Lactobacillus plantarum with oxidation resistance and application thereof
A technology of Lactobacillus plantarum and anti-oxidation ability, applied in the direction of Lactobacillus, medical preparations containing active ingredients, applications, etc., can solve the problems of biological macromolecule damage, and achieve the effect of improving activity, reducing content, and good probiotic properties
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Embodiment 1
[0045] Example 1: Isolation and identification of lactic acid bacteria from traditional fermented yak yoghurt and silage on the Qinghai-Tibet Plateau 1. Sample source
[0046] It comes from traditional fermented yak dairy products and silage in Tianzhu Tibetan Autonomous County, Gansu Province, my country.
[0047] 2. Isolation of lactic acid bacteria from traditional yak dairy products and silage
[0048] Take 1mL of fermented yak milk product and dilute it with sterilized saline at a ratio of 1:10, which is 10 -1 dilution. Then draw 1mL of the above diluent, and then dilute it 10 times, which is 10 -2 dilution, and so on. Pick 10 -5 、10 -6 、10 -7 For three dilutions, draw 100 μL of the dilutions and spread them evenly on the MRS, M17, KFS and SL (adjust pH 5.4) plate culture medium respectively. ℃ and 37 ℃ anaerobic constant temperature culture for 48 hours, pick the characteristic colonies, and further purify to obtain pure isolated lactic acid bacteria strains.
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Embodiment 2
[0056] Embodiment 2: Screening of high antioxidant capacity lactic acid bacteria
[0057] 1. Preparation of complete cell and cell-free extracts and fermentation supernatant of lactic acid bacteria
[0058] Preparation of cell bacterial liquid: the bacterial liquid stored at -80°C in the laboratory was inserted into the sterilized MRS medium and cultivated overnight at 37°C, and was continuously passaged for 3 times. The culture solution was collected by centrifugation at 12,000 g, washed with PBS three times, resuspended in PBS, and the number of bacteria was adjusted.
[0059] Preparation of cell-free extract: activate the bacterial solution twice overnight at 37°C, centrifuge at 8000g, 10min, 4°C, collect the bacterial cells, wash with PBS for 3 times, resuspend in PBS, adjust the number of bacteria to 1×10 9 CFU / mL or 1×10 10 CFU / mL. Sonicate the cells in an ice bath, 5s, 5s, 30min, 360W. 8000g, 10min, 4°C, collect the supernatant as a cell-free extract.
[0060] Prep...
Embodiment 3
[0084] Example 3: In vitro studies on probiotic properties
[0085] 1. The digestive tract simulation test and hydrophobicity determination of the invented strain
[0086] (1) Hydrophobic ability
[0087] According to the method of Mansona et al. (1992), the activated lactic acid bacteria were inoculated into liquid MRS medium, cultivated at 37°C for 16-18h, centrifuged at 5000rpm / min, and washed with 0.1mol KNO 3 (pH6.2) After washing twice, use the solution to suspend the bacterial liquid, and measure its initial absorbance A at 600nm 0 . Then draw 3ml of bacterial suspension, add 1ml of xylene, pre-incubate at room temperature for 10min, vortex quickly mix for 2min, let stand at room temperature for 15min to separate layers, absorb the lower aqueous phase, use the buffer as a blank control, measure the final absorbance A at 600nm and Make a note.
[0088] Hydrophobic capacity (%) = (A 0 -A) / A 0 ×100%
[0089] In the formula: A 0 and A are the absorbance values mea...
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