Bifidobacterium breve NX-5 and application thereof in oxidation resistance

A technology of Bifidobacterium breve and NX-5, applied in the field of microorganisms, can solve the problems of not being suitable, few probiotics, and dependence on imports of probiotics, so as to reduce the level of ROS and improve the activity of SOD.

Inactive Publication Date: 2021-07-23
广东南芯医疗科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies and applications of probiotics in antioxidants and anti-aging
At the same time, the current international patent applications for probiotics are concentrated in the United States, Japan, and Russia, traditional research and development powerhouses, while my country lacks functional strains with independent intellectual property rights
The probiotic strains used by domestic production enterprises have long relied on imports, and foreign strains may not be suitable for the physiological conditions of the gastrointestinal tract of Chinese residents
In addition, the function of probiotics lacks strong scientific research evidence, which seriously affects the promotion of probiotics and their products

Method used

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  • Bifidobacterium breve NX-5 and application thereof in oxidation resistance
  • Bifidobacterium breve NX-5 and application thereof in oxidation resistance
  • Bifidobacterium breve NX-5 and application thereof in oxidation resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation, Identification and Preservation of Bifidobacterium breve NX-5

[0024] (1) Separation: After gradient dilution of healthy infant feces, inoculate them in BHI solid medium, BS solid medium, and MRS solid medium respectively, culture anaerobically at 37°C for 48 hours, pick a single colony on the plate and streak Pure colonies were isolated. The pure colony on the plate was inoculated in BS liquid medium, cultured anaerobically at 37°C for 12-16h, added 20% glycerol, and stored in a -80°C refrigerator.

[0025] (2) Morphological identification of bacterial strains: Gram-stained strains were observed under a microscope, Gram-positive bacteria were purple, and Gram-negative bacteria were red.

[0026] (3) Molecular biological identification of the strain: Genomic DNA was extracted from the obtained strain, and the full-length 16S rDNA fragment was amplified by PCR using 16S rDNA universal primers 27F and 1492R, followed by sequencing to identify the sp...

Embodiment 2

[0032] Preparation of embodiment 2 Bifidobacterium breve NX-5 fermentation supernatant (extracellular secretion), bacterium suspension (thalline)

[0033] Bifidobacterium breve NX-5 was activated and cultured and inoculated in BS liquid medium. After culturing at 37°C for 15 hours, the concentration of fermentation bacteria was adjusted to 1×10 6 CFU / mL, centrifuged at 4°C, 6000r / min for 10min to obtain the culture supernatant and cell pellet, the supernatant was filtered through a 0.22μm filter membrane to obtain the fermentation supernatant (extracellular secretion); the cell pellet was filtered through PBS twice After the first wash, the cells were resuspended with PBS, and the cell concentration was adjusted to 1×10 6 CFU / mL to obtain bacterial suspension (bacteria).

Embodiment 3

[0034] Example 3 Effect of Bifidobacterium breve NX-5 on ROS level in zebrafish oxidative stress model

[0035] Reduced glutathione (GSH), menadione, and dimethyl sulfoxide (DMSO) were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; 2',7'-dichlorodihydrofluorescein diacetate ( DCFH-DA) was purchased from Sigma-Aldrich Company.

[0036] The healthy wild-type AB line zebrafish developed to 4dpf (days post fertilization) was selected and placed in a 6-well cell culture plate, with 20 fish per well. The experiment set up a blank control group, a model group, a positive control group, and a sample (bacteria suspension, fermentation supernatant) intervention group, with 20 fish in each group. Add PBS to the blank control group, add PBS to the model group, add GSH solution (100μM) to the positive control group, add bacterial suspension to the bacterial suspension group, add fermentation supernatant to the fermentation supernatant group, 2.5mL per well; incubate at 28°C for 2...

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Abstract

The invention discloses bifidobacterium breve NX-5 and application thereof in oxidation resistance, and belongs to the technical field of microorganisms. The preservation number of the bifidobacterium breve NX-5 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No.20113; and fermentation supernate and bacterial suspension of the bifidobacterium breve NX-5 can remarkably reduce the level of reactive oxygen species in a zebra fish body and remarkably improve the activity of superoxide dismutase in the zebra fish body in an in-vivo oxidative stress model, and have the potential of being applied to in-vivo oxidation resistance. The bifidobacterium breve NX-5 disclosed by the invention has a huge potential application prospect in the aspect of oxidation resistance.

Description

technical field [0001] The invention relates to the technical field of microorganisms, and more specifically relates to a strain of Bifidobacterium breve NX-5 and its application in anti-oxidation. Background technique [0002] Reactive oxygen species (ROS) are by-products of cellular aerobic metabolism, mainly including hydrogen peroxide (O 2 2- ), singlet oxygen ( 1 o 2 ), superoxide radicals (O 2 ·- ), hydroxyl radicals (·OH), etc., play an important role in the cell life cycle. Low concentrations of ROS can be used as key signaling molecules in cells to regulate cell growth, proliferation and differentiation. However, the accumulation of ROS can severely damage cellular biomacromolecules such as proteins, lipids, and DNA, leading to a variety of chronic diseases, including atherosclerosis, arthritis, diabetes, neurodegenerative diseases, aging, inflammatory bowel disease, etc. Among them, ROS-mediated oxidative stress-induced senescence can be said to be the most ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L33/135A23C9/123A61K35/745A61K8/99A61P39/06A61Q19/08C12R1/01
CPCA23C9/1234A23L33/135A61K35/745A61K8/99A61Q19/08A61P39/06A23V2002/00A23V2400/519A23V2200/30
Inventor 张召陈涛
Owner 广东南芯医疗科技有限公司
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