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Method for extracting streptomyces alga dissolution active substances

An active substance and extraction method technology, applied in the field of eutrophic water bioremediation, can solve the problems of increased difficulty in separation and purification of algae-dissolving active substances, low concentration of algae-dissolving substances, unstable properties, etc., and achieves good algae-dissolving effect, The effect of low production cost and high safety

Inactive Publication Date: 2018-05-18
无锡市拜沃特环保科技有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the algae-dissolving active substances secreted by microorganisms are highly polar substances, and most of the algae-dissolving substances have low concentration and unstable properties, and are very likely to be lost in the process of concentration and extraction; in addition, the process of microbial cultivation and reproduction usually contains a large number of other compounds , and the properties of algicidal substances secreted by different algicidal bacteria are different. The above factors make the separation and purification of algicidal active substances more difficult.

Method used

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  • Method for extracting streptomyces alga dissolution active substances
  • Method for extracting streptomyces alga dissolution active substances
  • Method for extracting streptomyces alga dissolution active substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: experimental material and method

[0031] Microcystis aeruginosa ( Microcystis aeruginosa FACHB-905), Astringentia ( Aphanizomenon flos-aquae FACHB-1039), Anabaena ( Anabaena flos-aquae FACHB-1092) was provided by the Algae Collection Center of the Institute of Hydrobiology, Chinese Academy of Sciences. After activation, the algae were inoculated in BG11 medium and cultivated to the logarithmic phase for later use.

[0032] Algae-dissolving active components M1, M2, M3, M4 and M5 were dissolved with methylene blue and adjusted to 50 mL respectively, and inoculated at 5% in 10 mL algae liquid with an initial chlorophyll a concentration of 0.1521 mg / L, and co-cultured for 4 days Then measure the content of chlorophyll a and calculate the algae inhibition rate.

[0033] The algal species and algae-dissolving experiment were cultivated under the conditions of 25 ℃, 2500 lx and a light-dark ratio of 14 h:10 h.

[0034] Chlorophyll a content was determ...

Embodiment 2

[0038] Embodiment 2: Streptomyces algicidal active substance extraction method

[0039] A method for extracting streptomyces algae-dissolving active substances, comprising the steps of:

[0040] a. Streptomyces was inoculated in 1 L of liquid Gaoshi No. 1 medium, and fermented at 28 °C and 100 rpm for 4 days to obtain a fermentation broth;

[0041] b. Centrifuge the fermented liquid obtained in step a at 4°C and 10,000 rpm for 15 min, and filter the supernatant through glass fiber filter paper with a pore size of 0.45 μm to obtain the filtrate;

[0042] c. The filtrate obtained in step b was repeatedly extracted three times with an equal volume of absolute ethanol, and after centrifugation, the organic phase was concentrated to dryness with a rotary evaporator to obtain a crude substance A45 mg;

[0043] d. Dissolve the crude substance A obtained in step c in 2 ml of methanol, load the sample for silica gel column chromatography, and use chloroform and methanol at a volume ra...

Embodiment 3

[0048] Example 3: Inhibition effect of algae-dissolving active substances on Microcystis aeruginosa

[0049]Dissolve the algae-dissolving active substance H of the present invention with methylene blue and set the volume to 50 mL, and inoculate it in 100 mL of chlorophyll a concentration of 0.1521 mg / L at the volume ratio of 0, 1%, 3%, 5% and 10% respectively. In Microcystis aeruginosa algae liquid, after co-cultivation for 4 days at 25 ℃, 2500 lx and light-dark ratio of 14 h:10 h, the content of chlorophyll a was measured and the algae inhibition rate was calculated. The results showed that the algae-dissolving active substance H had a significant inhibitory effect on Microcystis aeruginosa, and the algae inhibition rates were 91.3%, 93.9%, 94.2% and 97.6% when the addition amount was 1%, 3%, 5% and 10%, respectively. Adding 1% can effectively inhibit the growth of Microcystis aeruginosa.

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Abstract

The invention provides a method for extracting streptomyces alga dissolution active substances, and belongs to the field of eutrophic water body bioremediation. An isometric absolute ethyl alcohol isused for repeatedly extracting streptomyces fermentation liquor filtrate, after centrifugalization is completed, a rotary evaporator is used for concentrating an organic phase to be dry, crude substances are obtained, then methyl alcohol is used for dissolving the crude substances, and silica gel column chromatography separation is carried out; the components with efficient alga dissolution activity are subjected to high-performance liquid chromatographic separation, and the alga dissolution active substances are obtained. Adopted streptomyces comes from an eutrophic water body, the alga dissolution active substances secreted by the streptomyces are secondary metabolites, high safety is achieved, and no secondary pollution is caused to the water body. The alga dissolution active substancescan be used for treatment of the eutrophic water body caused by typical harmful algae such as microcystis, anabaena, aphanizomenon and oscillatoria. The extraction method is simple in process, high in operability, low in production cost, good in alga dissolution effect and suitable for large-scale production.

Description

technical field [0001] The invention belongs to the field of bioremediation of eutrophic water bodies, in particular to a method for extracting streptomyces algae-dissolving active substances, and at the same time relates to the application of streptomyces algae-dissolving active substances. Background technique [0002] Eutrophication not only causes serious harm to the water environment, aquaculture and marine fisheries, but also affects human health and even life. In the case of unsatisfactory control by physical and chemical methods, it is imminent to seek safe and effective ways to prevent and control algal blooms. Alginolytic bacteria are a kind of bacteria that inhibit the growth of algae, kill algae or dissolve algae cells. They have the advantages of high efficiency, low risk of secondary pollution, and high ecological safety. The control of algal biomass can be achieved within a certain period of time, and it is bound to become an important means of biological con...

Claims

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Application Information

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IPC IPC(8): C12P1/06C02F3/34C12R1/465
CPCC12P1/06C02F3/34C12R2001/465C12N1/205
Inventor 孔赟陈亿军邹培邹桐
Owner 无锡市拜沃特环保科技有限公司
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