Lactobacillus casei and composition thereof
A technology of Lactobacillus casei and a composition, which is applied in the field of microorganism application, can solve the problems such as the attenuation of the number of viable bacteria, and achieve the effects of high number of viable bacteria, good compounding effect, and good synergistic growth effect.
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Embodiment 1
[0025] The acquisition of embodiment 1 bacterial strain
[0026] Lactobacillus casei, isolated from healthy human intestines in Jianggan District, Hangzhou City, screened out strains suitable for human body, the preservation number of Lactobacillus casei is CGMCC No.4520, on December 29, 2010, preserved in China Microbiology in Beijing Culture Collection Management Committee General Microbiology Center.
[0027] Lactobacillus casei was collected from 20 healthy children aged 2-6 years old in Jianggan District, Hangzhou City, 10 males and 10 males, all had no history of gastrointestinal diseases, and all of them took any antibacterial drugs within two weeks before sampling. When sampling, use a sterile glass rod to pick up 4-10g of naturally discharged fresh feces, place them in a sterile plate, quickly put them into an anaerobic tank, and bring them back to the laboratory for separation.
Embodiment 2
[0028] Identification and cultivation of embodiment 2 bacterial strains
[0029] 2.1 Use the dilution coating method to separate and purify the collected strain samples. After purification and culture, observe the colony morphology, take fresh culture, inoculate sugar fermentation test tube, and identify the biochemical reaction characteristics of the strain. At the same time, the fresh culture of the strain to be tested was taken, and the API biochemical identification of the strain to be tested was carried out by using the API 50CHL biochemical identification strip of the French Mérieux company, referring to the biochemical identification database of the Mérieux company. At the same time, the 16SrRNA of the isolated and purified strain will be extracted and sequenced, and the sequencing results will be compared with the existing sequences in NCBI's GeneBank. The isolated and purified strains were stored at 2-8°C.
[0030] After identification, the colonies of the strains t...
Embodiment 3
[0048] 3.1 Bile salt tolerance test
[0049] The composition of the improved MRS nutrient solution used for cultivating bacterial strains is:
[0050] The isolated and purified bacterial strains were cultured strictly anaerobically at 37° C. for 15 h with sterilized modified MRS medium. The culture solution of the activated strain was inserted into the sterile modified MRS liquid medium containing different bile salt concentrations with 10% inoculum (the bile salt concentrations were 1%, 2%, 3%, 4%, 5%, 6%, 7%, %), while the improved MRS culture medium without bile salts was used as a control. After anaerobic culture at 37°C for 0, 6, 12, 18, and 24 hours, samples were taken to determine the number of viable bacteria.
[0051] 3.2 Acid tolerance test
[0052]The isolated strains were cultured strictly anaerobically at 37° C. for 15 h with sterile modified MRS medium. The culture solution of the activated strain was inserted into the sterile modified MRS culture solution wi...
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