A Bifidobacterium animalis subsp. lactis with the functions of relieving lactose intolerance and reducing triglyceride and its application
A technology for animal bifidobacteria and lactose intolerance, applied in the field of microorganisms, can solve problems such as low triglyceride degradation rate, side effects, sleep problems, etc., achieve excellent acid resistance and bile salt resistance, high strain safety, High safety effect
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Embodiment 1
[0033] Example 1 Screening of Bifidobacterium animalis subsp.lactis HCS04-002CGMCC No.19509 strain
[0034] 1. Primary screening
[0035] Take 500 μL of infant feces samples and add them to 4.5mL TPY synthetic liquid medium for dilution. -6 , take 10 respectively -2 ~10 -6 50 μL of the diluted solution was spread on the synthetic BBL solid medium plate, each gradient was duplicated, and the plate was placed in an anaerobic environment at 39°C for 24-48 hours (the plate was placed in an anaerobic bag).
[0036] 2. Purification
[0037] Pick a single colony with typical characteristics of the target strain (observation shape, size, color, and transparency, etc.), large colony, and strong activity, and carry out streak purification culture on a synthetic BBL solid medium plate, and culture at 37 ° C for 24 ~48h and anaerobic culture at 39°C for 24~48h, repeat this 2~3 times, until the colony characteristics in the streaked plate are consistent;
[0038] The BBL medium: 70g o...
Embodiment 2
[0039]Example 2 Identification of Bifidobacterium animalis subsp.lactis HCS04-002CGMCC No.19509 Strain
[0040] The pure culture separated and purified in Example 1 was further determined to be a pure culture through streaking and smear microscopy, and then the strain identification was carried out, including Gram staining test, contact enzyme test and 16S rDNA full sequence sequencing Identification. The finally identified and isolated strain was a strain of Bifidobacterium animalis subsp. lactis, named Bifidobacterium animalis subsp. General Microbiology Center of Culture Collection Management Committee", the preservation number is CGMCC No.19509.
[0041] The aforementioned Bifidobacterium animalis subsp. lactis HCS04-002 strain is Gram-positive, and the cells are polymorphic rod-shaped and arranged in pairs. Its physical and chemical characteristics are: catalase negative, oxidase negative, can use D-ribose, D-glucose, maltose, lactose, melibiose, sucrose, raffinose.
Embodiment 3
[0042] Example 3 Animal Bifidobacterium animalis subsp.
[0043] The ability of Bifidobacterium animalis subsp. lactis HCS04-002 to withstand the adverse environment of the digestive tract is a prerequisite for its ability to reach the intestinal tract with a high survival rate, survive in the intestinal tract, colonize and grow, and exert its efficacy.
[0044] 1. Acid resistance test
[0045] The three-passage bacterial solution was inoculated into the blank control medium, the basic MRS medium with pH 3.0 and pH 2.0 respectively according to the inoculum amount of 10%. Cultivate statically at 39°C, take samples at 17 hours, perform 10-fold serial dilution with sterilized normal saline, take 1000 μL of bacterial solution with appropriate dilution respectively for mixed bacteria counting operation, repeat twice for each dilution, and culture statically at 39°C Count after 48-72 hours.
[0046] Acid resistance test data indicators:
[0047] The number of viable bacteria (ex...
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