Lactobacillus rhamnosus capable of relieving chronic alcohol liver injury and application thereof
A kind of Lactobacillus rhamnosus, the technology of use, applied in the field of microbiology
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Embodiment 1
[0083] Embodiment 1: 16S rDNA sequence identification of Lactobacillus rhamnosus CCFM1107 of the present invention
[0084] Lactobacillus rhamnosus CCFM1107 was inoculated in MRS liquid medium, and cultured at 37°C for 18 hours. Take 1 mL of the culture liquid and use the bacterial genome DNA extraction kit to operate according to its instructions. Genomic DNA was used as a template, and bacterial identification universal primers published in literature (Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes. Applied and Environmental Microbiology, 2008, 74(8): 2461-2470) were used as primers, and 50 μL The reaction system is amplified by PCR, and then the amplified product is purified, recovered and sequenced.
[0085] The sequencing of the target gene PCR amplification product was completed by Shanghai Bioengineering Technology Service Co., Ltd. The sequencing results of the 16S rDNA of Lactobacillus rhamnosus CCFM1107 were compared us...
Embodiment 2
[0086] Example 2: Determination of microbiological properties of Lactobacillus rhamnosus CCFM1107
[0087] Lactobacillus rhamnosus CCFM1107 was inserted into the MRS liquid medium according to the inoculation amount of 5% based on the volume of the MRS liquid medium. At these time points of 18h, 20h, 22h and 24h, the pH value of the culture solution was measured with a pH meter, and the OD at 600nm was measured with a spectrophotometer 600 value. The pH meter that the present invention uses is the 320-S pH meter that Mettler-Toledo Instrument (Shanghai) Co., Ltd. sells, and the spectrophotometer that uses is the UV-2100 ultraviolet-visible spectrometer that Unico (Shanghai) Instrument Co., Ltd. sells Photometer.
[0088] by OD 600 Value and pH value are plotted against culture time, and the growth curve of Lactobacillus rhamnosus CCFM1107 in MRS medium can be obtained, and the results are as follows image 3 shown. In the MRS medium, the lag phase of Lactobacillus rhamnos...
Embodiment 3
[0089] Example 3, the antioxidant capacity of Lactobacillus rhamnosus CCFM1107
[0090] First, whole cells and cell-free extracts of Lactobacillus rhamnosus CCFM1107 were prepared.
[0091] Inoculate Lactobacillus rhamnosus CCFM1107 of the present invention into MRS liquid medium after activated culture, culture at 37°C for 24h, centrifuge at 6000r / min and 4°C for 10min to obtain culture supernatant and cell precipitation, the cell pellet was washed twice with sterile normal saline, resuspended in sterile normal saline, and the cell concentration was adjusted to about 10 9 cfu / mL. The obtained bacterial cell suspension was divided into two groups, one group was the complete cell group (IC), and the other group was used for the preparation of the cell-free extract (CFE). The cell suspension was ultrasonically crushed with an ultrasonic breaker (Sonics & Materials Company, VCX500 type) at 200W and a temperature of 4°C. Each treatment was carried out for 5s, and the interval be...
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