Lactobacillus gasseri and application thereof to preparation of fermented milk
A technology of Lactobacillus gasseri and fermented milk, which is applied in the field of fermented dairy products to achieve the effects of high number of viable bacteria, inhibition of inflammatory factors and strong antioxidant capacity
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Embodiment 1
[0035] The isolation and identification of embodiment 1 bacterial strain
[0036] The isolation method of Lactobacillus gasseri, the main steps are as follows:
[0037] The intestinal feces collected from 1-month-old infants in Guangzhou were used as samples. After the samples were serially diluted, three appropriate serial dilutions were selected, spread on the MRS medium, and the plate was placed upside down in an anaerobic incubator at 37°C for 48 hours. . According to the characteristics of colony size and shape, the strains similar to Lactobacillus were selected for microscopic examination and isolated and purified.
[0038] The DNA of the strain was extracted, the 16S rDNA sequence was sequenced, and compared with the BLAST software on the NCBI database, it was found that one strain had a homology of more than 99% with the 16S rDNA sequence of Lactobacillus gasseri. The 16S rDNA sequence of Lactobacillus gasseri LGZ 1029 of the present invention is shown in SEQ ID NO:1...
Embodiment 2
[0041] Embodiment 2 Lactobacillus gasseri LGZ 1029 acute toxicological test
[0042] (1) Experimental group reagent: the test substance is an oral liquid containing Lactobacillus gasseri LGZ 1029, the concentration is 1×10 8 CFU / mL, pale yellow liquid.
[0043] (2) Animals and their feeding environment: Take 20 SPF grade KM mice, weighing 18-22 g, provided by Jinan Pengyue Experimental Animal Breeding Co., Ltd., experimental animal production license number: SCXK (Lu) 2014-0007, Quality certificate number: No.37009200020118. The feed was provided by the Guangdong Medical Experimental Animal Center. The animals are used after passing the quarantine, the use license number: SCXK (Guangdong) 2018-0137, the temperature of the animal room is set at 20-26°C, and the relative humidity is kept at 40-70%.
[0044] (3) This experiment adopts the limit method, and only one dose group of 10.0 g / kg body weight is set, that is, 20 mice that pass the inspection and quarantine are taken, h...
Embodiment 3
[0048] The tolerance determination of embodiment 3 Lactobacillus gasseri LGZ 1029
[0049] 1. Determination of tolerance to simulated gastrointestinal fluid
[0050] Tolerance evaluation of Lactobacillus gasseri LGZ 1029 in artificial gastric juice: Accurately measure 20mL of 1mol / L hydrochloric acid, adjust the pH to 2.0, 2.5, 3.0, 4.0 by adding distilled water and 1mol / L NaOH solution, and then Add pepsin at a concentration of 5 g / L, fully dissolve, and then filter and sterilize with a microporous membrane with a pore size of 0.22 μm to prepare artificial gastric juice for future use. Take Lactobacillus gasseri activated in MRS medium and inoculate it in the above-mentioned artificial gastric juice according to the inoculum amount of 5% (v / v), then cultivate it at 37°C for 3 hours, take samples every 1 hour and test the live bacteria Count, set a blank control group, and repeat the parallel experiment 3 times. The ratio of the logarithmic value of the number of viable bact...
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