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Separation and purification method for Dunaliella salina polysaccharide

A technique for the separation and purification of polysaccharides from Dunaliella salina, applied in the biotechnology of salina salina and its products, and in the field of chromatographic separation, can solve the problems of lack of systematic and in-depth research on the separation and purification of polysaccharides, and the lack of research and utilization. The method is simple and practical, and is conducive to popularization , low cost effect

Inactive Publication Date: 2009-06-03
JIANGNAN UNIV
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Problems solved by technology

However, at present, large-scale cultivation of Dunaliella salina is mainly used to produce β-carotene, and the research and utilization of polysaccharides as high as 10%-40% are less, and there is a lack of systematic and in-depth research on the separation and purification of its polysaccharides. The reported salina polysaccharide products are all glycoproteins or heteropolysaccharides mainly composed of glucose. The relevant application of ion exchange chromatography-phosphate buffer ionic strength gradient elution combined with gel filtration chromatography-NaAc aqueous solution gradient elution separation The method for preparing nucleic acid polysaccharides and sulfated heteropolysaccharides mainly containing galactose and containing relatively high levels of arabinose and xylose from Salina has not been reported.

Method used

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  • Separation and purification method for Dunaliella salina polysaccharide
  • Separation and purification method for Dunaliella salina polysaccharide
  • Separation and purification method for Dunaliella salina polysaccharide

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Take 200g of Dunaliella salina powder, heat and extract with 2000mL of pH 8.5 NaOH alkaline aqueous solution in a water bath at 90°C for 3.0 hours, extract twice, and combine to obtain the extract; adjust the pH of the extract to 4.0 with 2mol / L HCl, and centrifuge (4000r / min) 20min, after concentrating the supernatant, use 3.8 times of 95% ethanol alcohol precipitation, low temperature (4 ℃) for 8 hours, centrifugal, suction filtration, take the filter cake and vacuum dry to obtain the crude extract; The free protein was removed by Sevag method, followed by dialysis and concentration to obtain the crude polysaccharide concentrate which was preliminarily purified. After freeze-drying the crude polysaccharide concentrate, the polysaccharide content was determined by the phenol-sulfuric acid method to be 75.5%, and the yield was 9.0%.

[0018] Load 50 mL of the crude polysaccharide concentrate that was initially purified on a DEAE-Sepharose Fast folw column (3.5 cm × 30 c...

Embodiment 2

[0021] Take 500g of Dunaliella salina powder, heat and extract with 7500mL of pH9.0 alkaline aqueous solution at 80°C in a water bath for 4 hours, extract twice, and combine to obtain the extract; adjust the pH of the extract to 3.5 with 2mol / L HCl, and centrifuge (4000r / min) 20min, the obtained supernatant was concentrated, and the polysaccharide was precipitated with 3.8 times the volume of 95% ethanol, left standing overnight at low temperature (4°C), centrifuged, suction filtered, and the filter cake was vacuum-dried to obtain the crude extract; further use Sevag The free protein was removed by the method, and the crude polysaccharide concentrate was obtained by dialysis and concentration. After freeze-drying the crude polysaccharide concentrate, the polysaccharide content was determined by the phenol-sulfuric acid method to be 78.2%, and the yield was 8.5%.

[0022] Load 50 mL of the initially purified crude polysaccharide aqueous solution (10 mg / mL) on a DEAE-Sepharose ...

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Abstract

The invention relates to a separation and purification method for Dunaliella salina polysaccharide, which belongs to the technical field of salina biotechnology and the product and chromatographic fractionation thereof. The method comprises the following steps: using beta-carotene extracted Dunaliella salina powder as the raw material and performing hot water extraction on alkaline condition, removing most part of protein by the isoelectric point method, separating out polysaccharide by the ethyl alcohol precipitation method, removing the residual protein by the Sevag method, performing separation and purification by the ion-exchange chromatography-phosphate buffer ionic strength gradient elution method in combination with gel filtration chromatogram-NaAc aqueous solution gradient elution method, collecting separately, freeze drying after appropriate concentration, and finally obtaining five homogeneous components of Dunaliella salina polysaccharide, including sulphatized heteropolysaccharide containing galactose as the main part and high-content arabinose and xylose, and nucleic acid polysaccharide. The method has the advantages of simplicity, practicability and low cost, thereby being favorable for promotion, development and application.

Description

technical field [0001] The invention discloses a method for separating and purifying polysaccharides from Dunaliella salina, relating to a method for separating and purifying sulfated heteropolysaccharides and nucleic acid polysaccharides from Dunaliella salina, and belongs to the technical field of salina biotechnology, its products, and chromatographic separation. Background technique [0002] Duanaliella salina is a single-celled green algae plankton without cell walls. It is not only an important type of marine microalgae, but also can be cultured in inland salt lakes. There are abundant primary and secondary metabolites with unique composition and various physiological activities in Dunaliella salina—in addition to high content of natural β-carotene, it also contains rich polysaccharides, proteins, unsaturated fatty acids, etc. A variety of biologically active substances and minerals needed by the human body. These physiologically active substances have good applicatio...

Claims

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Application Information

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IPC IPC(8): C08B37/00C08B37/02
Inventor 戴军王旻陈尚卫朱松汤坚尹鸿萍
Owner JIANGNAN UNIV
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