Method for immobilizing porcine pancreatic lipase
A technology of porcine pancreatic lipase and lipase, which is applied in the direction of immobilization on or in the inorganic carrier, can solve the problems of poor mechanical properties, low biocompatibility, and low molecular weight, and achieve stable structure, wide application prospects, and preparation The method is simple and efficient
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Embodiment 1
[0015] Add aminated silica particles (10 mg) into 0.1mol / L, pH=7.5 phosphate buffered saline (PBS) (10mL), ultrasonically disperse the particles in the solution, and pass nitrogen gas for 30min to remove the pipeline and Add 0.87 mL of 25% glutaraldehyde to the air in the flask under the protection of nitrogen, and then pass nitrogen for 5 minutes, stir magnetically at 25 °C for 2 hours, centrifuge to discard the supernatant, and wash the particles with 0.01 mol / L PBS solution. Remove residual glutaraldehyde. Add 2 mL of 4 mg / mL porcine pancreatic lipase (IPPL) solution to the carrier solution, place it in a constant temperature shaker at 40°C and shake it, and react for 8 h. After the reaction is completed, centrifuge to discard the supernatant, and add glycine ( 0.005 g) blocked for 30 min, washed by centrifugation, washed with PBS (0.01 mol / L, containing 0.001 mol / L EDTA (ethylenediaminetetraacetic acid) and 0.008 mol / L cysteine) and dried at a constant temperature of 40 °C...
Embodiment 2
[0017] Add aminated silica particles (10 mg) into 0.1mol / L, pH=7.5 phosphate buffered saline (PBS) (10mL), ultrasonically disperse the particles in the solution, and pass nitrogen gas for 30min to remove the pipeline and Add 0.87mL of 25% glutaraldehyde to the air in the flask under the protection of nitrogen, and then pass nitrogen for 5 minutes, stir magnetically at 25°C for 2 hours, centrifuge to discard the supernatant, and wash the particles with 0.01 mol / L PBS solution , to remove residual glutaraldehyde. In the carrier solution, add 1 mL of 4 mg / mL porcine pancreatic lipase (IPPL) solution, place it in a constant temperature shaker at 40 °C and shake it, and react for 8 h. After the reaction, centrifuge to discard the supernatant, and add glycine ( 0.005 g) blocked for 30 min, washed by centrifugation, washed with PBS (0.01 mol / L, containing 0.001 mol / L EDTA (ethylenediaminetetraacetic acid) and 0.008 mol / L cysteine) and dried at a constant temperature of 40 °C , Amina...
Embodiment 3
[0019] Add aminated silica particles (10 mg) into 0.1mol / L, pH=9 phosphate buffered saline (PBS) (10mL), ultrasonically disperse the particles in the solution evenly, pass nitrogen gas for 30min to remove the pipeline and Add 0.87mL of 25% glutaraldehyde to the air in the flask under the protection of nitrogen, and then pass nitrogen for 5 minutes, stir magnetically at 25°C for 2 hours, centrifuge to discard the supernatant, and wash the particles with 0.01 mol / L PBS solution , to remove residual glutaraldehyde. Add 2 mL of 4 mg / mL porcine pancreatic lipase (IPPL) solution to the carrier solution, place it in a constant temperature shaker at 40°C and shake it, and react for 8 h. After the reaction, centrifuge to discard the supernatant, and add glycine (0.005 g) Block for 30 min, wash by centrifugation, wash the particles with PBS (0.01 mol / L, containing 0.001 mol / L EDTA (ethylenediaminetetraacetic acid) and 0.008 mol / L cysteine) and dry at a constant temperature of 40°C. Ami...
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