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Separating and purifying process of isochrysis galbana exopolysaccharide

A technology for the separation and purification of dinoflagellate cells, applied in the field of marine biochemical engineering, can solve the problems of polysaccharide separation and purification, structure identification and physiological activity that have not been reported, and achieve good operability, stable reproducibility, simple and reasonable process Effect

Inactive Publication Date: 2012-12-26
HUAIHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with the research on other microalgal polysaccharides, the research on Isochrysis globosa polysaccharides is still in its infancy, and the separation and purification, structural identification and physiological activities of its polysaccharides have not been reported yet.

Method used

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  • Separating and purifying process of isochrysis galbana exopolysaccharide
  • Separating and purifying process of isochrysis galbana exopolysaccharide
  • Separating and purifying process of isochrysis galbana exopolysaccharide

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1, with reference to figure 1 , a separation and purification process of Isochrysis globosa exopolysaccharide, the steps are as follows:

[0023] (1) Select the culture of Isochrysis globosa that has been cultured to the exponential growth stage, centrifuge at 5000 rpm / min for 15 min, take the supernatant, filter it through a 0.22 μm microporous membrane, concentrate under reduced pressure at 45°C; freeze-dry After that, a white powdery extracellular substance was obtained;

[0024] (2) Take extracellular substances and add them to distilled water, the mass ratio of distilled water to extracellular substances is 15:1. After mixing evenly, adjust the pH to 9.0 with 0.5 M NaOH, and extract in a water bath at 70°C for 240 min; the extraction is completed Afterwards, centrifuge at 5000 rpm / min for 5 min, take the supernatant and add trichloroacetic acid with a mass concentration of 3%, let stand at 4 °C for 4 h, centrifuge at 5000 rpm / min for 5 min, and discard...

Embodiment 2

[0028] Embodiment 2, with reference to figure 1 , a separation and purification process of Isochrysis globosa exopolysaccharide, the steps are as follows:

[0029] (1) Select the culture of Isochrysis globosa that has been cultured to the exponential growth stage, centrifuge at 5000 rpm / min for 15 min, take the supernatant, filter it through a 0.22 μm microporous membrane, concentrate under reduced pressure at 45°C; freeze-dry After that, a white powdery extracellular substance was obtained;

[0030] (2) Take extracellular substances and add them to distilled water, the mass ratio of distilled water to extracellular substances is 15:1. After mixing evenly, adjust the pH to 9.0 with 0.5 M NaOH, and extract in a water bath at 70°C for 240 min; the extraction is completed Afterwards, centrifuge at 5000 rpm / min for 5 min, take the supernatant and add trichloroacetic acid with a mass concentration of 3%, let stand at 4 °C for 4 h, centrifuge at 5000 rpm / min for 5 min, and discard...

Embodiment 3

[0034] Embodiment 3, the separation and purification process experiment 1 of Isochrysis globosa exopolysaccharide, the steps are as follows:

[0035] Weigh 0.5 g of the extracellular crude polysaccharide of Isochrysis globosa produced in step (2) of Example 1, dissolve it in 100 mL of distilled water, load it on DEAE-52 ion exchange column chromatography, and first elute with distilled water, Collect 3 mL of the tube at a flow rate of 1.5 mL / min, and detect polysaccharides by the sulfuric acid-anthrone method until no polysaccharide components are detected. The polysaccharide fractions were collected together and concentrated under reduced pressure to 10 mL to obtain the neutral polysaccharide fraction (ECPSⅠ). Subsequently, the elution was continued with 1.0 mol / L NaOH, and 3 mL was collected in each tube at a flow rate of 1.5 mL / min. Polysaccharides were detected by the sulfuric acid-anthrone method until no polysaccharide components were detected. The polysaccharide fracti...

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Abstract

The invention provides a separating and purifying process of isochrysis galbana exopolysaccharide; and the process comprises the following steps of: preparing crude isochrysis galbana exopolysaccharide by using isochrysis galbana culturing solution as a raw material; separating the crude isochrysis galbana exopolysaccharide ECPS by ion exchange chromatography, so as to obtain neutral polysaccharide and acidic polysaccharide components; loading the neutral polysaccharide and acidic polysaccharide components on a gel column for chromatography, so as to obtain eight components, wherein except for an ECPSII-A component, the quantities of the rest polysaccharide components are small; purifying the ECPSII-A component by using the gel column chromatography, so as to obtain a single polysaccharide component; and depositing, freezing and drying the single polysaccharide component by ethanol so as to prepare pure exopolysaccharide ECPSIII. According to the invention, the separating and purifying process of the isochrysis galbana exopolysaccharide is established; and the process is simple and reasonable and has the advantages of stable reproducibility and good operability. A good experiment basis is laid for follow-up study on biological activity of the polysaccharide; moreover, the microalgae polysaccharide can be applied to the medicine and medical field like other polysaccharide substances.

Description

technical field [0001] The invention belongs to the field of marine biochemical engineering, and in particular relates to a process for separating and purifying the exopolysaccharide of Isoflagellates. Background technique [0002] Isochrysis globosa ( Isochrysis galbana ) are rich in polyunsaturated fatty acids, proteins and polysaccharides, rich in nutrients, and are commonly used bait microalgae in aquaculture. Previous studies have mainly focused on polyunsaturated fatty acids. In recent years, with the in-depth study of microalgal polysaccharides, the research on Isochrysis globosa polysaccharides has been increasing. However, compared with other microalgae polysaccharides, the research on Isochrysis globosa polysaccharides is still in its infancy, and the separation, purification, structure identification and physiological activity of its polysaccharides have not been reported yet. Among them, there are fewer studies on its exopolysaccharides. [0003] During the g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00
Inventor 孙颖颖
Owner HUAIHAI INST OF TECH
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