Lactobacillus gasseri LS03 and application thereof
A technology of Lactobacillus gasseri and bacterial suspension, applied in the field of microorganisms, can solve the problems of dependence on imports of probiotic strains, influence on the promotion of probiotics and their products, lack of scientific research evidence for probiotic functions, etc., and achieve good reproductive tract infection. , the prevention or alleviation of reproductive tract infection, the effect of alleviation or prevention of female reproductive tract infection
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Embodiment 1
[0023] Example 1 Lactobacillus gasseri LS03 isolation, identification and preservation
[0024] (1) Separation: The reproductive tract secretions of healthy women of childbearing age were serially diluted, inoculated on anaerobic blood agar medium and MRS solid medium respectively, cultured anaerobically at 37°C for 48 hours, and picked single colonies on the plate to mark Line isolation yielded 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.
[0025] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.
[0026] (3) 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 1...
Embodiment 2
[0033] Example 2 Lactobacillus gasseri LS03 Acid Production Ability Determination
[0034] Lactobacillus gasseri LS03 was inoculated in MRS liquid medium, cultured in an anaerobic workstation at 37°C for 14 hours, and the strain concentration was adjusted to 1×10 6 CFU / mL, inoculated in fresh MRS liquid medium at 5% volume fraction, placed in a full-temperature shaker at 37°C and 150r / min for 0, 5, 10, 15, 20, 24, 28, 32, 36 After 48 hours, the concentration of lactic acid in the fermentation supernatant of the strain was detected by Hitachi automatic biochemical analyzer. At the same time, a pH meter (Sartorius Scientific Instruments (Beijing) Co., Ltd.) was used to measure the pH value of the fermentation broth.
[0035] see results image 3 and Figure 4 . Depend on image 3 It can be seen that Lactobacillus gasseri LS03 started to ferment and produce lactic acid after being inoculated in the medium, and almost reached the saturated concentration after 24 hours, and th...
Embodiment 3
[0036] Preparation of embodiment 3 Lactobacillus gasseri LS03 fermentation supernatant (extracellular secretion), bacterial suspension (thalline)
[0037] Lactobacillus gasseri LS03 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 1×10 7 CFU / mL, centrifuged at 4°C, 6000r / 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 1×10 7 CFU / mL to obtain bacterial suspension (bacteria).
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