Application of aminopeptidase reducing function

A technology of aminopeptidase and aminopeptidase coding, applied in the application field of enhancing the susceptibility of bacteria to antibiotics, can solve problems such as food pollution, fluid flow resistance, material surface corrosion, etc., to promote disintegration and weaken drug resistance Effect

Inactive Publication Date: 2018-02-09
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When microorganisms form a biofilm on the surface of the medium, it usually causes material surface corrosion, fluid flow resistance, pipeline blockage, continuous release of harmful bacteria, etc., resulting in energy waste and food pollution, etc.

Method used

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  • Application of aminopeptidase reducing function
  • Application of aminopeptidase reducing function
  • Application of aminopeptidase reducing function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0031] According to another preferred embodiment of the present invention, said reducing the function of aminopeptidase includes reducing the extracellular secretion of aminopeptidase.

[0032] In addition to reducing the expression of aminopeptidase so as to reduce its extracellular secretion in the above way, the following methods can also be used, for example, to reduce or inhibit the function of aminopeptidase-dependent extracellular secretion signal peptide, so that aminopeptidase Losing the guidance of the extracellular secretion signal peptide or reducing the guidance function of the extracellular secretion signal, so that the amount of aminopeptidase outside the cell is greatly reduced, and the biofilm loses its dependence on its formation to hinder the formation of the biofilm or disintegrate the biofilm .

[0033] Wherein, the method for reducing or inhibiting the function of the aminopeptidase-dependent extracellular secretion signal peptide may include knockout, in...

Embodiment 1

[0060] This example is used to illustrate the effect of PaAP deletion on the growth of Pseudomonas aeruginosa

[0061] 1. Inoculate Pseudomonas aeruginosa PAO1 and PaAP deletion mutant ΔpaaP on LB solid medium plate, and culture overnight at 37°C.

[0062] 2. Pick out the single clones of PAO1 and ΔpaaP from the plate that completed step 1, inoculate them into LB liquid medium, and cultivate overnight at 37°C and 200rpm with shaking.

[0063] 3. After completing step 2, inoculate 1ml of the bacterial solution into 100ml of Jensen’s medium, culture at 37°C with shaking at 200rpm, and use a spectrophotometer to detect the absorbance of the bacterial solution at 600nm at different times. see results figure 1 .

[0064] Depend on figure 1 It can be seen that the OD of PAO1 and ΔpaaP at different times 600 There was no significant difference, indicating that the deletion of PaAP had no significant effect on the growth of Pseudomonas aeruginosa.

Embodiment 2

[0066] This example is used to illustrate the effect of PaAP deletion on biofilm disintegration

[0067] 1. Inoculate Pseudomonas aeruginosa PAO1, PaAP deletion mutant ΔpaaP, PaAP complementation strain ΔpaaP::PaAP, PaAP point mutant ΔpaaP::D308A, and signal peptide-deleted PaAP mutant ΔpaaP::PaAPNS on LB solid culture Incubate overnight at 37°C on base plates.

[0068] 2. Same as 2 of Implementation Case 1.

[0069] 3. Inoculate 1 μl of the overnight bacterial solution into 100 μl of Jensen’s medium, add it to a 96-well PVC plate (Falcon3911), and culture at 30°C for 12h, 24h, 36h and 48h.

[0070] 4. The biomass of bacteria adsorbed on the 96-well plate was detected by the crystal violet method. (1) Suck away the free and loosely adsorbed bacteria with a row gun, and gently wash 3 times with normal saline. (2) Add 120 μL of 0.1% crystal violet to stain the firmly adsorbed bacterial cells in the small holes, and let stand at 30° C. for 10 minutes. (3) Take out the PVC boa...

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Abstract

The invention relates to the field of biology, in particular to application of an aminopeptidase reducing function in precaution and/or prevention of thallus biofilm formation, promotion of biofilm degradation and PslG release, reduction of the thallus content in a biofilm and improvement of antibiotic sensibility of thalli and application of an aminopeptidase reducing function method in preparation of drugs for precaution and/or prevention of thallus biofilm formation, promotion of biofilm degradation and PslG release, reduction of the thallus content in a biofilm and improvement of antibiotic sensibility of thalli. By adopting the aminopeptidase reducing function, lots of thalli in the biofilm are dead due to nutrition deficiency, dead bacteria release Psl polysaccharide hydrolase, hydrolysis of exopolysaccharides in the biofilm is promoted, and finally biofilm disruption is caused. Therefore, by developing drugs for aminopeptidase, biofilm disruption is promoted, the drug resistanceof biofilm thalli is reduced, and finally the purpose of removing harmful bacterium biofilms with antibiotics is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to reducing the function of aminopeptidase in preventing and / or preventing the formation of bacterial biofilms, promoting biofilm degradation and the release of PslG (exopolysaccharide hydrolase), and reducing the content of bacterial cells in biofilms And the application in enhancing the susceptibility of bacterial cells to antibiotics; and the method for reducing aminopeptidase function is used in the preparation for preventing and / or preventing bacterial cell biofilm formation, promoting biofilm degradation and the release of PslG, reducing biofilm internal bacterial cells content and the application of drugs that enhance the susceptibility of bacteria to antibiotics. Background technique [0002] Biofilm refers to a highly organized and systematic multicellular population structure formed by microorganisms growing on the surface of a carrier through adhesion and wrapping with their ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61P31/04
CPCA61K45/00A61K48/00
Inventor 马旅雁赵天湖王世伟
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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