A kind of preparation method and application of antibacterial polypeptide of sand jujube seed
A technology of antibacterial polypeptides and seeds, which is applied in the preparation methods of peptides, antibacterial drugs, antifungal agents, etc., can solve the problem of too much seed protein
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Embodiment 1
[0015] a. Wash 300g of Elaeagnus radix fruit with water, clean and dry the seeds naturally, pulverize them, and degrease with petroleum ether to obtain 48g of degreasing Elaeoptera erae seeds;
[0016] b. With the defatted seeds of Elaeagnus eraeae obtained in step a, at room temperature, use 10% acetic acid with a concentration of 10% acetic acid to extract the total polypeptide of the seeds of Elaeoptera eraeaceae under stirring at a ratio of w / v=1:7 at a temperature of 20° C. for a period of time After 2 hours, filter the supernatant, then pour the supernatant into anhydrous acetone for overnight precipitation according to v / v=1:5, centrifuge at 4°C and 12000r / min for 3 times, each time for 15min, and Each precipitate was washed with cooled anhydrous acetone, and then the precipitate was naturally dried to obtain the total polypeptide extract. The total protein content was measured by the biuret method, and the total protein content was 72.6% by ultraviolet detection at 540n...
Embodiment 2
[0019] Use the agar plate method to detect the antibacterial effect of the sample on the three bacteria, and measure the size of the antibacterial ring of the sample on the three bacteria, so as to judge the antibacterial ability of the sample:
[0020] Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia coli were cultured overnight in LB liquid medium at 175rpm and 37°C until the OD600 was around 2.0 (1×106CFU / mL). Mix 10 μL of solution with 8 mL of LB medium containing 0.7% agar and spread it on the LB medium containing 25 mL of 1.5% agar. , cultivate overnight at 37°C, and measure the diameter of the inhibition zone. Ampicillin was used as a positive control, and normal saline was used as a negative control.
[0021] Table 1 Antibacterial results of antibacterial peptides in seeds of Eleuthero jujube (M±m)
[0022]
Embodiment 3
[0024] The tandem mass spectrometer is composed of two mass spectrometers connected in series. The first mass analyzer pre-separates or adds energy modification to the ions, and the second mass analyzer analyzes the results; The crude peptides extracted from the seeds were analyzed by mass spectrometry, and four peptide molecules with molecular weights of 5492.9Da, 6551.5Da, 4400.5Da and 2837Da were obtained by mass spectrometry.
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