A Lactobacillus acidophilus producing exopolysaccharide with antitumor and hypoglycemic and lipid-lowering activities and its application
A technology of Lactobacillus acidophilus, hypoglycemic and lipid-lowering, applied in the field of Lactobacillus acidophilus of extracellular polysaccharides, can solve the problem that no patents of Lactobacillus acidophilus have been retrieved, separation and purification, anti-tumor, hypoglycemic and weight loss. Patent and other issues, to achieve the effect of inhibiting the proliferation of colon cancer and cervical cancer cells, good gastrointestinal environment tolerance, anti-tumor, hypoglycemic and lipid-lowering activities
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Embodiment 1
[0082] Example 1: Isolation, purification and identification of strains
[0083] (1) Isolation and purification of strains
[0084] Take the yogurt from the Hetao area as a sample and dilute it to 10 times with sterile normal saline. -6 , take 10 -4 , 10 -5 , 10 -6 100 μl of the three gradient dilutions were spread on MRS solid plates, cultured at 37°C for 48 h, and colonies of different shapes were selected for streaking until a pure single colony with the same colony shape was obtained, and a single colony was picked and inoculated into 5 ml of MRS liquid. In the medium, cultured overnight, take 800 μl of bacterial liquid, add 200 μl of 60% glycerol, and 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 in EzBioCloud, and the similarity was 91.29%. One strain of Lactobacillus acidophilus was ...
Embodiment 2
[0089] Example 2: Growth curves of strains
[0090] The Lactobacillus acidophilus in Example 1 was inserted into MRS solid medium and cultured anaerobically at 37°C for 48h. Pick a single colony with good growth status and inoculate it into MRS liquid medium for activation, and then inoculate it in MRS liquid medium with 1% inoculum, let it stand for 24 hours, and measure the OD600 of the bacterial suspension every hour, and draw the Growth curve of Lactobacillus acidophilus. like figure 1 shown.
Embodiment 3
[0091] Example 3: Determination of acid resistance of Lactobacillus acidophilus YL01
[0092] The lactic acid bacteria were activated three 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, replaced with MRS medium with different pH values, pH was 2.0, 3.0, 4.0; bacterial solutions with different pH values were placed in 96-well plates, each experimental group was set with 6 parallel wells, and a negative control needs to be set , and its negative control was MRS with pH 7, which was placed in an anaerobic incubator at 37 °C for 4 h. 20 μL of MTT (5 mg / mL) will be added and placed in an anaerobic incubator for 4 h. The absorbance value of each well at 490nm (A experimental group) was measured in an enzyme-linked immunosorbent assay instrument, and compared with the absorbance value of the negative control group (A control group) to determine the survival rate. The formula for calculat...
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