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A bifidobacterium strain that can be used for selenium-enriched culture and its active protein and application

A bifidobacterium and active protein technology, applied in the field of microorganisms, can solve the problems of affecting the intestinal colonization of bifidobacteria, reducing proton power, and destroying the osmotic pressure balance of bifidobacteria cells, so as to achieve good intestinal colonization ability, Effects of hepatoprotection, good gut protection and hepatoprotective function

Active Publication Date: 2020-06-19
JIANGSU DAYSEBIOTECH LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few bifidobacteria strains that can be used in food at home and abroad. This is because after bifidobacteria are orally ingested into the human body, various factors will affect their colonization in the human intestinal tract, for example, the low pH value of the stomach The high acidic environment in the environment and the digestive enzymes in it, the acidic environment will reduce the proton power and destroy the osmotic pressure balance inside and outside the bifidobacteria cells; the high concentration of bile salts and endogenous antimicrobial peptides in the small intestine and the intestinal bacteria The group's defense system to the body, glycocholate and bezoar bile salt have cell membrane toxicity, bifidobacteria with high activity of bile salt hydrolyzing enzyme can decompose bile salt into secondary bile salt, which can reduce its cell membrane toxicity; Bifidobacterium adhesion The ability to enter the intestinal epithelial cells and the ability to utilize nutrients that are not digested and metabolized by the host (some protein substances, prebiotics such as oligosaccharides, etc.) will affect the intestinal colonization of bifidobacteria

Method used

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  • A bifidobacterium strain that can be used for selenium-enriched culture and its active protein and application
  • A bifidobacterium strain that can be used for selenium-enriched culture and its active protein and application
  • A bifidobacterium strain that can be used for selenium-enriched culture and its active protein and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040]The acquisition of embodiment 1 bifidobacterium DD98 and strain isolation, screening

[0041] 1. Sample collection and expansion culture

[0042] Take 1g of the fresh mid-section feces of the sampled subjects, add 10ml of normal saline, mix well, let stand at room temperature for 5min, and collect the upper suspension. Take 1ml of the supernatant suspension, inoculate it into 50ml of RCM medium, and incubate anaerobically at 37°C for 24h to obtain a culture solution.

[0043] 2. Strain Isolation

[0044] The culture solution was taken and diluted 10 times with normal saline -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 times, to obtain a bacterial suspension, pipette 0.1ml of the bacterial suspension and spread it on the BBL medium, and culture it anaerobically at 37°C for 24 hours to obtain a single colony.

[0045] 3. Strain purification

[0046] Pick a single colony, streak and inoculate the colony culture on the BBL medium, and inoculate it anaerobically at 37°C for 24 ...

Embodiment 2

[0051] Example 2 Selenium-enriched domestication investigation of bifidobacterium DD98

[0052] Prepare a sodium selenite solution, filter it at 0.22um and add it to the culture medium, then distribute the culture medium to 96 deep-well plates and plates. Preliminary design of selenium-enriched acclimation gradient: 5, 10, 15, 20, 25, 50, 100, 200, 400, 600, 800 μg / ml.

[0053] Divide glycerol tubes of dominant strains of bifidobacteria into 96 deep-well plates containing 5 μg / ml sodium selenite medium, culture anaerobically for 24 hours, take 200 μl from each well of 96 deep-well plates to 96 microplate plates for OD600 For detection, the 3 wells with the highest OD value were diluted to 10 5 ~10 6 Double coated onto 5μg / ml sodium selenite plate. Put into anaerobic incubator and cultivate 18 hours after plate coating, after treating that single bacterium colony grows, pick the single bacterium colony (illustration that this bacterium colony grows well on the sodium selenit...

Embodiment 3

[0054] The bacteriological characteristic of embodiment 3 bifidobacteria DD98

[0055] 1. Colony Morphological Characteristics

[0056] Observing the isolated and purified DD98 with a scanning electron microscope, the bacteria are long rod-shaped or branched rod-shaped at the end, and the bacteria are generally about 10um in length, such as figure 1 shown.

[0057] The morphological characteristics of the colony: neat edges, soft texture, smooth surface, milky white protrusions, such as figure 2 shown.

[0058] Gram staining optical microscope observation: the staining result is purple, such as image 3 Shown, indicating Gram-positive bacilli.

[0059] 2. Biochemical characteristics

[0060] Using Bergey's Handbook as a reference, the characteristics of sugar fermentation and acid production of DD98 were investigated, and the results are as follows:

[0061]

[0062]

[0063] The results showed that: Bifidobacterium DD98 can utilize L-arabinose, D-galactose, D-fru...

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Abstract

The present invention discloses a novel Bifidobacterium DD98 having a 16SrDNA nucleotide sequence shown in SEQ ID No: 1. Further, the Bifidobacterium is preserved in the General Microbial Culture Collection Management Center of China, and a preservation number is CGMCC No. 16573. The Bifidobacterium DD98 of the invention can tolerate higher selenium pressure, can be used for selenium-enriched culture, and has good intestinal colonization ability, and the extracted water-soluble protein has intestinal protection function. The Bifidobacterium DD98 of the invention can alleviate liver damage caused by a high-fat diet and alcohol, and has the protective effect on the liver.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bifidobacterium and its active protein and its application. Further, the bifidobacteria can be prepared into selenium-enriched probiotics through selenium-enriched culture. Background technique [0002] Probiotics are a class of microorganisms that can have beneficial effects on the host and are closely related to human health. Probiotics generally colonize the intestinal tract and reproductive system of the human body, and play an important role in maintaining the micro-ecological balance in the human body. Existing research results have shown that the stability of human microbiota is very important for maintaining human health. Aging, hypertension, diabetes and intestinal diseases are all related to human microbiota. The imbalance of microbiota in the human body will directly or indirectly lead to the occurrence of these diseases. Bifidobacterium is a common physiolo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/04C12N1/36C07K14/195A61K35/745A61K38/16A61P1/00A61P29/00A61P1/12C12R1/01C12R1/16A23L33/135
Inventor 陈代杰邹振荣邵雷陆惠刚谭俊朱慧周艳纪瑞
Owner JIANGSU DAYSEBIOTECH LTD
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