Strain for preventing and treating metabolic diseases and application for strain

A technology for metabolic diseases and bacterial strains, which is applied in the field of microorganisms in the field of biotechnology, can solve the problems of Akkermansia strains' fermentation performance, storage stability, acid resistance and bile salt resistance, and the effect of treating metabolic diseases is not particularly ideal, to achieve Effects of lowering blood sugar, reducing visceral fat content, and delaying weight gain

Active Publication Date: 2020-04-07
SHANGHAI SINE PHARMA LAB +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the existing Akkermansia strain ATCC BAA835 that exists in the prior art in terms of fermentation performance, storage stability, acid resistance and bile salt resistance, and the effect on the treatment of metabolic diseases is not special. Ideal problem, while providing a bacterial strain and its application for the prevention and treatment of metabolic diseases

Method used

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  • Strain for preventing and treating metabolic diseases and application for strain
  • Strain for preventing and treating metabolic diseases and application for strain
  • Strain for preventing and treating metabolic diseases and application for strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the screening of bacteria

[0030] 1. Medium

[0031] Separation medium: 0.4g KH 2 PO 4 ; 0.53g Na 2 HPO 4 ; 0.3 g NH 4 Cl; 0.3g NaCl; 0.1gMgCl 2 .6H 2 O; 0.11 g CaCl 2 ; 0.5mg resazurin; 4g NaHCO 3 ; 0.25g Na 2 S.7–9H 2 O; 0.25% mucin.

[0032] Fermentation Medium 1: Brain Heart Infusion Broth (Brain Heart Infusion Powder, BHI)

[0033] Fermentation medium 2: 5g tryptone, 5g soytone, 5g beef powder, 5g yeast powder, 4g glucose, 3gNaCl, 1.5g MgSO 4 , Na 2HPO 4 , KH 2 PO 4 , water 1L, pH=7.2.

[0034] Separation and fermentation medium were autoclaved at 121°C for 15 minutes for later use.

[0035] 2. Isolation and identification of strains

[0036] ①The prepared mucin separation medium is placed in a serum bottle (filling volume 10 / 30mL)

[0037] ② Take 0.5g of feces from healthy adults and place them in sterile PBS phosphate buffered saline solution to disperse evenly. Dilute the feces in a 10-fold gradient in PBS solution in turn, and...

Embodiment 2

[0042] Embodiment 2: Akkermansia fermentation

[0043] ① Akkermansia was inoculated on brain heart powder agar with a three-section line, and cultured anaerobically at 37°C for 48 hours.

[0044] ②Pick a single colony of Akkermansia from the agar plate and inoculate it into 15mL brain heart extract powder liquid medium, and culture it anaerobically at 37°C for 24h.

[0045] ③Transfer 10mL of fermentation broth from a 15mL anaerobic tube to 200mL of brain heart extract powder liquid medium, and culture at 37°C for 24h in anaerobic conditions.

[0046] ④ 200mL of Akkermansia fermented liquid was inoculated in the brain heart extract powder liquid medium, the volume of the 5L fermentation tank was 4L, and the anaerobic gas was nitrogen and carbon dioxide mixed binary gas (N 2 :CO 2 =9:1), the stirring speed was 100r / min, the temperature was 37°C, and the anaerobic culture was carried out for 24h.

Embodiment 3

[0047] Embodiment 3: Akkermansia fermentation

[0048] ① A single colony of W03 and standard strain ATCC BAA835 (purchased from the American Type Culture Collection) was inoculated in 15 mL of fermentation medium 2, and cultured anaerobically at 37°C for 24 hours.

[0049] ②Transfer 10mL of fermentation broth from a 15mL anaerobic tube to 200mL of fermentation medium 2, and incubate at 37°C for 24 hours anaerobically.

[0050] ③ 200mL of Akkermansia fermentation broth was inoculated in fermentation medium 2, the volume of the 5L fermentation tank was 4L, and the anaerobic gas was nitrogen and carbon dioxide mixed binary gas (N 2 :CO 2 =9:1), the stirring speed is 100r / min, the temperature is 37°C, the process control pH is 6.5, and the anaerobic culture is carried out until the OD does not change.

[0051] Result: if figure 1 As shown, the W03 bacteria fermented for 13 hours with a maximum OD of 2.26 (2.13×10 9 CFU / mL) reached the stable stage, and the maximum OD of ATCC ...

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Abstract

The invention discloses a strain for preventing and treating metabolic diseases and application for the strain. The strain is Akkermansia muiniphila strain which is named Akkermansia muciniphila SSYD-3 and has the preservation number being CGMCC No.14764. Compared with the existing Ackermann muiniphila strain ATCC BAA 835, the Akkermansia muiniphila strain provided by the invention is more ideal in fermentation performance, storage stability, acid resistance, bile salt tolerance and the effect of treating the metabolic diseases.

Description

technical field [0001] The invention relates to the field of microorganisms in the field of biotechnology, in particular to a bacterial strain for preventing and treating metabolic diseases and its application. Background technique [0002] Obesity is a worldwide health problem. According to the Global Burden of Disease study, the global obesity epidemiology is gradually deteriorating, which has a negative impact on physical health and health economics. Internationally, the body mass index (BMI) is usually used as the standard to judge the degree of obesity. When 18.5≤BMI≤24.9 is in the normal range, BMI≥25 is considered overweight, and BMI≥30 is considered obese. Research data from 195 countries around the world shows that since 1980, the incidence of obesity has doubled. Currently, 5% of children and 12% of adults are obese. 1 / 5 people will be obese. From 1975 to 2014, the global average BMI of men increased from 21.7 to 24.2kg / m 2 , women increased from 22.1 to 24.4kg / ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/74A61P3/04A61P3/10A61P3/00C12R1/01
CPCA61P3/04A61P3/10A61P3/00A61K2035/11C12R2001/01C12N1/205A61K35/74A61P3/06A61K35/741A61K2035/115
Inventor 文彬宁光马新王卫庆于鸿晶洪洁孙宁云王计秋尹培军刘瑞欣高远王莎莎
Owner SHANGHAI SINE PHARMA LAB
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