Lactobacillus plantarum and its uses for eliminating body fat, reducing hepatomegaly and anti-inflammation
A technology for the use of Lactobacillus plantarum, which is applied in the field of Lactobacillus plantarum strains to prevent and/or treat obesity-related diseases, and can solve problems such as inability to meet the needs of relief
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Embodiment 1
[0091] Embodiment one: collection and separation and purification of strains
[0092] Visit farmers in Yilan, Hualien, Taitung and other places engaged in farming, and observe senior farmers over 75 years old who are still self-sufficient and flexible in farming. It is inferred that long-term intake of foods containing lactic acid bacteria, that is, long-term exposure to strains with unique beneficial effects, will help elderly farmers maintain good physiological functions. Therefore, samples that may contain lactic acid bacteria are collected from various places, and the samples are stored in sealed sterile bags, refrigerated in a mobile refrigerator, and brought back to the laboratory to isolate and purify the probiotic strains contained in the samples. First, prepare a medium containing different nutrient sources to screen out different types of lactic acid bacteria strains in the sample. Examples of the components and types of the medium are: MRS agar, PCA agar, M17 agar, ...
Embodiment 2
[0094] Embodiment two: the genetic analysis and identification of strain
[0095] Firstly, the DNA of GKM3 was collected for sequencing, and the strain was identified by gene comparison. Streak activation of GKM3, use the inoculation loop to pick up a single colony and inoculate it into the medium for amplification. Collect fresh bacteria the next day and extract strain gDNA by CTAB method, and confirm the quality of gDNA by 0.6% agar gel electrophoresis. After concentration, the third-generation sequencing technology platform PacBio RSII was used to analyze the whole genome of the strain. According to the analysis results of the whole genome, the whole genome of GKM3 is 3,076,381 bp in length and contains a 49,415 bp plasmid (SEQ ID NO: 1). The whole genome sequence of GKM3 was compared with the whole genome database, and the pheS (SEQ ID NO: 2) and recN (SEQ ID NO: 3) gene sequences of GKM3 were obtained. Next, the sequence of the pheS gene (SEQ ID NO: 2) of GKM3 was comp...
Embodiment 3
[0102] Embodiment three: strain culture
[0103] The effects of different culture conditions on the production of GKM3 were studied. The carbon source and nitrogen source in the fermentation medium for cultivating GKM3 were tested with a one-factor test method at a time.
[0104] First, starting from the demand for carbon sources, compare the number of colonies of GKM3 and the IFN-γ of peripheral leukocytes and lymphocytes under the conditions of 8 carbon sources: glucose, sucrose, lactose, fructose, mannose, sorbitol, glycerol and molasses Inducible ability to select a carbon source with high yield and high physiological activity of GKM3. In the MRS medium, 3 wt% of the above 8 carbon sources were added relative to the total weight of the medium, and the culture temperature was set at about 37° C. and the rotation speed was about 20 rpm. The preferred 4 carbon sources and the number of colonies formed by viable bacteria in the experimental results are shown in Table 2 below...
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