Methods for preparing and using peroxidase electrode
A technology of peroxidase and capsicum peroxide, which is applied in the preparation and application of peroxidase electrodes, can solve the problems of short life and poor stability of enzyme electrodes, and achieve high biocompatibility and strong stability , the effect of strong film-forming ability
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Embodiment approach 1
[0024] Commercially available dried red peppers were crushed through 80 meshes, weighed 20.0g of chili powder, added 200mL of distilled water, ultrasonicated at low temperature (0°C) for 20min, and were centrifuged at 4000r / min for 15min to obtain the above Serum. Slowly add ammonium sulfate with a saturation of 0.4 to the supernatant extracted above at 0°C under constant stirring (add 226.0 g of ammonium sulfate per 1000 mL of filtrate), and put it in the refrigerator for 24 hours. Aspirate the supernatant, centrifuge the cloudy solution below at a centrifugal speed of 3000r / min for 15min, discard the precipitate and combine the supernatant. Add ammonium sulfate with a saturation of 0.8 at 0°C to the supernatant (add 258 g of ammonium sulfate per 1000 mL of supernatant) and stir until it is completely dissolved, and put it in the refrigerator for 24 hours. Aspirate the supernatant, centrifuge the precipitate at a centrifugal speed of 13000r / min for 20min, discard the superna...
Embodiment approach 2
[0026] The dry red pepper sold in the market was used as raw material and crushed to 85 mesh, weighed 25g of dried red pepper powder crushed to 85 mesh, added 250mL of distilled water, ultrasonicated at 0°C for 30min, suction filtered, centrifuged to obtain a crude extract, first The extracted crude extract was dialyzed with an ammonium sulfate solution with a saturation of 0.4, and the supernatant was obtained after centrifugation, and then dialyzed with an ammonium sulfate solution with a saturation of 0.8, and the precipitate after centrifugation was added to 1 times the volume of -15 ℃ acetone fractionation for dialysis, the solution after dialysis is filtered by an ultrafiltration membrane with a molecular weight cut-off of 5000 to 10000, and the capsicum peroxidase obtained at 4 ℃; the measured concentration of capsicum peroxidase is 8.56g / L, the activity is 15.53u / g. will be 1.0cm 2 Soak large and small platinum pieces in 1moL / L NaOH solution for 0.5h, then take out th...
Embodiment approach 3
[0028] The dried red peppers sold in the market were used as raw materials and crushed to 100 mesh, weighed 50 g of dried red pepper powder crushed to 100 mesh, added 500 mL of distilled water, ultrasonicated at 0°C for 40 minutes, suction filtered, and centrifuged to obtain a crude extract. The extracted crude extract was dialyzed with an ammonium sulfate solution with a saturation of 0.4, and the supernatant was obtained after centrifugation, and then dialyzed with an ammonium sulfate solution with a saturation of 0.8, and the precipitate after centrifugation was added to 2 times the volume of acetone fractionation at -16°C for dialysis, the solution after dialysis is filtered with an ultrafiltration membrane with a molecular weight cut-off of 10,000, and the capsicum peroxidase obtained at 6°C has a measured concentration of 10.25g / L and an activity of 23.3 u / g. 1.5cm 2 Soak large and small platinum pieces in 1moL / L NaOH solution for 1 hour, then take out the cotton ball d...
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