Method for extracting blue-green algae extracellular polymeric substance in grading manner

A technology for extraction of extracellular polymers and graded extraction, which is applied in the field of extraction of extracellular polymers from cyanobacteria, can solve the problems of poor comparability of results and large differences in extraction operating conditions, etc., to achieve increased yield, ensure cell integrity, and high extraction efficiency Effect

Inactive Publication Date: 2014-12-31
NANJING INST OF GEOGRAPHY & LIMNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although there are many studies on extracellular polymers, the results obtained by different researchers are not comparable, partly because of the large differences in the extraction conditions they adopted, and another important reason is that they all use extracellular polymers Taken as a whole, no fractional extraction of extracellular polymers was performed
However, there are no relevant reports on fractional extraction of cyanobacterial extracellular polymeric substances

Method used

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  • Method for extracting blue-green algae extracellular polymeric substance in grading manner
  • Method for extracting blue-green algae extracellular polymeric substance in grading manner
  • Method for extracting blue-green algae extracellular polymeric substance in grading manner

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Take a sample of cyanobacteria water body in Taihu Lake in summer. The biomass of cyanobacteria is 0.8g / L. After the cyanobacteria water sample is collected, it is placed in a refrigerator at 4°C, and the extracellular polymer is fractionated and extracted within 12 hours. The specific operation steps are as follows:

[0031] (1): Extraction of soluble and loosely bound extracellular polymers from cyanobacteria

[0032] Take a certain amount (50ml) of cyanobacteria water samples and centrifuge at 2500g centrifugal force for 15min at 4°C. The supernatant collected is SL-EPS; Centrifuge at 4500g for 15 minutes, and the collected supernatant is LB-EPS; add 0.05% NaCl buffer solution to the original volume of the solid sample, and proceed to the next step.

[0033] (2): Extraction of tight-binding extracellular polymers from cyanobacteria

[0034] Use 40% NaOH solution to adjust the pH of the algal cells to be extracted to 9.0, equilibrate for 10 minutes, and then place t...

Embodiment 2

[0041] Take the cyanobacteria water samples in Taihu Lake in autumn. The cyanobacteria biomass is 0.5g / L. After the cyanobacteria water samples are collected, they are placed in a 4°C refrigerator, and the extracellular polymers are fractionated and extracted within 12 hours. The specific operation steps are as follows:

[0042] (1): Extraction of soluble and loosely bound extracellular polymers from cyanobacteria

[0043] Take a certain amount (50ml) of cyanobacteria water samples and centrifuge at 2000g centrifugal force for 15min at 4°C. The collected supernatant is SL-EPS; add 0.07% NaCl buffer solution to the original volume of solid samples, Centrifuge at a centrifugal force of 5000g for 15 minutes, and the collected supernatant is LB-EPS; add 0.07% NaCl buffer solution to the original volume of the solid sample, and proceed to the next step.

[0044] (2): Extraction of tight-binding extracellular polymers from cyanobacteria

[0045] Use 35% KaOH solution to adjust the...

Embodiment 3

[0050] Microcystis aeruginosa was cultivated indoors to obtain a culture solution of Microcystis aeruginosa, and the algae biomass was determined to be 0.15 g / L. Samples were subjected to the following steps:

[0051] Take a certain amount (50ml) of the culture solution sample and centrifuge at 2500g centrifugal force for 15min at 4°C, and the collected supernatant is SL-EPS; add 0.08% NaCl buffer solution to the original volume of the solid sample, Centrifuge at 4500g for 15 minutes, and the collected supernatant is LB-EPS; add 0.08% NaCl buffer solution to the original volume of the solid sample, and proceed to the next step.

[0052] (2): Extraction of tight-binding extracellular polymers from cyanobacteria

[0053] Use 40% KaOH solution to adjust the algae cells to be extracted to pH 8.0, balance for 10 minutes, and then place the algae liquid to be extracted at 55 o C constant temperature hot water reaction for 30min. Take it out and cool it immediately, and centrifuge...

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Abstract

The invention relates to a method for extracting a blue-green algae extracellular polymeric substance in a grading manner. The method comprises the following steps of: firstly, carrying out middle / low-speed eccentric shearing to obtain a soluble and loosely combined type extracellular polymeric substance; and secondly, extracting a tightly combined type extracellular polymeric substance in a mode of subalkaline hot bath with the combination of high-speed eccentric shearing. The physical and chemical analysis result shows that the soluble and loosely combined type extracellular polymeric substance contains a small amount of organisms, and the tightly combined type extracellular polymeric substance contain a great amount of organisms; and three-dimensional fluorescence and infrared spectroscopy result shows that the soluble extracellular polymeric substance is only composed of polysaccharide, protein substances and humus substances, and the loosely combined and tightly combined extracellular polymeric substances only consist of polysaccharide and protein substances. The invention discloses an operation method for grading extraction of the blue-green algae extracellular polymeric substance. The method has the advantages that the operation is simple, the cyanobacterium is free of damage, the cyanobacterium is analyzed in different grades, and the like; and the method has high guiding significance in industrial application of the blue-green algae extracellular polymeric substance.

Description

technical field [0001] The invention relates to a method for extracting cyanobacterial extracellular polymers, in particular, the method can realize the graded extraction operation of extracellular polymers, thereby obtaining cyanobacterial extracellular polymers with different organic components and contents, and belongs to the technical field of bioengineering . Background technique [0002] In recent years, with the intensification of human activities and eutrophication, lake cyanobacteria blooms frequently broke out, causing adverse effects on human health and the ecological environment. Cyanobacterial extracellular polymer is a general term for a series of macromolecular organic substances produced during the growth and reproduction of cyanobacteria cells, mainly derived from cell secretion, cell lysis, cell adsorption and foreign substances. Cyanobacterial extracellular polymers form cyanobacterial colonies by adhering to individual cyanobacteria, thereby promoting th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14C08B37/00
Inventor 江和龙徐华成
Owner NANJING INST OF GEOGRAPHY & LIMNOLOGY
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