Lactobacillus reuteri ZJUIDS09 for reducing blood pressure and application thereof
A Lactobacillus reuteri, blood pressure lowering technology, applied in the direction of Lactobacillus, application, bacteria used in food preparation, etc., can solve the problem that the blood pressure lowering effect of GMNL-263 live bacteria has not been proved, and achieve a high ACE inhibition rate. Effect
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Embodiment 1
[0034] Embodiment 1, screening and identification of Lactobacillus reuteri ZJUIDS09:
[0035] 1. Screening of Lactic Acid Bacteria
[0036] 1.1 Sample source
[0037] The bacterial strain used in the present invention is derived from milk fans made by farmers in Yunnan. A total of 15 samples were collected, which were immediately refrigerated and mailed to the laboratory for isolation and purification of the strains.
[0038] 1.2 Isolation and purification of strains
[0039] About 5 g of fresh samples were collected with sterile tubes and immediately sent to the laboratory for strain isolation. Take 1 g of the sample and put it into 9 mL of MRS liquid medium, vortex and mix it, and enrich and culture it at 37°C for 48 hours; then absorb 1 mL of the enriched solution in a super-clean bench, perform ten-fold gradient dilution with sterile normal saline, and select 10 -6 、10 -7 、10 -8 For three dilution gradients, 100 μL of bacterial solution was taken from each gradient ...
Embodiment 2
[0050] Example 2, Confirmation of the ability of Lactobacillus reuteri ZJUIDS09 to lower blood pressure
[0051] 1. ACE inhibition rate
[0052] Preparation of strain fermentation broth
[0053] Streak the strain ZJUID S09 stored in the glycerol tube to activate it 2-3 times on the MRS agar plate, then pick a single colony and expand it in the MRS liquid medium for 18 hours at 37°C, so that the concentration of the bacterial solution reaches about 10 8 About CFU / mL, as a culture medium. The culture solution was inoculated into sterilized reconstituted skim milk (11%, w / v) at an inoculum size of 2% (volume %), cultured at 37°C for 48h, taken out, and placed in a refrigerator at 4°C overnight. Take the fermented milk out of the refrigerator, adjust the pH to 4.6, centrifuge at 8000g for 10min to save the supernatant, adjust the pH to 8.2, centrifuge at 8000g for 10min, and use the collected supernatant as the fermentation supernatant of strain ZJUID S09.
[0054] The steriliz...
Embodiment 3
[0078] Embodiment 3, confirmation of the antioxidant capacity of Lactobacillus reuteri ZJUIDS09
[0079] The culture solution prepared in Example 2 was centrifuged at 8000r / min for 5min at 4°C to collect the bacteria, washed twice with phosphate buffer (pH 6.8), and suspended in the MRS liquid medium to make the live bacteria number corrected to about 10 8 CFU / mL, bacterial suspension;
[0080] The bacterial suspension and the fermentation supernatant prepared in Example 2 were used as samples. At the same time, Lactobacillus rhamnosus ATCC53103 was used as a positive control strain to measure the antioxidant activity.
[0081] 1. Total antioxidant capacity (FRAP method)
[0082] The determination method of total antioxidant capacity was slightly modified according to the method of Giuberti et al. Add 150 μL TPTZ working solution (0.3M acetic acid-sodium acetate buffer solution, 20mM ferric chloride solution, 10mM TPTZ buffer solution, mix in V:V:V=10:1:1, and prepare imme...
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