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Method for extracting microcystis aeruginosa extracellular polymer

A technology of Microcystis aeruginosa cells and extracellular polymers, which is applied in the field of extraction of extracellular polymers of Microcystis aeruginosa

Pending Publication Date: 2018-12-21
温州大学苍南研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no widely recognized and effective method for the extraction and separation of algal EPS. Therefore, researchers who study cyanobacterial EPS and cyanobacterial blooms have been working on exploring an efficient algal EPS extraction method.

Method used

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  • Method for extracting microcystis aeruginosa extracellular polymer
  • Method for extracting microcystis aeruginosa extracellular polymer
  • Method for extracting microcystis aeruginosa extracellular polymer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] 1. Samples of Microcystis aeruginosa: Microcystis aeruginosa FACHB-469 purchased from the Wild Biological Germplasm Bank of the Chinese Academy of Sciences—Freshwater Algae Species Bank, placed in a light incubator and cultured with BG11 medium; the culture temperature is 25°C , the time is set to 12Hr day: 12Hr night, and the light condition is 2000Lux. Take 500ml of algae liquid, filter it, and dry it at 105°C until it reaches a constant weight, and the dry matter of the algae liquid is about 0.6g / L. Before the experiment, the density of the algae liquid algae was measured to be about 7×10 8 per L, the chlorophyll content is about 3800mg / L, and the photosynthetic activity (Fv / Fm) is about 0.4.

[0061] 2. Extraction and separation of EPS from Microcystis aeruginosa

[0062] Specific steps are as follows:

[0063] (1) Take 500ml of Microcystis aeruginosa FACHB-469 algae liquid, centrifuge at 4°C and 2500g centrifugal force for 15 minutes in an ultra-low temperature ...

Embodiment 2

[0067] In this example, different methods are used to extract and separate Microcystis aeruginosa EPS. Except that step (3) is different from Example 1, the rest of the steps are exactly the same as Example 1.

[0068] The processing method of step (3) is as follows:

[0069] Treatment 1 (referred to as TB-1): resuspend the residue of the lower layer of step (2) in 0.05% NaCl buffer solution, and sonicate for 2 min under the conditions of 120W and 25kHz; then, place the buffer solution at 4°C to Centrifuge at 15,000 g for 20 minutes, and take the supernatant to obtain a tightly bound extracellular polymer (TB-EPS).

[0070] Treatment 2 (referred to as TB-2): Resuspend the residue of the lower layer in step (2) in 0.05% NaCl buffer solution, heat in a water bath at 60°C for 30min; then, centrifuge the buffer solution at 4°C with a centrifugal force of 15000g After 20 minutes of operation, the supernatant was taken to obtain a tightly bound extracellular polymer (TB-EPS).

[0...

Embodiment 3

[0093] This example explores the effects of changes in pH value, stirring speed and time, and water bath temperature and time on EPS pollution and three-dimensional fluorescence characteristics. Except that step (3) is different from Example 1, the rest of the steps are the same as in Example 1. 1 is exactly the same.

[0094] The processing method of step (3) is as follows:

[0095] Treatment 1: Resuspend the residue of the lower layer of step (2) in 0.05% NaCl buffer, add sodium hydroxide, adjust the pH of the buffer to 8, 9, 10, 11 respectively, at 4°C and 80rpm After stirring for 10 minutes, place it in a water bath at 60°C and heat it for 30 minutes; then, centrifuge the buffer solution at 4°C with a centrifugal force of 15,000 g for 20 minutes, and take the supernatant to obtain a tightly bound extracellular polymer (TB-EPS) .

[0096] Treatment 2: Resuspend the residue of the lower layer of step (2) in 0.05% NaCl buffer solution, add sodium hydroxide, adjust the pH of...

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Abstract

The invention discloses a method for extracting a microcystis aeruginosa extracellular polymer, The method comprises the following steps of taking an algae liquid of the microcystis aeruginosa, and performing centrifuging to obtain a dissolved type extracellular polymer; suspending the lower layer residue in a buffer solution, and performing centrifuging to obtain a loose attached type extracellular polymer; suspending the lower layer residue in an NaCl buffer solution, and adding sodium hydroxide, adjusting the pH value to be 11, and under the conditions of 4 DEG C and 80-120 rpm, performingstirring for 10-20 minutes, and then putting the mixture into a water bath at 40-60 DEG C for heating for 20-40 minutes; and performing centrifuging on the buffer solution to obtain a tightly combinedtype extracellular polymer. According to the method, the extracellular polymer is extracted by adopting a combination mode of "NaOH and heating" on microcystis aeruginosa, and the parameter extraction conditions are strictly controlled, so that a large amount of EPS in the algae be extracted, the algae cells can not be damaged, and pollution of the EPS is not caused, and therefore, the effectiveEPS component is extracted.

Description

technical field [0001] The invention relates to the field of algal blooms in water bodies, in particular to a method for extracting extracellular polymers of Microcystis aeruginosa. Background technique [0002] Extracellular Polymeric Substance (EPS) secreted by microbial flora has been recognized as an important factor for microbial aggregation and biofilm formation. Cyanobacterial blooms are also accompanied by the production of cyanobacterial extracellular polymeric substances (EPS). Cyanobacterial EPS have also been reported to be involved in the aggregation and development of microorganisms. When the external environmental conditions are suitable for the growth of cyanobacteria, the EPS-producing cyanobacteria colonies in the lake will gather together and float to the water surface to form a cyanobacteria bloom. Xu Huacheng studied the role of EPS in the aggregation of Microcystis and the formation of water blooms, and found that EPS extract can reduce the cohesion o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/44
CPCG01N1/28G01N1/44
Inventor 王奇赵敏于恒国葛姝洁邱颖庞文静李军柯强王传花戴传军贝克黄先锋陈琼珍金展
Owner 温州大学苍南研究院
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