A kind of Bacillus subtilis with cephalosporin resistance and high expression sir2 protein and application thereof

A Bacillus subtilis, high expression technology, applied in the field of screening and application mechanism in the new army, can solve the problem of no related research, and achieve the effect of improving mitochondrial respiration and enhancing aerobic oxidation

Active Publication Date: 2020-11-17
GUANGZHOU HOPBIOYOUTH BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So whether the Sir2 protein of probiotics is involved in cell metabolism and acts as a molecular target for pr

Method used

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  • A kind of Bacillus subtilis with cephalosporin resistance and high expression sir2 protein and application thereof
  • A kind of Bacillus subtilis with cephalosporin resistance and high expression sir2 protein and application thereof
  • A kind of Bacillus subtilis with cephalosporin resistance and high expression sir2 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Experimental methods and materials

[0030] 1.1 Preliminary screening of probiotics

[0031] Collect 1.0g of feces samples from 3 young patients in Nanyang People's Hospital, amplify and culture with LB medium, remove plasmid with ethidium bromide method, dilute with distilled water to 10-8, a total of 9 dilutions, take 1mL, and spread Put it on the LB solid medium, place it in the incubator, and cultivate it for 48 hours at 37 hours. After the culture is over, take out the plate and pick out rough and opaque surface, stained white or slightly yellow colonies, and transfer to LB liquid medium for 48 hours at 37°C, and store in a refrigerator at 4°C for later use.

[0032] 1.2 Morphological observation of strains.

[0033] Inoculate the pure culture of isolated and purified strains into the enrichment liquid medium, revive after 37 hours of cultivation for 24 hours, and perform Gram staining, observe the morphological characteristics of the cells under an optical mi...

Embodiment 2

[0093] Example 2 Bacillus subtilis BS16sir2 (BS-sir2) gene transfected cells and intracellular function research

[0094] 1. Experimental materials

[0095] 1.1 Bacteria, plasmids and cells

[0096]

[0097] 1.2 Main media and reagents

[0098] (1) Buffer A: 150 mM NaCl, 500 mM Tris-HCl, 0.1% TritoX-100, pH 8.0.

[0099] (2) Buffer B: Consists of 50 mM Tris-HCl, 0.1% TritonX-100, pH 8.0.

[0100] (3) Antibody diluent: Weigh 10 g of BSA powder and dissolve it in 200 mL of TBST buffer to form a 5% BSA solution, and store it at 4°C for later use.

[0101] (4) Cell growth medium: fetal bovine serum (final concentration: 10%) and penicillin-streptomycin solution (final concentration: 100 units / ml) were added to the culture medium, and stored at 4°C for later use.

[0102] (5) Cell maintenance solution: Fetal bovine serum and penicillin-streptomycin solution (final concentration: 100 units / mL) (final concentration: 2%) were added to the culture medium and stored at 4°C for la...

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Abstract

The invention discloses a Bacillus subtilis BS16 with cephalosporin resistance and high expression of Sir2 protein and application thereof. The Bacillus subtilis BS16 with cephalosporin resistance and high expression of Sir2 protein of the present invention is preserved in the Guangdong Microbial Culture Collection Center, the preservation number is GDMCC No.60193, and the preservation address is Building 59, Compound No. 100, Xianlie Middle Road, Guangzhou City 5th floor, the preservation date is June 2, 2017. The present invention finds through experiments that Bacillus subtilis BS16, which has cephalosporin resistance and highly expresses Sir2 protein, can enhance the aerobic oxidation of glucose in cells, improve mitochondrial respiration, and regulate the ATP energy metabolism pathway. Therefore, Bacillus subtilis BS16 It can realize the treatment of various metabolic related diseases such as cell aging, mitochondrial dysfunction and metabolic syndrome.

Description

technical field [0001] The invention belongs to the field of screening and application mechanism in the new army, more specifically, the invention relates to a Bacillus subtilis BS16 with cephalosporin resistance and high expression of Sir2 protein and its application. Background technique [0002] The Sir2 protein family (Sirtuin) is a conserved protein, which mainly acts as an NAD+-dependent protein deacetylase, which exists in various organisms including archaea to mammals. Many current studies have shown that the mammalian Sirtuin protein family is involved in the regulation of cell metabolism, and helps to delay or even prevent metabolic related diseases. In addition, more and more studies have shown that probiotics can improve metabolic diseases and so on. Probiotics also have Sir2 homologous proteins, but there are few reports on the biological functions of sir2 proteins in probiotics. [0003] Studies have shown that sirtuins are widely involved in regulating the b...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P39/06A61P3/00A23L33/135C12R1/125
CPCA23V2002/00A61K2035/115A23L33/135C12N1/205C12R2001/125A23V2200/302A23V2200/30
Inventor 孙晗笑郭晴谢亚杰利时雨刘梦鸽陈露平黄金群
Owner GUANGZHOU HOPBIOYOUTH BIOTECH CO LTD
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