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Stenotrophomonas maltophilia and wide-spectrum maltocin synthesized from Stenotrophomonas maltophilia and application thereof

A oligotrophomonas and maltophilia technology, applied in the field of microorganisms, can solve the problems of narrow bactericidal spectrum, inability to kill or inhibit, and achieve the effects of wide bactericidal spectrum, high bactericidal activity, good development and application prospects

Active Publication Date: 2018-09-28
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Maltophilin P28 is sensitive to high temperature and protease, and its bactericidal spectrum is narrow. It can only kill Stenotrophomonas maltophilia. Only 36 of the 80 strains tested were resistant to maltophilia P28 Sensitive and unable to kill or inhibit other strains

Method used

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  • Stenotrophomonas maltophilia and wide-spectrum maltocin synthesized from Stenotrophomonas maltophilia and application thereof
  • Stenotrophomonas maltophilia and wide-spectrum maltocin synthesized from Stenotrophomonas maltophilia and application thereof
  • Stenotrophomonas maltophilia and wide-spectrum maltocin synthesized from Stenotrophomonas maltophilia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Screening and identification process of embodiment 1 bacterial classification

[0031] 1.1 Screening of Stenotrophomonas maltophilia S16 strain

[0032] 1.1.1 Inoculate a single colony of Stenotrophomonas maltophilia into 5mL liquid LB medium, and cultivate overnight at 30°C and 200rpm;

[0033] 1.1.2 Mix 100 μL overnight culture solution with 4.5 mL semi-solid medium, pour it into LB plates to prepare double-layer plates;

[0034] 1.1.3 Transfer the overnight culture solution in step 1.1.1 to fresh liquid LB medium according to 1% inoculum amount. After culturing for 3 hours, add mitomycin C (final concentration 0.5 μg / mL) and continue culturing Overnight; take 1mL of the bacterial solution and centrifuge at 4°C, 10000rpm for 1min, take the supernatant, and filter;

[0035] 1.1.4 Take 2.5 μL of the above-mentioned culture supernatant and drop it on the double-layer plate. After standing overnight at room temperature, observe whether a transparent circle is formed on ...

Embodiment 2

[0053] The acquisition and identification of embodiment 2 maltophilin

[0054] 2.1 Obtaining maltophilin

[0055] Stenotrophomonas maltophilia was cultured in 100mL LB liquid medium at 30°C until the bacterial liquid OD 600 When the value was 0.5, mitomycin C (0.5 μg / mL) was added, and the culture was continued for 12-14 hours with shaking at 200 rpm. The cultured bacterial solution was centrifuged at 10,000 g for 10 min at 4°C, the supernatant was aspirated, PEG8000 (4%, m / v) and NaCl (3%, m / v) were added, mixed well, and left standing overnight at 4°C. Centrifuge at 10,000 g for 30 min at 4°C, suspend the precipitate in 1 mL of TE buffer, and obtain the maltophilin extract after filtration.

[0056] The maltophilin crude extract was loaded on a DEAE-cellulose ion-exchange chromatography column, and sequentially dissolved in 0.01M phosphate buffer (pH6.8) with a nonlinear gradient NaCl solution (0, 0.3, 0.4 and 0.7 M) eluting to obtain a purified sample.

[0057] 2.2 Malt...

Embodiment 3

[0060] Example 3 Variation of the bactericidal activity of maltophilin at different temperatures

[0061] Incubate the samples of maltophilin S16 at different temperatures for 10 min, and then quickly cool them in ice, then dilute the samples with 2-fold gradient with TE buffer solution, take 2.5 μL of the diluted solution and drop them on the double-layer plate in turn, and incubate at 30 °C Then observe the inhibition zone and calculate the bactericidal activity of maltophilin.

[0062] Table 2 Residual bactericidal activity of maltophilin after treatment at different temperatures (×400AU / mL)

[0063]

[0064]

[0065] Note: Maltophilia 16 indicator strain is Stenotrophomonas maltophilia S21 strain;

[0066] The indicator strain of maltophilia P28 is Stenotrophomonas maltophilia strain c6.

[0067]The results in Table 2 show that when the temperature is 45°C or below, the bactericidal activity of maltophilin S16 has no effect. When the temperature rises to 50°C, its ...

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Abstract

The invention belongs to the technical field of microbes and specifically relates to Stenotrophomonas maltophilia and wide-spectrum maltocin synthesized from Stenotrophomonas maltophilia and application thereof. The Stenotrophomonas maltophilia is named Stenotrophomonas maltophilia S16 strain and was preserved in the China Center for Type Culture Collection with the preservation number being CCTCCM 2017431. The Stenotrophomonas maltophilia S16 strain is a new strain, and the strain can be induced by mitomycin C to produce maltocin S16. The maltocin S16 has high bactericidal activity and widebactericidal spectrum, can kill most Stenotrophomonas maltophilia as well as Escherichia coli, has resistance to protease, is relatively stable at high temperature, and has a good development and application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a strain of Stenotrophomonas maltophilia and the broad-spectrum maltocin synthesized by it and its application. Background technique [0002] In recent years, with the irrational use of antibiotics, multi-drug resistant organisms (MDRO) are increasing. The test results of the sensitivity and drug resistance of clinical isolates in major areas of China to commonly used antimicrobials show that Escherichia coli (Escherichia coli) is the most clinically isolated strains, reaching about 20%, of which more than half of Escherichia coli produce extended-spectrum β- Lactamase (Extended-Spectrum β-Lactamase, ESBL). It was found that the resistance rates of Escherichia coli to ciprofloxacin, gentamicin, piperacillin and trimethoprim-sulfamethoxazole were close to or higher than 50%. At the same time, Stenotrophomonas maltophilia, which is resistant to multiple antibiotics, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C07K14/195A61K38/16A61P31/04C12R1/01
CPCA61K38/00C07K14/195C12N1/205C12R2001/01Y02A50/30
Inventor 黄玉屏陈静仪朱尧
Owner WUHAN UNIV
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