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Microbial inhibitor

A technology of microorganisms and inhibitors, applied in the field of microbial inhibitors, which can solve the problems of undeveloped biological functions and low yields

Inactive Publication Date: 2012-06-27
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Deoxyviobacterin and oxidized violacein are derivatives of violacein, by-products in the biosynthesis process, with low yields, and no research has been carried out on their biological functions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, preparation and analysis of extract A, extract B and extract C

[0059] C. freundii (pCOM10) strain, B2 strain and C. freundii (pCOMΔD) strain violacein substances (including violacein or violacein derivatives) synthesized by the three strains are not secreted extracellularly, so violacein The extraction of the pigment substances all includes the following steps: fermentation, centrifugation to harvest the thallus, and freezing and thawing to break the cell wall of the thallus to release the pigment. The violacein substances are insoluble in water and easily soluble in organic solvents. Ethanol is the best solvent for extracting violacein substances. Therefore, absolute ethanol is used to extract violacein substances in broken cells.

[0060] 1. Preparation of Extract A

[0061] The liquid fermentation medium is obtained by distilling the following solutes to 100mL with water: NaH 2 PO 4 2H 2 O 0.13g, Na 2 HPO 4 12H 2 O 0.3g, NH 4 Cl 0.089g, KH 2 ...

Embodiment 2

[0141] Embodiment 2, extract A, extract B and extract C are to the inhibitory effect of test bacterium

[0142] Detect respectively the violacein substances (extract A, extract B or extract C) prepared by Example 1 to the inhibition of 10 strains of test bacteria, the specific steps are as follows:

[0143] 1. Preparation of bacteria-containing plates

[0144] 100 μL of bacterial culture solution (OD 600 =0.1) was applied to a plate containing NA solid medium, and left to dry to make a plate containing bacteria.

[0145] 2. The inhibitory effect of violacein substances on the tested bacteria (aseptic operation)

[0146] ① Dissolve Extract A, Extract B, and Extract C in acetone, respectively, so that the concentration is 125 μg / mL.

[0147] ②Put 3 groups of sterile filter paper disks with a diameter of 5mm (3 pieces are 1 group) in the three solutions obtained in step ① and soak for 12 hours, then air-dry to remove acetone; put 1 group of the same sterile filter paper disks ...

Embodiment 3

[0164] Embodiment 3, extract A, extract B and extract C are to the inhibitory effect of testing fungus

[0165] 1. The inhibitory effect of violacein substances on the tested fungi

[0166] Detect respectively the violacein substances (extract A, extract B or extract C) prepared in Example 1 to the inhibition of 24 strains of tested fungi, the specific steps are as follows:

[0167] 1. Determination of the sensitivity of violacein derivatives to the inhibition of pathogenic fungi (aseptic operation)

[0168] ① Dissolve Extract A, Extract B and Extract C in sterile anhydrous acetone, then add an equal volume of water, and dilute the mother liquor with a concentration of 20 mg / mL with 50% acetone aqueous solution.

[0169] ② Add the three mother liquor solutions in step ① to different sterilized plates, then pour into the PDA solid medium that has been melted and cooled to about 45°C, so that the final concentration of the extract is 2 mg / mL, and mix well , placed to dry to ma...

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Abstract

The invention discloses a microbial inhibitor. The microbial inhibitor provided in the invention is violacein substances (comprising violacein or violacein derivatives) which are extracted from Citrobacter freundii (pCOM10) strains, Citrobacter freundii (pCOMdeltaD) strains or Duganella B2 and have different chemical structures. Two of three extracts provided in the invention have strong inhibition on various plant causative fungi and gram positive bacteria. The extract from the Citrobacter freundii (pCOM10) strains can inhibit the various plant causative fungi, and the inhibition rate of the Citrobacter freundii (pCOM10) strains with the concentration of 4mg / mL on Phytophthora capsici can reach 100%. The microbial inhibitor establishes bases for developing agricultural antibiotics through utilizing violacein or the violacein derivatives.

Description

technical field [0001] The present invention relates to a microbial inhibitor. Background technique [0002] Chemical pesticides are highly polluting, and long-term use poses a safety hazard to human health. Biopesticides not only have high safety and strong selectivity, but also have the characteristics of easy degradation and high activity, and have attracted widespread attention from all over the world. Researchers have found that agricultural antibiotics derived from microorganisms have become an important substitute for chemical pesticides, and thus become a research hotspot for researchers from all over the world. [0003] Violacein is an indole derivative synthesized by bacteria using L-tryptophan as a precursor. The biosynthetic pathway of violacein has been basically clarified. All the C, H, and N atoms in violacein come from tryptophan molecules. The whole synthetic pathway of violacein involves 5 genes vioabcde, which are distributed in the same transcription ...

Claims

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Application Information

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IPC IPC(8): A01N43/38A01P1/00A01P3/00A01N63/02
Inventor 邢新会王海胜闫艳春
Owner TSINGHUA UNIV
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