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Proteus mirabilis and application thereof in suppression of biofilm and detoxification

A technology of proteus mirabilis and biofilm, applied in the field of proteus mirabilis, can solve the problems of bacterial drug resistance and reduce the effect of antibiotic treatment, and achieve the effect of reducing virulence and low probability of drug resistance

Active Publication Date: 2013-02-06
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the frequent use of antibiotics has caused bacteria to develop increasingly serious drug resistance, greatly reducing the therapeutic effect of antibiotics

Method used

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  • Proteus mirabilis and application thereof in suppression of biofilm and detoxification
  • Proteus mirabilis and application thereof in suppression of biofilm and detoxification
  • Proteus mirabilis and application thereof in suppression of biofilm and detoxification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] The isolation and identification of embodiment 1, bacterial strain 10#

[0068] One, the isolation of bacterial strain 10#

[0069] In the pond of Shenzhen Graduate School of Tsinghua University, put several disposable plastic plates. Take out 3 petri dishes every day, wash 3 times with sterile water, smear the petri dish with sterile cotton swab dipped in sterile water, and then spread the cotton swab on a 12cm medium plate containing LB solid medium, and culture overnight at 30°C , until clearly visible monoclonal growth.

[0070] Then pick 100 monoclonals from the culture plate and put them in 2ml EP tubes containing 1ml LB liquid medium. Put these 100 tubes of monoclonals into a constant temperature shaker at 30°C and 220r / min for overnight culture. One of the bacterial strains is named as 10# bacteria.

[0071] 2. Identification of strain 10# bacteria

[0072] The strain 10# obtained in step 1 was identified from the aspects of morphology, physiological and bio...

Embodiment 2

[0083] Example 2. Detection of Proteus mirabilis 10#CGMCC No.6426 inhibiting pathogenic bacteria biofilm

[0084] 1. Fermentation culture of Proteus mirabilis 10#CGMCC No.6426 and extraction of fermentation products (metabolites)

[0085]Take out the preserved Proteus mirabilis (Proteusmirabilis) 10#CGMCC No.6426 isolated and identified in Example 1 from the refrigerator at -80°C, thaw at room temperature, streak and separate on LB solid medium plate for many times, and keep at 37°C Cultured for more than 12 hours, until clearly visible monoclonal growth. Three single clones were picked out and cultured overnight in 1 ml of LB liquid medium at 37° C. on a shaker at 220 r / min. Take 1ml of the bacterial solution and add it to a 250ml Erlenmeyer flask containing 100ml of LB liquid medium, put it on a shaker, 30°C, 220r / min (the radius of the centrifugal rotor is 13.5cm) and cultivate for 24h~36h, until the nutrients in the medium are basically exhausted , until the cell density...

Embodiment 3

[0095] Example 3, Detection of Proteus mirabilis 10#CGMCC No.6426 Inhibiting Pathogenic Bacteria Virulence Factors

[0096] 1. Detection of the inhibitory effect of metabolites of Proteus mirabilis 10#CGMCC No.6426 on elastase of Pseudomonas aeruginosa

[0097] (1) Dilute the activated Pseudomonas aeruginosa (Pseudomonas aeruginosa) bacterial solution obtained in step 2 of Example 2 to OD with LB liquid medium 600 is 0.05.

[0098] (2) Add different volume fractions of the filtrate (0, 3%, 5%) obtained in Step 1 of Example 2 to the diluted Pseudomonas aeruginosa bacterial solution, repeat each concentration 3 times, and put Into a shaker, 220r / min, 37 ° C for 21 hours, 4 ° C, 12000g / min, centrifuged for 5 minutes.

[0099] (3) Take 500 μl centrifuged supernatant and add 1 mL elastin buffer, shake and co-culture for 6 hours, add 600 μl 0.1M EDTA to stop the reaction, centrifuge to remove the precipitate (10000r / min, 2 minutes), measure OD 495 . The experiment was repeated t...

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Abstract

The invention discloses a bacterial isolates relative to suppression of a biofilm and detoxification. The bacterial isolates relative to suppression of the biofilm and detoxification, which is provided by the invention, is especially Proteus mirabilis (Proteus mirabilis)10#, wherein the preservation number of Proteus mirabilis10# in China General Microbiological Culture Collection Center is CGMCC No.6426. The Proteus mirabilis10# CGMCC No.6426, which is provided by the invention, has effects of inhibiting formation of the pathogenic bacteria biofilm and detoxifying. The invention has the following advantages that: (1) through active ingredients (metabolites) of a Proteus mirabilis10# CGMCC No.6426 preparation, growth of the pathogenic bacteria is not influenced, the force is not generated, and the tolerance probability is low; and (2) through active ingredients (metabolites) of a Proteus mirabilis10# CGMCC No.6426 preparation, formation of the pathogenic bacteria biofilm is inhibited and the toxicity of the pathogenic bacteria is reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a bacterial strain related to biofilm inhibition and attenuation, in particular to a Proteus mirabilis related to biofilm inhibition and attenuation and application thereof. Background technique [0002] As we all know, in today's biological and medical fields, the main method to fight against bacterial invasion and infection is to use antibiotics. However, the frequent use of antibiotics has caused bacteria to develop increasingly serious drug resistance, greatly reducing the therapeutic effect of antibiotics. More recently, there have been "superbugs" that are completely immune to antibiotics. This undoubtedly sounded the alarm for the health of human beings, animals and plants. Therefore, finding new ways to treat bacterial infections has become an urgent need. [0003] Studies have shown that the reasons for bacterial drug resistance mainly come from two aspects: on the one hand,...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04A61K35/74A61P31/04C12R1/01
Inventor 蔡中华余时琛陈璐常虹朱小山周进
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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