Psychrobacter cryohalolentis, and application thereof in cold storage and preservation of vegetables
A psychrophilic bacillus and salt crystal technology, applied in the field of food preservatives, feed biological additives, and microorganisms, can solve the problem of hindering the mining and understanding of bacteriocin-producing bacteria resources, and it is difficult to perform at temperatures below 10°C or even below 4°C. Antibacterial effect and other problems, to achieve the effect of increasing the scope of application, wide antibacterial spectrum, strong acid and alkali resistance
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Embodiment 1
[0024] Example 1 Screening of Psychrobacterium halocrystals SDWF2-4
[0025] Take the fruit and vegetable samples (temperature 4°C) that have been stored for a long time in the vegetable refrigerated and cold storage of our company without mildew or spoilage, and use sterile water to wash the surface on the ultra-clean workbench, at 10°C, 150rpm / After 20 days of enrichment at 1 min, the LB medium was continuously replenished. After the OD600 value of the bacterial solution reached above 0.5, it was serially diluted and inoculated on a solid medium, cultivated at 10°C for 6 days, and a typical single colony was selected for purification.
[0026] Among them, since the screening target bacteria are unknown, four common screening media in this field were used, namely: LB nutrient medium, lactic acid bacteria medium (Lactic Acid Bacteria Culture Medium, MRS), beef extract peptone medium (Beef Extract Peptone Medium, NA), Columbia Blood Agar Medium (Columbia BloodAgar Medium, CA)....
Embodiment 2
[0035] Example 2 Preparation of Bacteriocins by Fermentation of Psychrobacterium halocrystals SDWF2-4
[0036] Start from the single colony screened and preserved in Example 1, pick a single colony of the bacteria and inoculate it in LB medium, culture it with shaking at 5-25°C for 4-24 hours, centrifuge the culture solution, remove the bacteria, and keep the supernatant , use sodium hydroxide supernatant to adjust the pH value to be consistent with that of distilled water, and then filter through a 0.22 μm filter membrane for later use; put the filtered clear liquid in a beaker, add fully ground ammonium sulfate, and stand for dialysis after reaction , to obtain the crude extract of Psychrobacterium halocrystals SDWF2-4 bacteriocin, dry the crude extract and add different compounding agents to prepare antibacterial agents, which are subpackaged for use.
[0037] The Oxford cup double-layer plate method commonly used in this field was used for screening, and Staphylococcus aur...
Embodiment 3
[0041] The property detection of embodiment 3 bacteriocin
[0042] Carry out following experiment after the bacteriocin crude extract obtained in embodiment 2 is subpackaged:
[0043] (1) thermal stability test
[0044] Put the above-mentioned bulk liquid in water bath at 5°C, 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, and 80°C for 20min, 40min, and 60min respectively. a parallel group. The antibacterial activity of bacteriocin was determined by double-layer plate method with Staphylococcus aureus as indicator bacteria.
[0045] The results show that bacteriocins have certain activity in the range of 5-60°C, but the activity is high or low. Specifically, the activity is the strongest at 20-30°C. After 1 hour, the original activity is basically maintained, which can realize the stable maintenance of antibacterial activity; under the action of 60°C, the effect will be weakened, and it will begin to decline after 20 minutes of treatment, and the decline will be obvious after 1 ...
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