Lactobacillus paracasei capable of relieving alcoholic intestinal injury and application of lactobacillus paracasei capable of relieving alcoholic intestinal injury
A technology of para-cheese and lactobacillus, applied in the field of microorganisms, can solve the problems of slow effect and low safety, and achieve the effect of improving pathological damage and high safety
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[0062] The preparation method of the lactobacillus suspension involved in the following examples is as follows:
[0063] Streak the Lactobacillus on the MRS solid medium and culture it at 37°C for 48 hours to obtain a single colony; pick a single colony and inoculate it in the MRS liquid medium, culture it at 37°C for 18 hours for activation, and activate two generations continuously to obtain Activation solution; the activation solution was inoculated in MRS liquid medium at an inoculum size of 2% (v / v), and cultured at 37°C for 18 hours to obtain a bacterial solution; the bacterial solution was centrifuged at 4°C and 6000g for 10 minutes, The Lactobacillus cells were obtained; the Lactobacillus cells were washed 3 times with normal saline and then resuspended in a skimmed milk powder solution with a concentration of 120g / L to a bacterial concentration of 1×10 9 CFU / mL to obtain a bacterial suspension, and store the bacterial suspension at -80°C until use.
Embodiment 1
[0064] Embodiment 1: Screening and identification of Lactobacillus paracasei CCFM1120
[0065] 1. Screening
[0066] Take fresh feces from healthy people in Kunshan City, Jiangsu Province as the sample. After the sample is pretreated, it is stored in a -80°C refrigerator in about 30% glycerol. Perform gradient dilution with 0.9% normal saline, select the appropriate gradient dilution and spread it on the MRS solid medium, incubate at 37°C for 48 hours, pick typical colonies to streak and purify on the MRS plate, pick a single colony and transfer it to liquid MRS The liquid medium was enriched and preserved in 30% glycerol to obtain strain CCFM1120 (the original number of the strain was RS85).
[0067] 2. Identification
[0068] The genome of CCFM1120 was extracted, and the 16S rDNA of CCFM1120 was amplified and sequenced (completed by Shanghai Sangon Bioengineering Co., Ltd.). After sequencing analysis, the 16S rDNA sequence of the bacterial strain was shown in SEQ ID NO.1. ...
Embodiment 2
[0069] Embodiment 2: the cultivation of Lactobacillus paracasei CCFM1120
[0070] After inserting Lactobacillus paracasei CCFM1120 into the MRS solid medium and culturing at 37°C for 48 hours, the colonies were observed and observed under a microscope. It was found that the colonies were milky white semicircular protrusions with smooth and moist surfaces. Neat edges.
[0071] Lactobacillus paracasei CCFM1120 was inserted into MRS liquid medium and cultured at 37°C for 48 hours. During the cultivation process, the pH of the culture solution was measured by a pH meter at intervals, and it was found that Lactobacillus paracasei CCFM1120 produced acid during the cultivation process.
[0072] Insert Lactobacillus paracasei CCFM1120 into MRS liquid medium and culture them at 10-50°C for 48 hours. During the cultivation process, measure the OD of the culture solution with a microplate reader at intervals 600 , it was found that Lactobacillus paracasei CCFM1120 grew best at 30-37°C, ...
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