Akkermansia muciniphila 139 strain and purpose thereof

A technology of strains and uses, applied in the field of microorganisms, can solve problems such as inflammation and immunity in mice, and achieve the effects of promoting Treg cell differentiation, regulating permeability and improving health.

Active Publication Date: 2020-06-23
上海究本科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on Akkermansia muciniphila ATCC BAA-835 mainly focuses on its effects on obesity and diabetes in mice and its mechanism, and less on its effects on inflammation and immunity in mice
At the same time, Akkermansia muciniphila ATCC BAA-835 is a human strain that does not exist in the original intestine of mice, so its role in mice may be related to Akkermansia muciniphila colonized in the mouse intestine itself. strains vary

Method used

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  • Akkermansia muciniphila 139 strain and purpose thereof
  • Akkermansia muciniphila 139 strain and purpose thereof
  • Akkermansia muciniphila 139 strain and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Isolation and identification of Akkermansia muciniphila 139 strain

[0031] Fresh feces of male C57 / BL6 mice of specific pathogen free (SPF) grade were selected and diluted in a gradient from 10-3 to 10-13, and then the bacterial solutions of different dilutions were spread on the surface of the culture medium respectively. To cultivate. A plate with more and clear colonies was selected, and all colonies were identified by PCR using Akkermansia muciniphila species-specific primers. The positive colonies were streaked and purified three times, and finally identified by Sanger sequencing using denaturing gradient gel electrophoresis (DGGE), ERIC fingerprinting and full-length 16S rRNA gene.

[0032] Wherein, the sequence of Akkermansia muciniphila species-specific primer is as follows:

[0033] AM1: 5'-CAGCACGTGAAGGTGGGGAC-3' (SEQ ID NO: 1)

[0034]AM2: 5'-CCTTGCGGTTGGCTTCAGAT-3' (SEQ ID NO: 2)

[0035] The 25µL PCR reaction system is: 2.5µL of 10× buffer, 2...

Embodiment 2

[0043] Morphology and growth curve of embodiment 2 Akkermansia muciniphila 139 bacterial strain

[0044] The morphology of the Akkermansia muciniphila 139 bacterial strain obtained by the separation of the above-mentioned embodiment 1 is observed by a scanning electron microscope, and it is observed that the bacterial cell size of the bacterial strain is about 0.9 × 0.5 μm, and the surface is smooth, oval, and without flagella, such as figure 2 shown.

[0045] The growth curves of the Akkermansia muciniphila 139 strain were measured in Mucin medium and synthetic medium respectively, the fermentation system was 200ml, and 3 repetitions were set at each time point, and the measurement results were expressed as mean ± standard error. The results of the growth curve measurement were as follows: image 3 shown by image 3 It can be seen that the strain grows faster in the Mucin medium, reaching the plateau in about 23 hours, and the growth amount in the plateau is about 1.58×109...

Embodiment 3

[0046] Example 3 Akkermansia muciniphila 139 strain in vitro fermentation

[0047] The pH value of the fermentation supernatant after the above-mentioned Akkermansia muciniphila 139 strain reached plateau in Mucin medium and synthetic medium was measured respectively. The results showed that in these two mediums, the fermentation of Akkermansia muciniphila 139 made the fermentation supernatant A significant drop in pH occurred (see Table 2).

[0048] Table 2 Changes in pH value of Akkermansia muciniphila 139 strain reaching plateau

[0049]

[0050] Note: The fermentation system is 200ml, and each group has 3 repetitions, and the measurement results are expressed as mean ± standard error.

[0051] In order to further study the reasons for the drop in pH value, gas chromatography (Agilent, USA) was used to measure the content of short-chain fatty acids in the fermentation supernatant of the Akkermansiamuciniphila 139 strain when it was fermented to the plateau stage in Muci...

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Abstract

The invention discloses a new Akkermansia muciniphila 139 strain. The strain is collected in the China General Microbiological Culture Collection Center with a collection number of CGMCC No. 16758, and a collection date is November 21st, 2018. The invention also discloses a purpose of the strain. Through separation and authentication, a new strain, i.e., the Akkermansia muciniphila 139, of Akkermansia muciniphila is obtained, the strain can carry out fermentation in vitro to generate a great quantity of short chain fatty acids, and the intestinal health of a host can be maintained. The fermentation in vitro of supernatant can induce the differentiation of mice lymphocyte regulative T cells, and the immunologic function of the host is enhanced. In addition, the bacteria strain also can alleviate enteritis, especially DSS (dextran sulfate sodium)-induced chronic enteritis, and also has a probiotic function.

Description

technical field [0001] The present invention relates to the field of microbes, in particular to Akkermansia muciniphila, and more specifically to the murine Akkermansia muciniphila 139 strain with probiotic effect and the use of the strain. Background technique [0002] Akkermansia muciniphila is a kind of intestinal microorganism that can colonize the intestinal mucosa with intestinal mucosal protein as the sole carbon source. Studies have shown that Akkermansia can play a beneficial role in various disease models (such as obesity, diabetes, alcoholic liver disease, and cardiovascular disease, etc.). At the same time, one of its extracellular vesicle extracts can alleviate DSS-induced acute colitis in mice, and by increasing intestinal permeability, it can enhance the intestinal barrier function of the host. Therefore, Akkermansia is considered as a strain with potential to be a probiotic. [0003] At present, there are very few Akkermansia that have been isolated and ide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/741A61P1/00C12R1/01
CPCA61P1/00A61K2035/115C12R2001/01C12N1/205
Inventor 张晨虹翟芮
Owner 上海究本科技有限公司
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