Lactobacillus acidophilus for producing exopolysaccharides with anti-tumor, hypoglycemic and lipid-lowering activity and application of lactobacillus acidophilus
A Lactobacillus acidophilus, hypoglycemic and lipid-lowering technology, applied in the field of Lactobacillus acidophilus exopolysaccharide, can solve the problem of separation and purification, and anti-tumor, hypoglycemic and weight loss No patents have been seen, and Lactobacillus acidophilus has not yet been retrieved Patent and other issues, to achieve good gastrointestinal environmental tolerance, reduce postprandial blood sugar levels, and inhibit proliferation
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Embodiment 1
[0082] Embodiment 1: Isolation, purification and identification of bacterial strains
[0083] (1) Isolation and purification of bacterial strains
[0084] Taking yogurt from the Hetao area as a sample, dilute it to 10 times with sterile normal saline -6 , take 10 -4 、10 -5 、10 -6 Spread 100 μl of the three gradient dilutions on the MRS solid plate, incubate at 37°C for 48 hours, select colonies of different shapes and separate them by streaking, until a pure single colony with the same colony shape is obtained, pick a single colony and inoculate it in 5ml of MRS liquid In the culture medium, cultivate overnight, take 800 μl of bacterial liquid and add 200 μl of 60% glycerol to store at -80°C.
[0085] (2) Identification of strains
[0086] The isolated strains were subjected to PCR amplification of 16S rDNA, and the PCR products were sent to Jinweizhi for sequencing. The sequencing results were compared with the nucleic acid sequences in EzBioCloud. The similarity was 91....
Embodiment 2
[0089] Embodiment 2: the growth curve of bacterial strain
[0090] The Lactobacillus acidophilus in Example 1 was inserted onto the MRS solid medium and cultured anaerobically at 37° C. for 48 hours. Pick a single colony in good growth state and inoculate it into MRS liquid medium for activation, then inoculate it into MRS liquid medium with 1% inoculum amount, let it stand for 24 hours, take the bacterial suspension every hour to measure OD600, and draw the philotropic Growth curve of Lactobacillus acidum. Such as figure 1 shown.
Embodiment 3
[0091] Embodiment 3: Determination of acid tolerance of Lactobacillus acidophilus YL01
[0092] The lactic acid bacteria were activated for 3 times, placed in an anaerobic incubator at 37°C, and cultured overnight; the concentration of the bacteria was diluted to 5×10 8 CFU / mL, replace the MRS medium with different pH values, the pH is 2.0, 3.0, 4.0 respectively; put the bacterial solution with different pH in a 96-well plate, set 6 parallel wells for each experimental group, and set up a negative control , and its negative control is MRS with a pH value of 7, which is placed in an anaerobic incubator at 37°C for 4 hours. 20 μL of MTT (5 mg / mL) will be added and placed in an anaerobic incubator for 4 hours. Measure the absorbance value of each hole at 490nm (A experimental group) with an enzyme-linked immunosorbent detector, and compare it with the absorbance value of the negative control group (A control group) to determine the survival rate. The survival rate calculation f...
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