Enterococcus faecalis resistant to cephalosporin and capable of highly expressing Sir2 like protein, and application

A technology of Enterococcus faecalis and high expression is applied in the field of new strain screening and application mechanism, which can solve the problem of no relevant reports, and achieve the effect of improving mitochondrial respiration and enhancing aerobic oxidation.

Pending Publication Date: 2020-02-18
广州溯原生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether the Sir2-like protein of Enterococcus faecalis in this study will participate in cell metabolism and act as a m...

Method used

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  • Enterococcus faecalis resistant to cephalosporin and capable of highly expressing Sir2 like protein, and application
  • Enterococcus faecalis resistant to cephalosporin and capable of highly expressing Sir2 like protein, and application
  • Enterococcus faecalis resistant to cephalosporin and capable of highly expressing Sir2 like protein, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Screening of Enterococcus faecalis resistant to cephalosporin and highly expressing Sir2 protein

[0025] 1. Experimental method materials

[0026] 1.1 Preliminary Screening of Probiotics

[0027] Isolation and culture of Enterococcus faecalis CMS-H001 seed bacteria Enterococcus faecalis CMS-H001 seed bacteria was isolated from the descending segment of the duodenum from a healthy adult. The specimens were cultured in an anaerobic box at 37°C for 4 hours and then taken out. Press 10- 1Serial dilution to 10-7, 100ul each of the stock solution and various dilutions are added dropwise to the MRS plate, the L-shaped glass rod is evenly coated, and the colony is taken out after anaerobic incubation at 37°C for 48h to observe the colony morphology. Pick a single clone for Gram staining For microscopic examination, single colonies of different morphology and staining properties were picked into MRS liquid medium, cultured in an anaerobic environment at 37°C for 24 hours, a...

Embodiment 2

[0079] Example 2 Enterococcus faecalis Enterococcus faecalis EF23 sir2 (EF-sir2) gene transfection cell and its intracellular function

[0080] 1. Experimental materials

[0081] 1. 1 Bacteria, plasmids and cells

[0082]

[0083] 1.2 Main culture medium and reagent

[0084] (1) Buffer A: consists of 150 mM sodium chloride, 500 mM Tris-HCl, 0.1% TritoX-100, pH 8.0.

[0085] (2) Buffer B: composed of 50 mM Tris-HCl, 0.1% TritonX-100, pH 8.0.

[0086] (3) Antibody diluent: 5% BSA solution, stored at 4°C for later use.

[0087] (4) Cell growth medium: Fetal bovine serum (final concentration is 10%) and penicillin streptomycin solution (final concentration is 100 units / ml) are added to the culture medium and stored at 4°C for later use.

[0088] (5) Cell maintenance solution: fetal bovine serum and penicillin streptomycin solution (final concentration of 100 units / ml) (final concentration of 2%) are added to the culture medium and stored at 4°C for later use.

[0089] (6) Trypsin solution: We...

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Abstract

The invention discloses Enterococcus faecalis resistant to cephalosporin and capable of highly expressing Sir2 like protein, and an application. The Enterococcus faecalis EF23 resistant to cephalosporin and capable of highly expressing Sir2 like protein is preserved in Wuhan university collection center of China Center for Type Culture Collection, the preservation number is CCTCC M 2018012, the preservation address is Luojia mountain, Wuchang, Wuhan city, Hubei Province, and the preservation date is 8th January, 2018. Through experiment, the inventor finds that the Enterococcus faecalis EF23 resistant to cephalosporin and capable of highly expressing Sir2 like protein can strengthen aerobic oxidation of glucose of cells, promote mitochondria respiration, and adjust and control ATP energy metabolic pathways, so that the Enterococcus faecalis EF23 resistant to cephalosporin and capable of highly expressing Sir2 like protein can treat various metabolism relevant diseases of cell senescence, mitochondria dysfunction, metabolic syndromes and the like.

Description

Technical field [0001] The present invention belongs to the field of new bacterial species screening and application mechanism. More specifically, the present invention relates to an Enterococcus faecalis which is resistant to cephalosporin and highly expresses Sir2-like protein and its application. Background technique [0002] Sir2 protein family (Sirtuin) is a kind of conserved protein, which mainly acts as NAD+ dependent protein deacetylase, which exists in various organisms including archaea to mammals. At present, many studies have shown that the mammalian Sirtuin protein family is involved in regulating cell metabolism, and helps to delay or even prevent metabolism-related diseases. [0003] Studies have shown that sirtuins are widely involved in regulating the body's energy metabolism and maintaining the body's steady state of glucose and lipid metabolism. Among them, SIRT1, SIRT3, and SIRT6 are the most widely studied. SIRT1 is the most studied member of this family. Curr...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23L33/135A23L33/195A61K35/744A61P39/06A61P3/00C12R1/01
CPCA23L33/135A23L33/195A61P39/06A61P3/00A23V2002/00A61K2035/115C12R2001/46C12N1/205A23V2200/302A23V2200/30A23V2250/546
Inventor 孙晗笑徐臻睿利时雨
Owner 广州溯原生物科技股份有限公司
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