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High-efficiency algae-inhibiting active compound deinoxanthin and its preparation method and application

A technology of algae-inhibiting activity and compounds, which is applied in the field of high-efficiency algae-inhibiting active compound Deinoxanthin and its preparation, and can solve the problem of insufficient algae-inhibiting substances to meet harmful red tides

Inactive Publication Date: 2017-02-08
XIAMEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research on the relationship between bacteria and algae in China lags behind that of foreign countries, and the research on the extraction, purification and identification of algae-inhibiting substances is still in its infancy. The isolated and identified algae-inhibiting substances are far from enough to meet the prevention and control of harmful red tides

Method used

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  • High-efficiency algae-inhibiting active compound deinoxanthin and its preparation method and application
  • High-efficiency algae-inhibiting active compound deinoxanthin and its preparation method and application
  • High-efficiency algae-inhibiting active compound deinoxanthin and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: Separation and screening of anti-algae bacterial strains

[0068] 1) Take a lake water sample from the Xiang’an Campus of Xiamen University in Fujian, dissolve it in 90 mL of autoclaved LB medium (10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride, pH 7.2, 1 L of distilled water), and place it on a shaker at 150-200 rpm Shake for 20-30 minutes to disperse the sample evenly;

[0069] 2) 10-fold dilution method, spread evenly dispersed samples on LB solid plate, and culture at 28-37°C for 3-5 days;

[0070] 3) Pick different types of single colonies and streak them on LB solid plates, culture them at 28-30°C for 3-5 days, verify whether they are pure cultures, and repeat this step until pure cultures are obtained ( figure 1 ,2);

[0071] 4) Inoculate a single colony of the isolated pure culture into 4 mL of LB liquid medium, place it on a shaker at 28-37°C, and culture with shaking at 180-250 rpm for 3-5 days;

[0072] 5) Add 1mL of fermentati...

Embodiment 2

[0073] Embodiment 2: Algae inhibiting effect assay method

[0074] 1) Alexandrium tamar 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;

[0075] 2) Quantitatively add the sample to be tested into a 24-well plate;

[0076] 3) Take Alexandrium tamarensis culture solution 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. At the same time, the morphological changes of algal cells were observed.

Embodiment 3

[0077] Embodiment 3: Separation and identification of anti-algae active compound

[0078] 1) Inoculate Deinococcus xianganensis Y35 on a plate, streak and separate it, and culture it at 28-37°C for 2-3 days; pick a single colony on the plate and inoculate it in LB liquid medium prepared with distilled water , at 28-37°C, 180-250rpm for 2-3 days to obtain the fermentation broth;

[0079] 2) Centrifuge the fermentation broth in step 1) at 12000-14000g for 10-20min;

[0080] 3) Remove the supernatant in step 2), extract the red bacteria obtained in step 2) with absolute ethanol, and after ultrasonic oscillation for 2-4 hours, centrifuge at 12000-14000 g for 10-20 min;

[0081] 4) Put the supernatant obtained in step 3) into a rotary evaporator at 30°C to dry in vacuo, add 200-400 mL of ethyl acetate to extract overnight, concentrate under reduced pressure, and dry to obtain crude extract A;

[0082] 5) Dissolve the crude extract A obtained in step 4) in methanol, then apply it ...

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Abstract

The invention discloses a high-efficiency algal-inhibition active compound Deinoxanthin and a preparation method and application thereof and relates to a stain of algal-inhibition bacterium and an algal-inhibition active compound. The compound is C40H54O3. Y35 is inoculated to a panel to be subjected to streaking; after Y35 is cultured, a single colony is selected to be cultured again to obtain fermentation liquor; centrifugation is carried and supernatant is removed; the obtained bacterium is extracted and centrifuged; the supernatant is evaporated to dryness under the vacuum condition and is extracted, decompressed, concentrated and dried to obtain crude extract; then the crude extract is dissolved in methyl alcohol; a sample is loaded to a sephadex column; elution, chromatography and color development are carried out; similar components are combined to obtain combined eluent; then after being evaporated to dryness under the vacuum condition, the combined eluent is dissolved into DMSO (dimethyl sulfoxide) to be subjected to algal-inhibition activity verification; components with algal-inhibition activity are selected and then are dissolved in ethyl acetate; the obtained sample is loaded to a silica gel column; elution is carried out; eluent is collected interruptedly; chromatography is carried out; after color development, similar components are combined to obtain the combined eluent; the obtained product is evaporated to dryness under the vacuum condition and then is dissolved in DMSO to verify algal-inhibition activity; and the components with algal-inhibition activity are selected and are analyzed by high performance liquid chromatography to obtain the product.

Description

technical field [0001] The invention relates to an algae-inhibiting bacterium and an algae-inhibiting active compound, in particular to a high-efficiency algae-inhibiting active compound Deinoxanthin and its preparation method and application. Background technique [0002] The overgrowth of marine algae will form red tides, and the outbreak of red tides often causes changes in the marine environment and destroys the original ecological balance [1]. In particular, the red tide caused by toxic algae, the algae toxins secreted by it caused the death of marine life, which in turn caused huge economic problems and health crises [2,3]. Alexandrium tamaris is a kind of toxic dinoflagellate distributed globally, and its toxins are transmitted through the food chain to form paralytic shellfish poisoning, which is extremely harmful to the water environment and human health, and often causes red tides in Xiamen waters[4 ,5], is a toxic algae that urgently needs to be treated and contr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P23/00C07C49/743C07C45/78A01N35/06A01P13/00C12R1/01
Inventor 郑天凌李祎朱红张化俊郑伟
Owner XIAMEN UNIV
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