Lactobacillus paracasei LC86 and application thereof in prevention or treatment of decayed teeth and periodontitis
A technology of Lactobacillus and paracheese, applied in the field of microorganisms, can solve the problems of enhancing the drug resistance of oral pathogens, the risk of infection, and large side effects, and achieve the effect of improving the micro-ecological system, high tolerance, and strong self-aggregation ability
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Embodiment 1
[0026] Example one experiment of Lactobacillus paracasei LC86 inhibiting oral pathogenic bacteria
[0027] Streptococcus mutans ATCC 25175 used in this example was purchased from China General Microorganism Culture Collection and Management Center. Porphyromonas gingivalis ATCC BAA-308 was purchased from the Guangdong Provincial Microbial Culture Collection Center. The strains of Lactobacillus paracasei were obtained from the Weikang Probiotics Resource Bank.
[0028] MRS medium ingredients: yeast powder 5.0g / L, beef extract 10.0g / L, peptone 10.0g / L, glucose 20.0g / L, anhydrous sodium acetate 2.0g / L, diammonium hydrogen citrate 2.0g / L, Dipotassium hydrogen phosphate trihydrate 2.6g / L, manganese sulfate monohydrate 0.25g / L, magnesium sulfate heptahydrate 0.5g / L and Tween-80 1mL / L, pH 6.2~6.4.
[0029] BHI medium ingredients: bovine brain infusion powder 4.0g / L, beef heart infusion powder 4.0g / L, peptone 5.0g / L, casein peptone 16.0g / L, sodium chloride 5.0g / L, glucose 2.0g / L, D...
Embodiment 2
[0035] Example 2 Molecular biological identification of Lactobacillus paracasei LC86
[0036] The Lactobacillus paracasei strain was extracted by genomic DNA extraction kit, and the 16S rDNA sequence was amplified by using the upstream primer 27F (AGTTTGATCMTGGCTCAG) and the downstream primer 1492R (GGTTACCTTGTTACGACTT) to obtain the PCR product. and sequence the PCR products. The PCR reaction system: 20 μL of 10×Buffer, 4 μL of primer dNTP, 1 μL of upstream and downstream primers, 2 μL of DNA template, 0.5 μL of Taq enzyme, ddH 2 O 34.5 μL. PCR reaction conditions: 95°C for 10 min; 94°C for 30s, 56°C for 30s, 72°C for 2min, 35 cycles; 72°C for 10min. The PCR products were detected by gel electrophoresis and sent to Wuhan Jinkarui Bioengineering Co., Ltd. for sequencing. The identified gene sequences were submitted to the NCBI database (www.ncbi.nlm.nih.gov) for BLAST analysis and alignment. According to the identification results of molecular biology, the strain was ident...
Embodiment 3
[0037] Embodiment three Lactobacillus paracasei LC86 tolerance detection to lysozyme
[0038] Lactobacillus paracasei LC86 glycerol tube strain was inoculated into MRS liquid medium at 2% of the volume of MRS liquid medium, and after culturing at 37°C for 24 hours, the concentration of bacterial suspension was adjusted to 10 8 CFU / mL. Take 200uL of the above Lactobacillus paracasei LC86 bacterial solution and spread it on the MRS plate. After absorption, gently place the sterilized Oxford cup on the MRS plate coated with the bacterial solution. Then add 100uL of different concentrations of lysozyme solutions (0.1mg / ml, 0.3mg / ml, 0.5mg / ml, 0.7mg / ml, 1mg / ml, 2mg / ml) in the Oxford cup to make the solution without lysozyme. Bacterial water was used as a control, and each group was set up with 3 parallels. The above plates were stably placed in an anaerobic incubator at 37°C for 16-20 hours, and the size of the inhibition zone of Lactobacillus paracasei to lysozyme was detected. ...
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