Lactobacillus paracasei separated from Tibetan ali yak milk lumps and application of lactobacillus paracasei
A technology of Lactobacillus and para-cheese, applied in the field of microorganisms, can solve the problems of poor inhibition of pathogenic bacteria and other problems
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Embodiment 1
[0034] The screening of embodiment 1 bacterial strain
[0035] Take a sample of yak milk pimple in the Ali area of Tibet, and first use the selective medium MRS and BL to conduct a preliminary screening of the bacteria: Weigh an appropriate amount of sample and dissolve it in normal saline to 10ml. Shake the activation solution evenly, put 0.1mL of the activation solution on the selection medium, dilute and spread it, incubate at 37°C for 48 hours, observe the morphological characteristics of the colony, pick a single colony, and continue to separate and purify until pure colonies can be seen under the microscope. Subsequent addition of 0.2% CaCO to the selection medium 3 Re-screening: the pure colonies obtained from the primary screening were streaked and cultured in the re-screening medium for 24h-48h, after the end of the culture, observe whether there is a transparent circle, and select a single colony with a large transparent circle for Gram staining. The strain that w...
Embodiment 2
[0037] Example 2 Molecular biology identification of 16S rRNA
[0038] Using high-fidelity enzymes, 16SrDNA sequencing and strain identification were performed on the target strain obtained in Example 1, and a phylogenetic tree was drawn. The primers used were: forward primer 27F: 5'-AGAGTTTGATCATGGCTCAG-3'; reverse primer 1492R: 5'-ACGGCTACCTTGTTACGACTT-3'. The PCR reaction system is: 25μl Mix; 1μl sample; 20μl ddH 2 O; 2μl 27F; 2μl 1492R; PCR reaction temperature: 98°C, 2min; 98°C, 10s, 58°C, 10s, 72°C, 7s, 35x; 72°C, 2min; store at 4°C. The obtained PCR products were sequenced, and then a phylogenetic tree was constructed using MEGA8.0. Phylogenetic tree such as image 3 . According to the phylogenetic tree, it can be known that the strain is Lactobacillus paracasei.
[0039] The measured 16S rRNA gene sequence of Lactobacillus paracasei is shown in SEQ ID NO:1.
[0040] SEQ ID NO: 1:
[0041]
[0042]
[0043] The purpose bacterial strain obtained is named Lac...
Embodiment 3
[0044] Embodiment 3 Lactobacillus paracasei AL1 Plateau LPA-1 bacterial strain is to the inhibition of Salmonella
[0045] The activated Lactobacillus paracasei AL1 Plateau LPA-1 strain was inserted into the MRS liquid fermentation broth with a 5% (v / v) inoculation ratio, cultivated at 37°C, and the fermentation broth was cultivated for 48 hours, and the fermentation broth fermented for 48 hours was centrifuged at 1000rpm for 10min Take the supernatant, and filter it through a 0.45 μm filter membrane to obtain a fermentation treatment liquid.
[0046]The 16S rRNA gene sequence preserved in the glycerol tube is as shown in SEQ ID NO: 2. The Salmonella is first activated, second-generation activated, and seed liquid prepared on the BS medium, and the seed liquid of 100 μl of Salmonella (1×10 8 CFU / ml) on the BS medium, use a sterilized and dried Oxford cup to stand on the surface, add 100 μl of fermentation treatment solution to well A; add 100 μl of sterilized MRS solution to w...
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