Lactobacillus gasseri LGV03 and application thereof
A technology of Lactobacillus gasseri and bacterial suspension, applied in the field of microorganisms, can solve problems such as affecting the promotion of probiotics and their products, lack of scientific research evidence for probiotic functions, and dependence on imports of probiotic strains, and achieve good prevention of allergic reactions. , inhibition of mast cell degranulation
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Embodiment 1
[0028] Example 1 Isolation and identification of Lactobacillus gasseri LGV03
[0029] (1) Separation: After serially diluting the secretions of the healthy female reproductive tract, they were respectively inoculated in TPY solid medium, MRS solid medium, BDS solid medium, and BS solid medium, cultured anaerobically at 37°C for 48 hours, and picked Single colonies on the plate were streaked to obtain pure colonies. The pure colony on the plate was inoculated into MRS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.
[0030] (2) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR using 16S rDNA universal primers 27F and 1492R, followed by sequencing to identify the species of the strain. The sequences of the universal primers 27F and 1492R are as follows:
[0031] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; SEQ ID ...
Embodiment 2
[0049] Preparation of embodiment 2 Lactobacillus gasseri LGV03 fermentation supernatant (extracellular secretion), bacterial suspension (thalline)
[0050] Lactobacillus gasseri LGV03 was activated and cultured and inoculated in MRS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 2×10 8 CFU / mL, centrifuged at 4°C, 8000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 2×10 8 CFU / mL to obtain bacterial suspension (bacteria).
Embodiment 3
[0051] Example 3 Effect of Lactobacillus gasseri LGV03 on Compound 48 / 80 stimulating zebrafish to secrete Tryptase
[0052] Sodium cromoglycate and Na-benzoyl-DL-arginine-p-nitroamide hydrochloride (BAPNA) were purchased from Shanghai Yuanye Biotechnology Co., Ltd., and Compound 48 / 80 was purchased from Sigma-Aldrich.
[0053] The healthy wild-type AB line zebrafish developed to 5dpf (days post fertilization) was selected and placed in a 96-well cell culture plate, 10 / well. The experiment set up normal group, model group, intervention group (positive control group, bacterial suspension group, fermentation supernatant group) and solvent zeroing group, and each group set up 6 replicate wells. The normal group was added with PBS, the model group was added with PBS, the positive control group was added with cromolyn sodium solution (100 μg / mL), the bacterial suspension group was added with bacterial suspension, the fermentation supernatant group was added with fermentation superna...
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