Lysozyme has the characteristics of an enzyme and is widely used for food preservation, and is a raw material of a bacteriostatic agent and an antibacterial agent. The use of egg white to extract lysozyme has relatively low production investment, simple processing methods, and wide marketability. It is an effective way for rural development of commodity production and development of egg resources.
First, the preparation of egg white, resin, phosphoric acid, sodium hydroxide, hydrochloric acid, sodium chloride, anhydrous acetone, phosphorus pentoxide as raw materials. Take 70 kg of fresh or frozen egg white, adjust the pH to about 8, and filter with a copper sieve to remove the umbilical cord and other substances in the egg white.
Second, adsorption plus cold water to cool and stir the egg white, until the temperature dropped to 5 degrees, slowly add 724-type resin 10 kg, the temperature is controlled at 5 degrees -10 °C, and then continue stirring for 5 hours, egg white liquid into the cold room, at 0 degrees - 10 degrees overnight.
3. Elution The upper egg white is poured out and the remaining resin is washed with water to remove attached proteins. Wash a total of 4 times, and finally filter dry water. The resin was added to the resin three times with 24 kg per liter of 0.15 molar phosphate buffer. After stirring for 15 minutes each time, the water was removed by suction filtration. The enzyme was then eluted with 18 kg of 10% aqueous ammonium sulfate solution. The eluent was combined and solid ammonium sulfate corresponding to 32% of the eluent was added to precipitate a white precipitate.
4. Dialysis The precipitate was dissolved in 1.5 times the volume of distilled water and dialyzed in a refrigerator for 48 hours to remove most of the ammonium sulfate.
5. Salting-out The filtered solution is filtered to obtain a clarified dialyzate. A sodium hydroxide aqueous solution is added with a dropper and the pH is carefully adjusted to 8.5-9. If a white precipitate appears, it should be immediately centrifuged. Then, 3 mol of hydrochloric acid per liter was added to the mixer, the pH was adjusted to 3-4, and then a volume of 5% solid sodium chloride was added to produce a white precipitate. After 8 hours of storage in a 0°-10° cold room, the lysozyme precipitate was filtered by suction.
6. Drying Add 10 times the amount of anhydrous acetone to the precipitate, and stir the precipitate to make it loose. Put it in the cooling place for several hours, filter off the acetone, obtain the precipitate, and then use phosponpentane pentoxide to dry it to obtain the lysozyme. Powder raw materials.
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