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Alga inhibiting active substance violacein and preparation method thereof

A technology of purple bacteriocin and algae-inhibiting activity is applied in the field of purple bacteriocin, a substance with potent algae-inhibiting activity and its preparation, which can solve the problems of high cost, secondary environmental pollution, difficult removal of chemical substances and the like

Active Publication Date: 2015-05-20
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chemical method has the advantages of simple operation and quick effect, but the cost is high and the chemical substances are difficult to remove, especially the chemical agents used will cause secondary pollution to the environment

Method used

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  • Alga inhibiting active substance violacein and preparation method thereof
  • Alga inhibiting active substance violacein and preparation method thereof
  • Alga inhibiting active substance violacein and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: Separation and screening of anti-algae bacterial strains

[0051] 1) Take a sample of river water from the Jiulong River Basin in Fujian and dissolve it in 90 mL of high-pressure sterilized Gaoshi No. I medium (2 g of soluble starch, KNO 3 0.1g, K 2 HPO 4 0.05g, MgSO 4 ·7H 2 O 0.05g, NaCl 0.05g, FeSO 4 ·7H 2 O 0.001g, distilled water 100mL, pH7.2~7.4), placed on a shaker at 150~200rpm for 20~30min to disperse the sample evenly;

[0052] 2) 10-fold dilution method, spread evenly dispersed samples on the Gaoshi No. 1 solid plate, and incubate at 27°C for 3-5 days;

[0053] 3) Pick different types of single colonies and streak them on the Gaoshi No. 1 solid plate, culture them at 27°C for 3 to 5 days, verify whether they are pure cultures, and repeat this step until pure cultures are obtained;

[0054] 4) Inoculate a single colony of the isolated pure culture into 4 mL of Goose No. 1 liquid medium, place on a shaker at 27°C, and shake at 180 rpm for 3 ...

Embodiment 2

[0056] Embodiment 2: Algae inhibiting effect assay method

[0057] 1) Heterocurvium akashiwo at 20±1°C, 12h light, 12h dark, 50μmol photons m -2 the s -1 Under the condition of light intensity, culture in the Erlenmeyer flask until the exponential phase, then divide into 24-well cell culture plate, fill each well with 2mL of algae cell suspension, and adapt to the growth for 1 day;

[0058] 2) Quantitatively add the sample to be tested into a 24-well plate;

[0059] 3) Take Heterocurvium akashiwo culture medium at regular intervals, put 200 μL samples in a 24-well plate, and use a microplate reader to detect the fluorescence intensity at 680 nm under the excitation of 440 nm excitation light, according to the measured fluorescence intensity of algal cells in the control group and the treatment group At the same time, the morphological changes of algal cells were observed.

Embodiment 3

[0060] Embodiment 3: Separation and identification of algae-inhibiting active substances

[0061] 1) Inoculate the strain Rugamonas sp.A3 on the Gaoshi No. 1 plate by streaking and separating it, and culture it at 27°C for 2-3 days. When the single colonies on the plate begin to turn white to blue, pick the ones that are still white. Inoculate a single colony in the Gaoshi No. 1 liquid medium prepared with distilled water, and culture it at 180 rpm at 27°C for 2 to 3 days, and wait until the bacterial liquid turns blue completely, which is the required fermentation liquid;

[0062] 2) Centrifuge the fermentation broth in step 1) at 12000-14000g for 10-20min;

[0063] 3) Remove the supernatant in step 2), extract the blue bacteria obtained in step 2) with absolute ethanol, and after ultrasonic oscillation for 1 hour, centrifuge at 12000-14000 g for 10-20 minutes;

[0064] 4) Place the supernatant obtained in step 3) under a rotary evaporator at 30° C. and evaporate to dryness ...

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Abstract

The invention discloses an alga inhibiting active substance violacein and a preparation method thereof and relates to a strain of alga inhibiting bacteria and an alga inhibiting active substrate. The strain of the alga inhibiting active compound is Rugamonas sp.A3. The molecular formula of the alga inhibiting active substrate violacein is C18H12O2N6, and the molecular weight is 343.1. The preparation method comprises the following steps: preparing fermentation liquid from Rugamonas sp.A3 obtained by isolation and screening, and centrifuging to obtain bacteria; performing extraction and centrifugation; fetching the supernate and evaporating to dryness; extracting, separating to remove impurities and spinning to dryness to obtain a crude extract; dissolving the crude extract with methanol; loading, eluting and collecting the components with different colors; evaporating to dryness and verifying the alga inhibiting activity, and selecting the components with an alga inhibiting effect; eluting the effective components through a column; collecting different components and performing chromatography; developing and merging similar components; evaporating to dryness and verifying the alga inhibiting activity; selecting the components with alga inhibiting activity; and detecting the purity and detecting the obtained alga inhibiting active components with mass spectrum and 1H-NMR.

Description

technical field [0001] The invention relates to an algae-inhibiting bacterium and an algae-inhibiting active substance, in particular to a violacein with a strong algae-inhibiting active substance and a preparation method thereof. Background technique [0002] The marine ecological abnormal phenomenon that the planktonic microalgae, protozoa or bacteria in the ocean multiply or gather under suitable environmental conditions, causing the deterioration of water quality and discoloration of seawater, is called red tide, also known as harmful algal bloom. [1] . Heterosigma akashiwo, a species of Raphidophytes, is a typical harmful red tide algae that causes fish death. Red tides of Heterosigma akashiwo have occurred in many countries in the northern and southern hemispheres of the world, causing a large Economic losses [2-3] . [0003] The current red tide control methods mainly include: physical methods, chemical methods and biological methods [4] . Physical methods mainly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C07D401/14C12P17/16A01N43/38A01P13/00C12R1/01
CPCA01N43/38C07D401/14C12N1/20C12P17/165C12Q1/04C12N1/205C12R2001/01
Inventor 郑天凌杨旭俊蔡冠竟郑伟
Owner XIAMEN UNIV
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