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 prob

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

Example Embodiment

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

[0034] Probiotic culture 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 containing 20g / L agar, sterilized at 121°C for 20min for use.

[0035]NO tolerance 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 containing 20g / L agar. Sterilize at 121°C for 20 min.

[0036] Steps:

[0037] Step 1: Strain activation

[0038] A total of 24 strains of probiotics including Bifidobacterium, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacil...

Example Embodiment

[0049] Example 2: Inhibition rate test on NO of three probiotic fermented products screened in Example 1.

[0050] Steps:

[0051] step 1:

[0052] The 3 strains of NO-resistant probiotics obtained by screening were picked in an anaerobic glove box, and a single colony was picked and inoculated into a shake flask containing 400 mL of sterile probiotic culture medium. The size of the shake flask was 500 mL and placed in an anaerobic incubator. medium, incubated at 37°C for 24h.

[0053] Step 2:

[0054] Preparation of probiotic fermentation filtrate: The cultured probiotic fermentation broth was divided into 50mL centrifuge tubes, centrifuged at 9000rpm for 10min, the supernatant was collected, filtered with a 0.22μm filter membrane, and placed in a clean centrifuge tube to obtain probiotic fermentation filtrate;

[0055] Preparation of probiotic lysate: collect the bacterial sludge of each strain at the bottom of the centrifuge tube, resuspend the bacteria with an appropri...

Example Embodiment

[0064] Example 3: Proportioning test of active probiotic fermentation products with high NO inhibition rate.

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

[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 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 min A (1% sulfanilamide dissolved in 2% phosphoric acid aqueous solution) 50 μL, Griess reagent B (0.1% N-(1-naphthyl)ethylenedi...

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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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