A common Bacteroides strain capable of protecting intestinal permeability and its application
A technology of Bacteroides vulgaris, deposit number, applied in the direction of application, Lactobacillus, bacteria used in food preparation, etc.
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Embodiment 1
[0040] Example 1: Isolation and screening of Bacteroides vulgaris CCFM1152
[0041] 1. Sample collection
[0042] A fecal sample was collected from a 50-year-old middle-aged man in Dieqiao Apartment, Hedong District, Tianjin. The sample was placed in a stool tube filled with 30% glycerol, stored in a thermal insulation box with an ice pack, and quickly placed in the laboratory after being brought back to the laboratory - 80 ℃ refrigerator to be separated and screened.
[0043] 2. Isolation and purification of lactic acid bacteria
[0044] (1) Dilution coating: take about 0.5 g of the content stored in 30% glycerol and add it to a 10 mL centrifuge tube containing 4.5 mL of physiological saline under a sterile environment to obtain 10 -1Diluent, repeat the above dilution steps to obtain 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 Diluent;
[0045] (2) Coating culture: Pipette 100 μL of the above 10 -4 , 10 -5 , 10 -6 The three gradient dilutions were separated and screened ...
Embodiment 2
[0060] Example 2: Effect of Bacteroides vulgaris CCFM1152 on mouse intestinal fluorescein-labeled dextran (FITC)
[0061] 1. Preparation of Bacteroides vulgaris CCFM1152 intragastric fluid
[0062] The Bacteroides vulgaris CCFM1152 obtained in step 2 of Example 1 was streaked in BHI solid medium, and cultivated under anaerobic conditions at 37°C for 48 hours to obtain a single colony; the single colony on the BHI solid medium was picked and inoculated into BHI liquid medium medium, enrichment culture under anaerobic conditions at 37°C for 18-20 hours for activation, and continuous activation for two generations to obtain an activated culture; the activated culture is inoculated into BHI liquid culture at an inoculum amount of 2% to 5% (v / v). In the base, enriched and cultured under anaerobic conditions at 37°C for 18 to 20 hours to obtain bacterial liquid, centrifuged at 8000 g for 10 min to obtain bacterial sludge, washed with sterile saline for 3 times, and then collected th...
Embodiment 3
[0079] Example 3: Application of Bacteroides vulgaris CCFM1152
[0080] Common Bacteroides CCFM1152 can be used to prepare milk, and the specific preparation process of milk is as follows:
[0081] (1) The secondary purified culture solution of Bacteroides vulgaris CCFM1152 obtained in Example 1 was inoculated into the culture medium with an inoculum of 3% (v / v), and cultured at a temperature of 37° C. for 18 hours to obtain a bacterial solution; The liquid was centrifuged to obtain bacterial slurry; the bacterial slurry was washed three times with pH 7.2 phosphate buffer, and then resuspended with a protective agent to a concentration of 1 × 10 10 CFU / mL to obtain a suspension; pre-incubate the suspension for 60 min at a temperature of 37°C and freeze-dry to obtain a bacterial agent;
[0082] Wherein, the preparation method of the culture medium is: adding cysteine hydrochloride 1g / L, hemin 0.01g / L and vitamin K1 0.002g / L to the brain heart infusion, and the pH is 7.0;
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