A sixteen-carbon chain fatty acid antagonist produced by Bacillus amyloliquefaciens sqr9 and its application

A technology of amylolytic spores and fatty acids, which is applied in the field of microorganisms, and can solve the problems of poor efficacy, ineffectiveness, and complicated bacterial resistance of antibiotics, etc.

Active Publication Date: 2020-03-17
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Overuse of antibiotics causes bacteria to gradually produce drug-resistant genes, making antibiotics less effective or even ineffective
Moreover, bacterial resistance can be transmitted to each other, complicating bacterial resistance, so it is urgent to find new antibiotics to solve the current problems

Method used

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  • A sixteen-carbon chain fatty acid antagonist produced by Bacillus amyloliquefaciens sqr9 and its application
  • A sixteen-carbon chain fatty acid antagonist produced by Bacillus amyloliquefaciens sqr9 and its application
  • A sixteen-carbon chain fatty acid antagonist produced by Bacillus amyloliquefaciens sqr9 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: verify the growth inhibitory activity of fatty acid of the present invention to homologous bacterial strain:

[0024] The gene island GI3 of Bacillus amyloliquefaciens SQR9 is a horizontal gene transfer gene island, located at position 657050-738610 in the nucleotide sequence of the SQR9 genome. The gene contained in GI3 does not exist at the same position in the homologous strain. It is inferred that the gene island GI3 is synthesized The active substances are related to homologous strains. To investigate the function of the gene island GI3 in SQR9, the gene island was fully knocked out by double crossover homologous recombination. First, the gene of the chloramphenicol (Cm) resistance selection marker was fused with the upstream and downstream homology arms of the gene to be knocked out by fusion PCR method, and the fusion fragment was transferred into competent SQR9, 37°C, 100r min -1 After recovering and culturing for 8 hours, spread the bacterial sus...

Embodiment 2

[0029] Example 2: Preparation and separation and purification of the fatty acid antagonistic substances of the present invention:

[0030] The mutant △GI3 only knocks out the gene island GI3, so that the mutant △GI3 can no longer synthesize the sixteen-carbon chain fatty acids mentioned in the application, that is, C 17 h 34 o 3 , without affecting the synthesis of other active substances in SQR9, so the difference between the experimental group and the control group is C 17 h 34 o 3 biological activity. In Example 1, the growth inhibitory effect of the mutant △GI3 on ​​FZB42 completely disappeared, so we can separate and purify the sixteen-carbon chain fatty acids synthesized by △GI3 through activity tracking, that is, each step of separation verifies the effect on FZB42. Growth inhibitory activity, active components are further separated and purified.

[0031] First prepare the seed solution: pick a single colony of SQR9 and put it in a test tube of Landy medium, 170 rpm...

Embodiment 3

[0035] Embodiment 3: verify the growth inhibitory activity of fatty acid of the present invention to plant pathogenic bacteria:

[0036] Seven strains of pathogenic fungi and one strain of pathogenic bacteria were selected to verify the antagonistic activity of the isolated long-chain saturated fatty acid solution (100 μg / ml). The seven selected pathogenic bacteria are:

[0037] Cucumber Fusarium oxysporum (Fusarium oxysporum.sp.cucumebrium Owen, isolated in our laboratory),

[0038] Sclerotias clerotiorum (Sclerotias clerotiorum, isolated in our laboratory),

[0039] Banana Fusarium oxysporum Schl (Fusarium oxysporum Schl, isolated in our laboratory),

[0040] Phytophthora capsicum (Phytophthora capsici, ACCC NO.36278),

[0041] Rhizoctonia solani Ktihn (ACCC NO.36246),

[0042] Rhizoctonia graminearum / Wheat sheath blight (Rhizoctonia cerealis, ACCC NO.37393),

[0043] Corn stalk rot Fusarium Verticillioides (isolated in our laboratory)

[0044] Ralstonia solanacearum (...

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Abstract

The invention discloses a hexadecenoic chain fatty acid agonist substance generated from bacillus amyloliquefaciens SQR9 and application thereof. The molecular formula of the hexadecenoic chain fatty acid agonist substance is C17H34O3, and the hexadecenoic chain fatty acid agonist substance has the advantages of wide antibacterial spectrum, good growth inhibitory activity for plant pathogenic fungi and pathogenic bacteria, obvious growth inhibitory effect on pathogenic bacteria and pathogenic fungi, safety, stability and a wide application prospect and can be applied to the inhibition of the growth of the plant pathogenic fungi and pathogenic bacteria.

Description

technical field [0001] The invention belongs to the field of microorganisms, in particular to a sixteen-carbon chain fatty acid antagonist produced by Bacillus amyloliquefaciens SQR9 and an application thereof. [0002] technical background [0003] Antibiotics are a class of secondary metabolites with anti-pathogenic or other activities produced by microorganisms including bacteria, fungi, actinomycetes, or higher animals and plants during their life. The overuse of antibiotics causes bacteria to gradually produce drug-resistant genes, making antibiotics less effective or even ineffective. Moreover, bacterial drug resistance can be transmitted to each other, complicating bacterial drug resistance, so it is urgent to find new antibiotics to solve the current problems. The traditional method of searching for new antibiotics is to cultivate newly isolated strains under different conditions, then isolate and purify the fermentation products, and finally identify new antibiotics...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C59/01C12P7/42A01N37/36A01P1/00A01P3/00C12P7/64C12R1/07
CPCA01N37/36C07C59/01C12P7/42C12P7/6409
Inventor 张瑞福王丹丹徐志辉张桂山邵佳慧沈其荣
Owner NANJING AGRICULTURAL UNIVERSITY
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