Enterococcus faecalis and application thereof
A technology for Enterococcus faecalis and strains, applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., can solve problems such as low production efficiency, large environmental pollution, and difficulty in purification, and achieve energy saving, production cost reduction, and tyramine capacity. strong effect
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Embodiment 1
[0016] Screening and Preservation of Enterococcus faecalis xltyr001
[0017] Medium:
[0018] MRS liquid medium is: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween 80 1mL, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.5g, manganese sulfate 0.25 g, 1000 mL of distilled water, pH 6.2-6.4, sterilized at 115°C for 15 minutes.
[0019] The lower medium is: peptone 5g, yeast extract 5g, beef extract 5g, sodium chloride 2.5g, glucose 0.5g, Tween 1g, magnesium sulfate 0.4g, manganese sulfate 0.03g, dipotassium hydrogen phosphate 2g, triammonium citrate 2g, calcium carbonate 0.1g, ferrous sulfate 0.04g, vitamin B 1 0.01g, pyridoxal phosphate 0.05g, agar 18 grams. Tyrosine 5 grams, 1000ml distilled water. pH 5.2, autoclaved at 115°C for 15 minutes.
[0020] The upper medium is: bromocresol violet 0.06g, agar 20g, 1000ml distilled water, pH 5.2, sterilized at 121°C for 10min.
[0021] MRS solid medium: ...
Embodiment 2
[0028] Genetic testing of Enterococcus faecalis xltyr001.
[0029] Sources of Primers and Primers
[0030] TD2: 5'-ACATAGTCAACCATRTTGAA-3';
[0031] TD5: 5'-CAAATGGAAGAAGAAGTAGG-3';
[0032] Detection method:
[0033] The 25 μl reaction system includes: GoTaq Green Master Mix 12.5 μl, each primer 1.0 μl, DNA template 2 μl, deionized double distilled water 8.5 μl.
[0034] PCR amplification program: pre-denaturation at 94°C for 5 min, 35 cycles including: denaturation at 94°C for 45 s, annealing at 48°C for 45 s, extension at 72°C for 60 s, final extension at 72°C for 7 min, and cooling to 4°C.
Embodiment 3
[0036] Preparation method of high-concentration tyramide culture solution Pick colonies from the inclined plane and inoculate them into test tubes containing A medium, and culture them at 37°C for 24 hours, and the inoculum size in the test tubes is 10 5 cfu / ml. After repeated activation on the same medium for five times; inoculate the final activated bacterial solution into B medium for static culture at 37°C for 4 days, and the inoculum size is 10 6 cfu / ml; Centrifuge the cultured bacterial solution at 10,000 rpm for 10 minutes, take the supernatant to obtain a high-concentration tyramine solution, and the tyramine content of the high-concentration tyramine solution is 4023.94 μg / ml.
[0037] Substratum: MRS liquid medium (with embodiment 1); Described A medium: the MRS liquid medium of 0.1% tyrosine; Described B medium: 0.005% pyridoxal phosphate+0.1% tyrosine acidic MRS broth.
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