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Preparation method and application of triple probiotic fermentation compound with anti-inflammatory effect

A composition and functional technology, applied in the field of microbial fermentation, can solve the problem that probiotic fermentation products are difficult to achieve anti-inflammatory effects

Pending Publication Date: 2022-04-05
无锡弘焕微生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is: a single probiotic fermented product is difficult to achieve the desired anti-inflammatory effect

Method used

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  • Preparation method and application of triple probiotic fermentation compound with anti-inflammatory effect
  • Preparation method and application of triple probiotic fermentation compound with anti-inflammatory effect
  • Preparation method and application of triple probiotic fermentation compound with anti-inflammatory effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Screening of NO-resistant probiotics

[0034] Probiotic medium (g / L): peptone 10.0, yeast powder 5.0, glucose 20.0, beef extract 10.0, Tween-801mL, dipotassium hydrogen phosphate 2.0, sodium acetate 5.0, diammonium hydrogen citrate 2.0, magnesium sulfate heptahydrate 0.58, manganese sulfate 0.25, pH 6.5-6.8, solid contains 20g / L agar, sterilized at 121°C for 20min for later use.

[0035]NO resistance screening medium (g / L): peptone 10.0, yeast powder 5.0, glucose 20.0, beef extract 10.0, Tween-80 1mL, dipotassium hydrogen phosphate 2.0, sodium acetate 5.0, diammonium hydrogen citrate 2.0, heptahydrate Magnesium sulfate 0.58, manganese sulfate 0.25, sodium nitroprusside 2g / L, pH 6.5-6.8, solid contains 20g / L agar. Sterilize at 121°C for 20 minutes for later use.

[0036] Steps:

[0037] Step 1: Activation of strains

[0038] Put 24 strains of probiotics including Bifidobacterium plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus s...

Embodiment 2

[0049] Example 2: The test of the NO inhibition rate of the fermented products of the three probiotic strains screened in Example 1.

[0050] Steps:

[0051] step 1:

[0052] In the anaerobic glove box, pick a single colony of the three strains of NO-resistant probiotics obtained through screening and inoculate them into a shake flask containing 400 mL of sterile probiotic culture medium. Incubate at 37°C for 24h.

[0053] Step 2:

[0054] Preparation of probiotic fermentation filtrate: divide the cultured probiotic fermentation liquid into 50mL centrifuge tubes, centrifuge at 9000rpm for 10min, collect the supernatant, filter it with a 0.22μm filter membrane, and place it in a clean centrifuge tube to obtain the probiotic fermentation filtrate;

[0055] Preparation of probiotic lysate: Collect the sludge of each strain at the bottom of the centrifuge tube, resuspend the bacteria with an appropriate amount of sterile deionized water, centrifuge again at 9000rpm for 10 minu...

Embodiment 3

[0064] Example 3: Proportion test of fermented products of active probiotics with high NO inhibition rate.

[0065] The three strains of active probiotic lysates screened in Example 2 were compounded, and a combination with a synergistic NO inhibition rate was selected after mixing according to different mass ratios.

[0066] step 1:

[0067] Bifidobacterium H1 lysate (BH1L), Lactobacillus plantarum H2 lysate (PH2L), Lactobacillus acidophilus H4 lysate (AH4L) prepared in Example 2 were mixed according to the mass ratio shown in Table 3 , diluted 2 times with ultrapure water and used as a sample for later use.

[0068] Step 2:

[0069] NO inhibition rate test—take a 96-well plate, add 50 μL of 0.5 mM sodium nitroferricyanide dihydrate (SNP) solution, add 10 μL of the sample in step 1, and add Griess reagent after UV-A irradiation for 60 minutes A (1% sulfonamide dissolved in 2% phosphoric acid aqueous solution) 50 μL, Griess reagent B (0.1% N-(1-naphthyl) ethylenediamine hyd...

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Abstract

The invention discloses a preparation method and application of a triple probiotic fermentation compound with an anti-inflammatory effect, and belongs to the technical field of microbial fermentation. The triple probiotic fermentation compound disclosed by the invention is obtained by mixing lysate extracted after fermentation of three probiotic fermentation products with NO inhibition ratio, namely bifidobacterium H1, lactobacillus plantarum H2 and lactobacillus acidophilus H4, according to a ratio of 2: 1: 3. The traditional Chinese medicine composition disclosed by the invention has a very high anti-inflammatory effect, and can obviously reduce the expression of TNF-alpha in cells after being stimulated by LPS (Lipopolysaccharide) after being matched according to a synergistic effect, so that the anti-inflammatory effect is achieved.

Description

technical field [0001] The invention relates to a preparation method and application of a triple probiotic fermentation compound with anti-inflammatory effects, and belongs to the technical field of microbial fermentation. Background technique [0002] Probiotics are live microorganisms that, when ingested in adequate amounts, have a positive effect on the balance of the human microbiome. Probiotics Probiotics are the most common and easily accessible living microorganisms for humans. These active probiotics regulate the activity of reactive oxygen species (ROS) synthase and nitric oxide synthase (NOS) in the host through various mechanisms, such as the ability to chelate metal ions to act as antioxidants, making the oxidase system Vitality is down-regulated, thereby improving the body's defense ability to resist oxidative stress, reducing the expression of inflammatory factors, and achieving anti-inflammatory effects. [0003] SNP, also known as sodium nitroferricyanide, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K8/99A61Q19/00A61P29/00C12R1/25C12R1/23C12R1/01
CPCY02A50/30
Inventor 王斌罗西迟俐
Owner 无锡弘焕微生态科技有限公司
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