Lactobacillus reuteri SF-L-25 with helicobacter pylori inhibiting function and application thereof
A technology of SF-L-25 and Lactobacillus reuteri, applied in the field of microorganisms, can solve problems such as gastrointestinal flora imbalance, antibiotic gastrointestinal dysfunction, etc., achieve high safety, wide application value, and inhibit Helicobacter pylori The effect of growth
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Embodiment 1
[0040] Example 1 Screening of Lactobacillus reuteri SF-L-25
[0041] ① Source of bacteria: healthy human gastric juice
[0042] ② Plate medium: beef extract 10.0g, yeast extract 5.0g, peptone 10.0g, grape 20.0g, K 2 HPO 4 2.0g, Sodium acetate 5.0g, MgSO 4 ·7H 2 O 0.2g, MnSO 4 ·4H 2 O0.05g, Tween 801.0g, triammonium citrate 2.0g, agar 15.0g, add distilled water to 1000ml.
[0043] ③Isolation of strains
[0044] The sample liquid was diluted ten-fold, and 100 μL of each gradient was evenly spread on the MRS solid plate medium, then taken out and placed in an anaerobic tank, and cultured overnight at 37°C. The next day, colonies of different shapes and sizes were found on the plate, and some characteristic colonies of lactic acid bacteria were picked for the second-generation streak purification treatment, and the plate was anaerobic activated for 1 day to obtain a culture, followed by strain identification.
[0045] ④ Determination of 16S rRNA sequence
[0046] The 16S ...
Embodiment 2
[0049] Embodiment 2 Determination of antibacterial ability of Lactobacillus reuteri SF-L-25
[0050] Lactobacillus reuteri SF-L-25 was activated for 2 generations, and the bacterial suspension was filtered through a 0.22 μm sterile filter to obtain the Lactobacillus reuteri SF-L-25 fermentation supernatant. The H.pylori bacterial suspension of activated 2 generations was adjusted to 10 8 CFU / mL, 100 μL was spread on Columbia blood plate, 150 μL Lactobacillus reuteri SF-L-25 fermentation supernatant was added to the Oxford cup, and the control group was added with the same amount of distilled water, and was cultured in a three-gas incubator 72h, determine the diameter of the inhibition zone (see figure 1 ). Do 1 control and 3 parallel experiments.
[0051] Table 1 Bacteriostatic ability of Lactobacillus reuteri SF-L-25
[0052] group Inhibition zone / (mm) control 0 Test group 1 12.92 Test group 2 12.38 Test group 3 12.15
[0053] The ...
Embodiment 3
[0054] Example 3 Determination of Adhesion Ability
[0055] Human gastric adenocarcinoma cells (AGS) were seeded in 96-well plates (2×10 4 Cells / well) were cultured overnight, and after the cells adhered to the wall, F-12 resuspended Helicobacter pylori and Lactobacillus reuteri SF-L-25 were added at a multiplicity of infection of 100 for 2 hours, respectively, and then 200 μL of urease reagent was added for 3 hours. Measure the absorbance at 550nm wavelength, and do 1 control and 3 parallel experiments.
[0056] Table 2 Determination of the ability of Lactobacillus reuteri SF-L-25 to inhibit the adhesion of Helicobacter pylori
[0057] group Adhesion rate / (%) control 100 Test group 1 69.62 Test group 2 70.38 Test group 3 69.25
[0058] The results showed that the adhesion rate of Helicobacter pylori to AGS decreased significantly after treatment with Lactobacillus reuteri SF-L-25.
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