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Method for promoting expression of deoxyviolacein in chromobacterium violaceum

A technology of Chromobacterium violaceum and violetobacterin, which is applied in the field of biomedicine, can solve the problems of inability to realize industrialized production and low yield, and achieve the effect of facilitating promotion and promoting expression.

Active Publication Date: 2020-01-03
DALIAN NATIONALITIES UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current yields of these two blue-purple microbial metabolites are extremely low and cannot be industrialized
Therefore, there is an urgent need for a method to promote the expression of violacein and deoxyviobacterin in Chromobacter violaceum to solve the problem of low yield

Method used

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  • Method for promoting expression of deoxyviolacein in chromobacterium violaceum
  • Method for promoting expression of deoxyviolacein in chromobacterium violaceum
  • Method for promoting expression of deoxyviolacein in chromobacterium violaceum

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Embodiment 1

[0030] The synthesis of embodiment 1 histidine fatty aldehyde Schiff base

[0031] In a round-bottomed flask filled with 60 mL of absolute ethanol, add 10 mmol of histidine and 10 mmol of potassium hydroxide, stir and dissolve at 50°C under nitrogen protection. Dissolve 20mmol of aliphatic aldehyde in 20mL of absolute ethanol, slowly drop it into a round bottom flask, and stir at 55°C for 3h. After the reaction, cool to room temperature, and remove unreacted histidine by suction filtration. The filtrate solution was evaporated under reduced pressure (40°C) to give a pale yellow solid. The product was placed in a 20 mL centrifuge tube, 15 mL of acetone was added, ultrasonically cleaned for 10 minutes, centrifuged at 7000 rpm / min for 3 minutes, and washed 10 times in total to obtain a light yellow solid, which was dried in vacuo to obtain a light yellow solid. Its structure was characterized by H NMR spectroscopy. figure 1 , figure 2 , image 3 and Figure 4 shown.

Embodiment 2

[0032] The cultivation of embodiment 2 Chromobacterium violaceum

[0033] #802 medium: polypeptone 10g, yeast powder 2g, MgSO 4 ·7H 2 Add 1 g of deionized water to make up to 1 liter, and sterilize at 120°C for 30 minutes.

[0034] In a clean bench, take a sterile 24-well plate. Add 1980 μL of #802 liquid medium to each well, 20 μL of Chromobacter violaceum solution activated for 12 hours, and incubate at 30°C for 36 hours.

Embodiment 3

[0035] The identification of embodiment 3 deoxyviolet bactin

[0036] Transfer the culture solution of Example 1 to a centrifuge tube, centrifuge for 20 minutes (10000r / min) to discard the upper layer, add deionized water for ultrasonic washing for 10 minutes, and centrifuge for 20 minutes (10000r / min) to discard the upper layer. Add 1mL of methanol to the centrifuge tube for extraction, centrifuge for 20 minutes (10000r / min) to get the upper layer liquid, and evaporate to dryness on a rotary evaporator. Add 1 mL of methanol again for extraction and spin dry, and repeat the operation three times.

[0037] After identification by mass spectrometry, the extracted blue-purple pigment was deoxyviotropin with a molecular weight of 326 ( Figure 5 ).

[0038] The extracted deoxyviotropin was formulated into a methanol solution with a concentration of 1 mg / mL, and a full-wavelength ultraviolet scan was carried out with a cuvette of 200 μL, and its characteristic absorption peak was...

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Abstract

The invention relates to a method for promoting the expression of deoxyviolacein in chromobacterium violaceum, and belongs to the field of biomedicine. According to the main technical scheme, histidine fatty aldehyde Schiff base of certain concentration and the chromobacterium violaceum which is activated for 12 hours are co-cultured for 12-48 hours at 25-35 DEG C, and then the deoxyviolacein expressed by thalluses is collected. The histidine fatty aldehyde Schiff base has a five-membered ring head structure and a long-chain fatty aldehyde tail part which are very similar to a signal moleculen-hexane-L-homoserinelactone of a chromobacterium violaceum quorum sensing system, the expression of the deoxyviolacein in the chromobacterium violaceum can be effectively promoted, the method is simple, easy to implement, and convenient to popularize, and theoretical reference for solving the problem of extremely low yield of the deoxyviolacein is provided.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for promoting the expression of deoxyviotropin in Chromobacterium violaceum. Background technique [0002] Violacein is a metabolite produced by microorganisms, which belongs to indole derivatives and is a water-insoluble blue-black pigment. In the 19th century, it was discovered that Chromobacterium violaceum could express violacein, and since then, different strains have been found to be able to produce violacein. Later, it was discovered that in addition to violacein, there is another substance with a small content in this blue-purple pigment-deoxyviotropin. [0003] [0004] Studies have shown that violacein has very excellent biological activities, including broad-spectrum antibacterial properties (effectively inhibiting Streptococcus, Bacillus, Staphylococcus aureus, Pseudomonas, etc.); Anti-herpes virus, polio virus has resistance activity); anti-tumor ...

Claims

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Application Information

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IPC IPC(8): C12P17/16C12N1/38C07D233/64C12R1/01
CPCC12P17/165C12N1/38C07D233/64Y02A50/30
Inventor 张丽影权春善李容庆马浩迪石荟向航宇
Owner DALIAN NATIONALITIES UNIVERSITY
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