Lactobacillus fermentum KP101 and application thereof
A technology of Lactobacillus fermentum, KP101, applied in the directions of Lactobacillus, application, Streptococcus/Lactococcus, etc., can solve the problems of influence, reduce TC and LDL-C levels, etc., to improve HDL-C level, anti-pathogenic bacteria Good effect on infection, tolerance to bile salt stress
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Embodiment 1
[0027] Example 1 Screening and identification of Lactobacillus fermentum KP101
[0028] 1. Screening of bacterial strains
[0029] The kimchi samples purchased in the market were isolated from lactic acid bacteria in the laboratory; -1 Gram-positive strains were selected by observing the colony morphology with Gram staining method; The selected strains were then cultured in liquid MRS medium at 37°C for 24 hours, inoculated according to the inoculation amount of 3% of the culture medium, and activated and passaged continuously for three times to ensure the vigorous growth of the bacteria;
[0030] Liquid MRS medium contains peptone 10.0g, beef extract 10.0g, yeast extract 5.0g, diammonium hydrogen citrate [(NH4)2HC6H5O7] 2.0g, glucose (C6H12O6 H2O) 20.0g, Tween 80 1.0mL, sodium acetate (CH3COONa·3H2O) 5.0g, dipotassium hydrogen phosphate (K2HPO4·3H2O) 2.0g, magnesium sulfate (MgSO4·7H2O) 0.58g, manganese sulfate (MnSO4·H2O) 0.25g.
[0031] 2. Preparation of bacterial suspen...
Embodiment 2
[0050] Example 2 Probiotic tolerance test
[0051] 1. Evaluation of acid resistance
[0052] The strains were inoculated in liquid MRS medium according to the inoculum volume of 3%, and cultured at 37°C for 18h. culture medium at 4°C at 4000rpm·min -1 Centrifuge for 10 minutes to obtain the bacteria sludge, wash twice with 0.85% normal saline, and then suspend the bacteria sludge with sterile normal saline to ensure that the activated bacterial strain is about 10 8 CFU mL -1 , and then inoculate the bacterial suspension in the MRS medium with a pH of 2.0 and 3.0 according to the inoculation amount of 3%, take samples at 0h, 2h, and 4h respectively, dilute 10 times with normal saline, and spread it on the MRS after diluting the appropriate concentration. In the solid medium, culture at 37°C for 48 hours, count the colonies, make three parallels for each concentration, calculate the average value, and calculate the number of viable bacteria and survival rate;
[0053] Result...
Embodiment 3
[0067] Embodiment 3 antioxidant activity test
[0068] 1.H 2 0 2 tolerance test
[0069] Inoculate the pure culture of Lactobacillus fermentum KP101 into MRS broth supplemented with different concentrations of hydrogen peroxide (0.5, 1.0 mmol / L); measure the absorbance at 600nm every 2h from 0h, and calculate the survival rate;
[0070] Results: The tolerance of the strain to hydrogen peroxide was concentration-dependent. The strain had strong tolerance to low concentration of hydrogen peroxide (0.5mM), and the survival rate of 37°C for 4 hours increased significantly. (1mM) the survival rate increased less, and the survival rate of the 8h strain was not less than 90%, indicating that Lactobacillus fermentum KP101 has strong tolerance to different concentrations of hydrogen peroxide ( Image 6 a).
[0071] 2. Clear DPPH free radicals
[0072] Centrifuge the bacterial liquid at 4000r / min for 10min to obtain the fermentation supernatant and bacterial cells respectively, was...
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