Lactobacillus casei with antihypertensive effect and application thereof
A Lactobacillus casei and blood pressure-lowering technology, applied in the field of functional bacteria, can solve the problems of low ACE inhibition rate, etc., and achieve the effects of high ACE inhibitory activity, strong inhibitory effect, and good acid resistance and bile salt resistance.
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Embodiment 1
[0030] The Lactobacillus casei SLK30 provided by the present invention is derived from a fermented feed sample. The strain is sequenced and analyzed, and the sequence obtained by sequencing is compared in the EzBioCloud Database. The result shows that the strain is Lactobacillus casei, and it is named Cheese Lactobacillus casei SLK30. Its microbial morphological characteristics are: round, milky white and translucent, smooth surface, convex, and neat edge. Microscopic observation appears in pairs or chains, belonging to Gram-positive strains. Bacterial form such as figure 1 shown.
[0031] Hemolytic capacity assay
[0032] Lactobacillus casei SLK30 was inoculated into sterilized MRS broth medium at 2% inoculation amount, cultured at 37° C. for 24 h, activated to 2 generations, and the strain vitality was restored to normal, ready for use.
[0033] Lactobacillus casei SLK30 was inoculated into a blood agar plate containing 5% sterile defibrillated sheep blood with a one-tim...
Embodiment 2
[0035] GABA-producing ability assay
[0036] Add 1% L-sodium glutamate to the MRS broth medium, sterilize by autoclaving at 121°C for 15 min, and inoculate the Lactobacillus casei SLK30 activated for 2 generations in Example 1 at an inoculum of 3% in the MRS medium. , after standing at 37°C for 24h, centrifugation (5200r / min, 20min), collecting the fermentation supernatant and storing it in a -40°C refrigerator for later use.
[0037] GABA standard dilution sample concentration: 0.1g / L, 0.2g / L, 0.4g / L, 0.6g / L, 0.8g / L; take 200μL of GABA standard dilution samples respectively, add 600μL of OPA derivatization reagent, and add 800μL after mixing KH 2 PO 4 Buffer, vortex mixed, filter through organic membrane, and wait for injection.
[0038] Chromatographic column: Agilent C-18 column; mobile phase A (15mmol / L CH 3 COOH-CH 3 COONa buffer): mobile phase B (methanol)=55:45; 0.8 mL / min; 30° C.; 334 nm; automatic injection; the injection volume was 20 μL.
[0039] Obtain the ch...
Embodiment 3
[0042] Ability to inhibit ACE rate
[0043] The 2-generation activated Lactobacillus casei SLK30 in Example 1 was inoculated into sterilized 12% skimmed milk powder according to the inoculum amount of 3%, and fermented at 42° C. for 34 hours to obtain fermented milk. The fermented milk was centrifuged at 4500 g for 10 min. The supernatant was taken, and the pH was adjusted to 7.5 with 1.0 mol / L NaOH, and the inhibitory activity of Lactobacillus casei SLK30 on ACE was determined by ACE kit. The assay method of ACE inhibitory activity was determined according to the method in the instructions of the ACE kit. The results showed that the inhibitory rate of Lactobacillus casei SLK30 on ACE was 89.99%, and the inhibitory activity on ACE was high.
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