A kind of extraction method of antibacterial protein of bow crab and its application in food preservation
A technology of antibacterial protein and extraction method, which is applied in the direction of food preservation, food ingredients as antimicrobial preservation, application, etc., can solve the problems of low price and impact, improve activity and purity, broaden application fields, and benefit The effect of antibacterial activity
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Embodiment 1
[0038] Preparation of antimicrobial protein extracts
[0039] The flow chart of the antibacterial protein extraction process of the Chinese crab figure 1 As shown, it mainly includes the following steps:
[0040] (1) Preparation of flat crab tissue: take the whole flat crab, only take its gills and intestinal organs and other tissues, discard the limbs, shell and other limbs, and obtain the flat crab tissue.
[0041] (2) Preparation of flat crab tissue homogenate: the ratio of flat crab tissue and distilled water is such that 2 mL of distilled water corresponds to 1 g of flat crab tissue homogenate. The homogenate was pulverized with a grinder to obtain a flat crab tissue homogenate. The appearance of the obtained tissue homogenate is as follows: image 3 (a) shown.
[0042] (3) Preparation of Antibacterial Protein Crude Extract 1: Centrifuge the flat crab tissue homogenate at a speed of 12000 rpm for 10-15 minutes, and collect the supernatant. However, since there is a cer...
Embodiment 2
[0049] Antibacterial Activity Determination Index Evaluation
[0050] The sample solution in this embodiment is the antibacterial protein extract in the different extraction steps of Example 1.
[0051] Strain activation: Take a sterile Erlenmeyer flask and add 30 mL of liquid LB medium; take out the preserved strains from the -80°C refrigerator, draw 500 μL of E. Store in a 4°C refrigerator for later use; the activation method for Staphylococcus aureus is the same as above, and place it in a 4°C refrigerator for later use after culturing for 6 hours.
[0052] Determination of the concentration of the bacterial solution: use the plate dilution method to determine the concentration of the bacterial solution. The specific method is as follows: 9 EP tubes are numbered, and 900 μL of sterile water is added to each; Mix well, pipette 100 μL into tube 2, and add to tube 9 in turn; take 100 μL from tubes 7, 8, and 9 and spread them on LB medium plates, with 3 parallel plates in each...
Embodiment 3
[0056] Activity comparison of other 4 kinds of marine biological source extracts and other 4 kinds of crab extracts
[0057] Extracts from other 4 kinds of marine organisms: scallops, oysters, river clams and mussels were purchased from the market, and these 4 kinds of organisms were respectively processed according to the extraction step 1 of the above-mentioned flat crab crude extract. The extract obtained from each organism was passed through a 0.22 μm sterile filter membrane and stored in a sterile EP tube for later use; the antibacterial activity was determined as described in Example 2 above.
[0058] Activity comparison of extracts from other species of crabs: hairy crabs, blue crabs, red star portunus and portunus lanceolata were purchased from the market, and these four crabs were processed according to the extraction steps of the crude extracts of flat crabs mentioned above. For Scylla serrata whose crude extract has antibacterial activity against Staphylococcus aure...
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