Seabuckthorn seed polypeptide with antibacterial effect and preparation method and application thereof
A technology of bacteriostatic effect and sea buckthorn seeds is applied in the field of preparation of sea buckthorn polypeptides, which can solve the problems of interfering with bacteriostatic effect and the like
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Embodiment 1
[0018] Take 20g of crushed defatted seabuckthorn seed dregs (30% protein content), add 300ml of distilled water, place it at 25°C for 8 hours, adjust the pH to 6.5, add 0.18g of ProteAX protease and hydrolyze it at 50°C for 3h; put it into a glass bottle During the process, tighten the lid, inactivate the enzyme with a 1000W household microwave oven for 5 minutes, centrifuge the enzymolysis solution at 4000r / min to obtain the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and concentrate the filtrate until the polypeptide content reaches 2 %save.
[0019] The above concentrated solution was diluted to 20mg / ml for antibacterial test, the bacteria used were Staphylococcus aureus and Salmonella, and the culture medium was nutrient agar medium. When the medium is boiled and cooled to 50-60°C, it is poured into a petri dish, and after solidification, Staphylococcus aureus and Salmonella are respectively inoculated on the surface of the m...
Embodiment 2
[0021] Take 50g of crushed defatted seabuckthorn seed dregs (30% protein content), add 1000ml of distilled water, place it at 25°C for 10 hours, adjust the pH to 7.0, add 0.45g of ProteAX protease, hydrolyze it at 46°C for 4.5h, put it in a sealed In a glass bottle, use a 1000W household microwave oven to inactivate the enzyme for 7 minutes, centrifuge the enzymolysis solution at 4000r / min to take the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and concentrate the filtrate until the polypeptide content reaches 16 %.
[0022] Dilute the concentrated solution to 5, 10, 20, 40, 80, 160 mg / ml for antibacterial test, the bacteria used are Salmonella, Staphylococcus aureus and Pseudomonas aeruginosa, and the culture medium is nutrient agar medium. Cultivate and observe method with described in embodiment 1, obtain concentration inhibition curve ( image 3 ). It can be seen from the concentration inhibition curve that the optimal poly...
Embodiment 3
[0024] Take 100g of pulverized defatted seabuckthorn seed dregs (30% protein content), add 1500ml of distilled water, place it at 25°C for 8 hours, adjust the pH to 7.0, add 0.63g of ProteAX protease, hydrolyze it at 55°C for 1.5h, put it into a sealed container In a glass bottle, use a 1200W household microwave oven to heat and inactivate the enzyme for 10 minutes, centrifuge the enzymolysis solution at 4000r / min to get the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and take the filtrate and then use an ultrafiltration filter with a cut-off pore size of 4000Da. Membrane ultrafiltration, take the concentrated solution to separate short and medium molecular weight polypeptides of 4000 to 5000 Da, and take the filtrate and use an ultrafiltration membrane with a cut-off pore size of 2000 Da for ultrafiltration, and take the filtrate to obtain small molecular peptides below 2000 Da. Peptides of different molecular weights were concen...
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