Lactobacillus fermentum TY-G04 with bowel relaxing effect and application of lactobacillus fermentum TY-G04
A technology of Lactobacillus fermentum and laxative, applied in the field of microorganisms, can solve the problems of difficult treatment, easy repetition, easy recurrence, etc., and achieve the effect of good digestive tract resistance and intestinal colonization ability
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Embodiment 1
[0049] Isolation, purification and identification of TY-G04
[0050] 1. Experimental materials
[0051] Collect naturally fermented yak yoghurt from the herdsman's house in Hongyuan County, Aba Tibetan and Qiang Autonomous Prefecture, Sichuan Province: take the yak yoghurt made by the herdsman's house with a sterile spoon, and put it into 15mL sterile spinner containing appropriate amount of sterile calcium carbonate and soluble starch. Cover the centrifuge tube (calcium carbonate: soluble starch ratio is 1: 1), screw the cap tightly after stirring evenly, then put it into a refrigerator, and transport it back to the laboratory for the purification and separation of lactic acid bacteria immediately.
[0052] 2. Separation and purification of TY-G04
[0053] Under sterile conditions, pipette 1mL sample into 9mL sterile normal saline, vortex to mix, that is 10 -1 sample diluent, and then serially diluted 10 times to 10 -7 , select 10 -5 、10 -6 、10 -7 100 μL of the diluted ...
Embodiment 2
[0064] Determination of TY-G04 Gastrointestinal Fluid Tolerance and Cell Adhesion
[0065] 1. Experimental materials
[0066] Lactobacillus fermentum TY-G04 with laxative effect: isolated from naturally fermented yak yoghurt from herdsmen in Hongyuan, Sichuan, and preserved in the General Microbiology Center of China Committee for the Collection of Microbial Cultures, numbered CGMCC No.23752.
[0067] 2. Determination of TY-G04 tolerance to pH3.0 artificial gastric juice
[0068] TY-G04 was inoculated in MRS liquid medium at 2% inoculation amount, cultured at 37°C for 18 hours, and 10 mL was centrifuged at 4000 r / min for 10 minutes to collect bacterial precipitates, washed twice with sterile normal saline, and resuspended In an equal volume of sterile saline, it is a bacterial suspension. Mix the prepared bacterial suspension with artificial gastric juice (0.2% NaCl, 0.35% pepsin 1:10000, adjust pH=3.0 with 1mol / L HCl, filter and sterilize for later use) in a ratio of 1:9, a...
Embodiment 3
[0083]Improvement effect of TY-G04 on constipation
[0084] 1. Experimental materials
[0085] Lactobacillus fermentum TY-G04 with laxative effect: isolated from naturally fermented yak yoghurt from herdsmen in Hongyuan, Sichuan, and preserved in the General Microbiology Center of China Committee for the Collection of Microbial Cultures, numbered CGMCC No.23752.
[0086] 2. Animal verification experiments
[0087] 2.1. Grouping and handling of experimental animals
[0088] Select 7-week-old BALB / c male mice (20g-25g) and raise them in a standardized laboratory at room temperature 25±2°C, relative humidity 50±5%, 12h light / 12h dark, and start the experiment after one week of adaptive feeding . After the adaptation period, the mice were randomly divided into a normal group, a constipation group, and a TY-G04 group, with 10 mice in each group.
[0089] The experimental period was 18 days, and 1-14 days was the TY-G04 intervention period. The normal group and the constipation ...
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