Frozen gel embedding method for enzyme
A technology of cryogel and enzyme liquid, which is applied in the direction of fixing on/in the organic carrier, can solve the problems of leakage and reduce enzyme activity, etc., and achieve the effect of simple process, high catalytic activity and strong resistance to organic solvents
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Embodiment 1
[0025] First, weigh urease powder and dissolve it in boric acid buffer solution to make 0.2g / L enzyme solution for later use, add 5mL N-isopropylacrylamide with a concentration of 70g / L to 2mL enzyme solution, add 0.035g polyethylene glycol 200 diacrylate, 0.04mL hydrogen peroxide with a concentration of 30%, and then placed in a tetrafluoroethylene tray, placed in a low-temperature constant temperature bath at -20°C for 3 hours; placed in an ultraviolet light (wavelength 365nm) polymerization instrument for 2 minutes, The obtained gel was soaked in distilled water for 2 days, the water was changed every 4 hours, and after 3 times, the water was changed every 12 hours, and then vacuum-dried at 40°C to obtain the cryogel-embedded enzyme. The measured activity yield of the enzyme was 95%, the gel yield was 92%, and the gel swelling rate was 35%. After storage at 4°C for 28 days, the activity of the enzyme retained 94.8% of the original activity.
[0026] Wherein, the activity y...
Embodiment 2
[0028] First, weigh urease powder and dissolve it in boric acid buffer solution to make 1g / L enzyme solution for later use. Add 5mL acrylamide with a concentration of 70g / L to 2mL enzyme solution, add 0.07g polyethylene glycol 400 diacrylate, 0.06 Put hydrogen peroxide with a concentration of 30% in mL, then place it in a tetrafluoroethylene dish, place it in a low-temperature thermostat at -25°C for 2 hours; Soak for 3 days, change the water every 3 hours, and change the water every 12 hours after 4 times, and then vacuum-dry at 40°C to obtain the cryogel-embedded enzyme. The enzyme activity yield was measured to be 89%, the gel yield was 94%, and the gel swelling rate was 34%. After storage at 4°C for 28 days, the activity of the enzyme retained 80% of the original activity.
Embodiment 3
[0030] First, weigh papain powder and dissolve it in phosphate buffer solution to prepare 2.5g / L enzyme solution for later use, add 5mL acrylamide with a concentration of 100g / L to 2mL enzyme solution, add 0.10g polyethylene glycol 200 dimethyl Acrylic acid ester, 0.09mL of hydrogen peroxide with a concentration of 30%, and then placed in a tetrafluoroethylene tray, placed in a low-temperature thermostat at -15°C for 5h; placed in an ultraviolet light (wavelength 365nm) polymerizer for 4min to obtain The gel was soaked in distilled water for 2 days, the water was changed every 4 hours, and the water was changed every 12 hours after 3 times, and then vacuum-dried at 40°C to obtain the cryogel-embedded enzyme. The measured activity yield of the enzyme was 95%, the gel yield was 99%, and the gel swelling rate was 29%. After storage at 4°C for 28 days, the enzyme activity retained 95.7% of its original activity.
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