step 3. Add 40mL of the above-mentioned lipase solution to the above-mentioned activat...
Embodiment 3
[0075] In the preparation method of magnetically immobilized lipase provided by the invention, the magnetic Fe 3 o 4 / PS-CH 2 Cl nanospheres were synthesized by the following method:
[0076] According to step 1 and step 2 of Example 1, a magnetic iron ferric oxide emulsion was prepared; 72 mg of sodium lauryl sulfate and 5 mL of p-chloromethyl styrene were added successively to 24 mL of water, and after stirring and mixing evenly, the Disperse for 20 minutes to prepare a p-chloromethylstyrene monomer emulsion; mix the obtained magnetic iron ferric oxide emulsion and p-chloromethylstyrene monomer emulsion evenly, and add 30mg Potassium persulfate initiator, after reacting at 80°C for 2 hours, then add 30mg of potassium persulfate initiator, and react at 80°C for another 20h, after the reaction is completed, naturally cool to room temperature, and the obtained precipitate is subjected to magnetic separation, Magnetic Fe after washing and drying 3 o 4 / PS-CH 2 Cl nanospher...
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