Lactobacillus plantarum and application of lactobacillus plantarum in preparation of medicines for treating infectious vaginosis
A technology of Lactobacillus plantarum and vaginal pathogenic bacteria, which is applied in the field of microorganisms, can solve the problems of poor antibacterial effect, no patent publications have been found, gastrointestinal intolerance, etc., and achieve good antibacterial ability and good acid production Effect of hydrogen peroxide capacity, good acid resistance and bile salt tolerance
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Embodiment 1
[0046] Embodiment 1: Screening and identification of strains
[0047] (1) Isolation and purification of bacterial strains
[0048] The Lactobacillus plantarum 021 was screened in mung bean physalis produced in Henan area. The mung bean milk solution was diluted with sterile PBS buffer solution and spread on the MRS plate medium, cultured at 37°C for 48 hours under anaerobic conditions, and well-separated single colonies were picked and streaked. A single colony was picked by streaking multiple times, and the purified strain was stained with Gram staining, and the morphology and distribution of the bacteria were observed under a microscope. This strain is a Gram-positive round coccus, see figure 1 .
[0049] The bacterial solution was diluted 10 times with sterile saline to 10 -6 , take 10 -4 、10 -5 、10 -6100 μl of the three gradient dilutions were spread on the MRS broth solid plate, cultured at 37°C for 48 hours, and colonies of different shapes were selected for strea...
Embodiment 2
[0054] Embodiment 2: Lactobacillus plantarum 021 acid tolerance
[0055] (1) After activating the lactic acid bacteria for 2-3 times, use 1.0X10 9 The concentration of CFU / mL was respectively inoculated in MRS broth medium with pH 2, 3 and 4 according to 1% inoculum volume and cultured for 4 hours.
[0056] (2) After the culture is over, use the normal MRS broth medium as a control, mix the bacterial solution thoroughly, add it to a 96-well plate that has been sterilized in advance, 100 μL per well, set 3-6 duplicate wells, and use multiple The functional microplate reader measures the absorbance at 600nm to detect the inhibition rate of the culture medium with different pH values on lactic acid bacteria.
[0057] Formula: survival rate (%)=(treatment group / control group) X 100%
[0058] The result is as figure 2 Shown: Through experiments, it was found that the survival rate of Lactobacillus plantarum 021 in the MRS broth medium with pH 2 and pH 3 is relatively high, wh...
Embodiment 3
[0059] Embodiment 3: Lactobacillus plantarum 021 tolerance to bile salts
[0060] (1) After activating the lactic acid bacteria for 2-3 times, use 1.0X10 9 The concentration of CFU / mL was respectively inoculated in MRS broth medium containing 0.1%, 0.2%, and 0.3% porcine bile salt according to the inoculum size of 1%, and cultured for 4 hours.
[0061] (2) After the culture is over, use the normal MRS medium as a control, mix the bacterial solution thoroughly, add it to a 96-well plate that has been sterilized in advance, 100 μL per well, set 3-6 duplicate wells, and use a multifunctional enzyme The absorbance at 600nm was measured by a standard instrument to detect the inhibition rate of different contents of bile salts on lactic acid bacteria.
[0062] Formula: survival rate (%)=(treatment group / control group) X 100%
[0063] The result is as image 3 Shown: Through experiments, it is found that Lactobacillus plantarum 021 can grow in MRS broth medium containing different...
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