A strain of Lactobacillus fermentum capable of preventing and/or treating periodontitis and its application
A technology for fermenting lactobacillus and toothpaste, applied in the field of microorganisms, can solve the problems of high cost, increase production cost, cannot inhibit the growth of Porphyromonas gingivalis, etc., and achieve the effect of relieving periodontitis and having a huge application prospect.
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preparation example Construction
[0064] The method of preparing the bacterial suspension involved in the following examples is as follows:
[0065] With the suspension of the nucleus, the Nucleus Bacillus is inoculated into the BHI medium in the inoculation amount of 2% (v / v) in the total volume of BHI medium, and then at 37 ° C for anaerobic culture for 48 h, with BHI Medium-based modifier concentration is 1 × 10 9 CFU / ml.
[0066] Gubprinocensis Suspension: Inoculation of the gingival porphty cells inocatalyst in the BHI medium in the amount of 2% (v / v) of the total volume of BHI medium, 48 h at anaerobic culture at 37 ° C After using BHI medium, the hydrochloride concentration is 1 × 10 9 CFU / ml.
[0067] Intermediate Placeonius suspension: Intermediate P. aeruginus is inoculated into BHI medium in a inoculation amount of 2% (V / V) in BHI medium, and in anaerobic culture at 37 ° C for 48 hours. Based sized hydrochloride concentration is 1 × 10 9 CFU / ml.
[0068] The method of equipped with the PBS b...
Embodiment 1
[0070] Example 1: Screening and strain identification of Lactobacillus Fermentum CCFM1139
[0071] 1, screening
[0072] Since the feces sample from the healthy person from Yangzhou area, after pretreatment, the sample is preserved in the -80 ° C refrigerator, remove the thaw, mix the sample, and absorb 0.5ml sample to 4.5ml 9g / L Physiological saline was diluted, selecting a suitable gradient diluent coated on MRS medium containing 20 g / L agar 48 h at 37 ° C for 48 h, and a typical colonies of fermentation Lactobacillus were selected to a MRS medium containing 20 g / L agar. Undiren purification, picking up a single colony transfer to MRS medium, 30% glycerol deposit, resulting in strain CCFM1139 and synchronous strains 1 to 19; wherein the typical colonies of the fermented Lactobacillus are small white and translucent. Round.
[0073] 2, identification
[0074] The genome of strain CCFM1139 and strains 1 to 19 was extracted, and 16S RDNA of strain CCFM1139 and strain 1 to 19 ...
Embodiment 2
[0075] Example 2: The effect of Lactobacillus Fermentum CCFM1139 on the amount of mixed biofilm is divided into two groups, which are fermented Lactobacillus and blank control groups;
[0076] Among them, the fermentation Lactobacillus system is handled in accordance with the following method:
[0077] (1) The fermented Lactobacillus CCFM1139, a bacterium, fermented Lactobacillus 1 to 19, respectively, inoculated into the MRS medium in the MRS medium total volume of 2%, at 37 ° C for 24 h, obtained culture solution, will be cultured The fluids were centrifuged at 8000 r / min, 4 ° C for 5 min, and the supernatant was obtained, and the supernatant was filtered at 0.22 μm sterile filmers, respectively, respectively. 9 CFU / mL of the fermentation of Lactobacillus CCFM1139, the supernatant of the fermentation of Lactobacillus 1 to 19;
[0078] (2) In the 96-well plate, 40 μl of the gingival porphty cells and the nucleus of the nucleus, and the mixed bacteria suspension is obtained, a...
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