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Rhizobium sp. and application thereof

A rhizobia and application technology, applied in the field of rhizobia and its application, can solve the problems that need to be strengthened, achieve the effect of no secondary risk, reduce expression elements, and enhance the therapeutic effect

Active Publication Date: 2017-01-11
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the current research on microbial envelope inhibitors still needs to be strengthened

Method used

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  • Rhizobium sp. and application thereof
  • Rhizobium sp. and application thereof
  • Rhizobium sp. and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, isolation and identification of bacterial strain NA01

[0065] 1. Isolation of strain NA01

[0066] Water samples were collected in the surface seawater of the North Atlantic Ocean, and aseptically preserved and brought back to the laboratory (Marine Ecology Laboratory, Shenzhen Graduate School, Tsinghua University) aseptically spread on a 12cm medium plate containing 2216E solid medium, and left at 30°C overnight. cultured until clearly visible single colonies emerged.

[0067] Then pick 100 single clones from the culture plate and place them in 2ml EP tubes, which contain 1ml LB liquid medium, and put these 100 single clones into a constant temperature shaker at 30°C and 220r / min for overnight culture. After the detection of the reporter strain Chromobacteriumviolaceum ATCC 12472, the positive strains that can inhibit the formation of purple circles were identified, and one of the strains was named NA01.

[0068] 2. Identification of strain NA01

[00...

Embodiment 2

[0079] Example 2, Rhizobium sp. (Rhizobium sp.) NA01 CGMCC No.12205 inhibition of pathogenic bacteria biofilm formation experiment

[0080] 1. Fermentation culture of Rhizobium sp. NA01 CGMCC No.12205 and extraction of fermentation products (metabolites)

[0081] Take out the rhizobium (Rhizobium sp.) NA01 CGMCC No.12205 obtained in Example 1 of isolation and identification from the refrigerator at -80°C, after thawing at room temperature, streak and separate on the LB solid medium plate several times, 30°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 30° C. and 220 r / min on a shaker. 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 to 36h, until the nutrients in the medium are basically exhaus...

Embodiment 3

[0092] Embodiment 3, Rhizobium sp. (Rhizobium sp.) NA01 CGMCC No.12205 inhibits pathogen virulence factor experiment

[0093] 1. Detection of the inhibitory effect of metabolites of Rhizobium sp. NA01 CGMCC No.12205 on siderophore production of Pseudomonas aeruginosa

[0094] (1) The bacterium liquid of the activated Pseudomonas aeruginosa (Pseudomonas aeruginosa) that embodiment 2 step 2 obtains is diluted to OD with LB liquid medium liquid medium 600 is 0.05.

[0095] (2) In the diluted Pseudomonas aeruginosa (Pseudomonas aeruginosa) bacterium liquid, add the filtrate supernatant (0, 1%, 3%, 5%) that the embodiment 2 step 1 of different volume fractions obtains, extract Aqueous phase (5%) and organic phase (0.5%), each concentration was repeated 3 times, placed in a shaker, 220r / min, cultured at 30°C for 16h, then centrifuged at 12000r / min for 15min at 4°C.

[0096] (3) Take three 10ml supernatants, add 20μl of three different hydrolysis substrates (siderophores) with a co...

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Abstract

The invention discloses Rhizobium sp. and application thereof. The Rhizobium sp. is preserved in CGMCC (China General Microbiological Culture Collection Center) on March 11, 2016, and the preservation number of the Rhizobium sp. is CGMCC No. 12205. The Rhizobium sp. has the advantages that the Rhizobium sp. can efficiently inhibit the formation of a pathogen biofilm and inhibit pathogen virulence factors, the fermentation products or metabolites of the Rhizobium sp. can be effectively used as the active components of pathogen biofilm inhibitors, the Rhizobium sp. and the fermentation products or metabolites thereof can be used to effectively inhibit the formation of the pathogen biofilm and control the toxicity of pathogens, and accordingly the Rhizobium sp. can be effectively used for treating bacterial infection so as to enhance the susceptibility of the pathogens to common medicine and enhance a treatment effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to rhizobia and its application. Background technique [0002] Bacterial infection has always been a major problem that threatens and plagues human health. The discovery of antibiotics represented by penicillin in the 20th century opened a new chapter for the treatment of bacterial diseases. Over the past half century, most infectious diseases have been effectively treated. But at the same time, it has also led to the abuse and irrational application of antibiotics, leading to the increase of bacterial resistance. More and more pathogenic bacteria are less sensitive to antibiotics or even disappear, and the expansion of drug resistance spectrum and the enhancement of drug resistance have brought great challenges to bacterial treatment and also pose a huge threat to public health. Recently, some superbugs, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P1/00C12R1/41
CPCA01N63/00C12N1/20C12N1/205C12R2001/41
Inventor 周进蔡中华常虹劳永民晋慧陈璐余时琛陈淑文
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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