Human-derived lactobacillus acidophilus resisting gastrointestinal tract stress and application thereof
A technology of Lactobacillus acidophilus and gastrointestinal stress, applied in the direction of Lactobacillus, application, bacteria used in food preparation, etc., can solve the problems of large differences in probiotic activity, achieve good inhibitory effect, rich nutrition, strong The effect of adhesion properties
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Embodiment 1
[0019] Example 1: Screening and identification of bacterial strains.
[0020] Bacteria and hosts in the microecosystem have a corresponding supply and demand relationship, and the adhesion of some probiotics is specific to the host species; the types of bacteria in the microecosystem in the intestinal tract of infants are less than those of adults, but the probiotics The types and numbers of bacteria are more than those of adults; with the development of infants, the intestinal morphology will also change accordingly, coupled with changes in diet after weaning, the intestinal flora of infants will undergo a huge change: facultative anaerobic The number of bacteria decreased, the number of strictly anaerobic bacteria such as Bacteroides gradually increased and became the dominant flora, and the types of intestinal flora increased. At 1 year old, the diversity of intestinal flora was close to the characteristics of adult intestinal flora. Therefore, in order to exert a stronger ...
Embodiment 2
[0049] Example 2: Research on anti-gastrointestinal stress performance.
[0050] By simulating the human digestive tract environment, the survival ability of Lactobacillus acidophilus NCU402 in the gastrointestinal tract was explored, so as to provide a reference for whether this strain of lactic acid bacteria can play a health-promoting role in the human digestive tract environment.
[0051] 2.1 Determination of resistance to high osmotic pressure environment.
[0052] NaCl was added to the MRS liquid medium to make the mass fractions 0%, 2%, 4%, 6%, 8% and 10%, respectively, and sterilized to prepare MRS liquid medium with different osmotic pressures. The activated NCU402 was inserted into MRS liquid medium with different NaCl concentrations, cultured at 37°C for 24 hours, and then sampled to measure the concentration of viable bacteria; the curves were drawn under MRS liquid medium with different osmotic pressures.
[0053] The results of NCU402 cultured in MRS liquid medi...
Embodiment 3
[0063] Example 3: Determination of the adhesion ability of Caco-2 colon cancer cells.
[0064] NCU402 has strong acid and bile salt resistance, but the strength of its adhesion to human digestive tract cells is the key first step for Lactobacillus acidophilus to exert its probiotic function. Currently, three types of intestinal cell systems are used in in vitro cell adhesion research: HT-29 colonic cell line, HT29-MTX cell line and Caco-2 cell line.
[0065] Detection by Gram staining method: Digest Caco-2 cells in glass culture flask with 0.25% EDTA-trypsin digestion solution, centrifuge at 1000rpm for 5min, carefully remove supernatant, add quantitative sterile PBS buffer and mix well, place Count on a hemocytometer; centrifuge again at 1000 rpm for 5 min and add cell culture medium to adjust the cell concentration to 1.0×10 4 cell / mL, spread on a 6-well culture plate at 2 mL per well, at 37°C, 5% CO 2 , 95% relative humidity and cultivated until the cells grow into a dens...
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