Efficient preparation method of low-value fish immunomdodulating peptides

[0024] Mix and homogenize golden thread fish myosin and 0.3Mmol NaCl (pH=6.8) at a concentration ratio of 1:10, put the homogenized myosin solution into a high-pressure micro-jet nano-homogenizer, and put the high-pressure nozzle The pore diameter was set to 5mm, the control treatment temperature was 35°C, the pressure of high-pressure homogenization treatment was 100MPa, the treatment time was 10s, and the number of cycles was 5 times. The mass ratio of protease to protein homogenate is 1:20, add trypsin (Wuxi Genencor Bioengineering Co., Ltd., 250u / mg), enzymolysis pH=7.5, enzymolysis temperature 40°C, stop enzymolysis after 60min, enzymolysis solution 90 ℃ water bath for 15 minutes to kill the enzyme; the enzymatic solution was centrifuged to remove the precipitate and then placed in the pulse electric field processor, connected to the ultrafiltration membrane, set the electromagnetic wave wavelength to 0.5cm, the electric field frequency to 4000MHz, and the radiation power t

[0026] Mix and homogenize the myosin from golden thread fish with 0.3Mmol NaCl (pH=6.8) at a concentration ratio of 1:10, put the homogenized myosin solution into a high-pressure micro-jet nano-homogenizer, and put the high-pressure The diameter of the nozzle was set to 5mm, the control treatment temperature was 35°C, the pressure of high-pressure homogenization treatment was 150MPa, the treatment time was 20s, and the number of cycles was 5 times. Under these conditions, the high-pressure homogenization treatment was carried out on the myosin solution of golden thread fish. Then add trypsin (Wuxi Genencor Bioengineering Co., Ltd., 250u / mg) at a mass ratio of trypsin to protein homogenate of 1:30, enzymatic hydrolysis pH=7.5, enzymatic hydrolysis temperature 40°C, stop enzymatic hydrolysis after 90min, enzymatic Inactivate the enzyme in a water bath at 90°C for 15 minutes. The enzymolysis solution was centrifuged to remove the precipitate and then placed in the pulse electric f

[0028]Mix and homogenize the myosin from golden thread fish with 0.3Mmol NaCl (pH=6.8) at a concentration ratio of 1:10, put the homogenized myosin solution into a high-pressure micro-jet nano-homogenizer, and put the high-pressure The aperture of the nozzle was set at 5mm, the treatment temperature was controlled at 35°C, the pressure of high-pressure homogenization treatment was 200MPa, the treatment time was 30s, and the number of cycles was 5 times. Under these conditions, the high-pressure homogenization treatment was performed on the myosin solution of golden thread fish. Then add trypsin (Wuxi Genencor Bioengineering Co., Ltd., 250u / mg) at a mass ratio of trypsin to protein homogenate of 1:40, enzymatic hydrolysis pH=7.5, enzymatic hydrolysis temperature 40°C, stop enzymatic hydrolysis after 1200min, enzymatic Inactivate the enzyme in a water bath at 90°C for 15 minutes. The enzymolyzed solution was centrifuged to remove the precipitate and then placed in the pulse elect