Lactobacillus fermentum and application thereof
A technology of fermenting Lactobacillus and live bacteria, which is applied in the field of microorganisms and can solve the problems of reducing the water content of the cuticle
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Embodiment 1
[0044] Example 1 Separation of ProfMIC-205
[0045] Sampling in soy juice. After the samples were properly treated, they were shaken and mixed in normal saline. The supernatant was taken and streaked on the MRS solid plate. After incubation at 37°C for 24-48 hours, white colonies were picked and repeatedly inoculated and screened until a uniform single colony was obtained, which was named ProfMIC. -205.
[0046] Gram stain microscopy: The strain ProfMIC-205 is Gram stain positive and is rod-shaped under the microscope; it grows on the MRS plate and can form small white, round and opaque small colonies with a smooth surface and neat edges; cultured in MRS liquid In the base, the growth is uniform and turbid, and the bacteria are white precipitated after being left for a long time.
Embodiment 2
[0047] Example 2 Nucleic acid identification of ProfMIC-205
[0048] 1. 16s rRNA gene sequence analysis:
[0049] A single colony was picked and placed in MRS liquid medium, and after culturing overnight at 37°C, the cells were collected by centrifugation, and the operation was performed according to the steps of the DNA extraction kit. Primers were bacterial universal primers 27F, 1492R, PCR amplification system was 50 μL system, pre-denaturation at 95°C for 5min; 94°C for 15s, 57°C for 15s, 72°C for 40s, 35 cycles; extension at 72°C for 10min.
[0050] 2. Results
[0051] The PCR product sequencing results were compared with the standard sequences published in GenBank (BLASTN), and it was concluded that the ProfMIC-205 strain was Lactobacillus fermentum.
Embodiment 3
[0052] Example 3 ProfMIC-205 promotes SDS-induced damage and repair of HaCaT cells
[0053] 1. Preparation of ProfMIC-205 bacterial solution:
[0054] The activated Lactobacillus fermentum PROFMIC-205 strain was cultured in MRS liquid medium in a 37°C incubator for 16-18 hours, and the OD was detected and adjusted. 600 = 2.0, 121°C, 30min inactivation, centrifugation to take the precipitate as inactivated bacteria, and the inactivated fermentation broth was filtered through a 0.22 μm filter to obtain the inactivated supernatant.
[0055] 2. Promoting HaCaT cell manipulation and repair experiment
[0056] Seed HaCaT cells (5×10 4 cells / well) to a 96-well plate and cultured overnight until cells adhered. Prepare 50 μg / ml SDS, add 100 μl to each well, incubate for 8 h, add 5% supernatant and 10% inactivated bacteria respectively, and incubate for 24 h. Add 10 μl of CCK-8 solution, incubate for 4 h, and detect the absorbance A at 450 nm.
[0057] Survival rate=experimental gr...
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