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Method for preparing high-purity phycoerythrin through hydrophobic chromatography

A phycoerythrin and hydrophobic chromatography technology is applied in chemical instruments and methods, depsipeptides, peptides, etc., to achieve the effects of low cost, simple and easy operation method and high recovery rate

Inactive Publication Date: 2016-11-16
YANTAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, people mainly use sulfuric acid to prepare phycoerythrin by combining fractional precipitation, hydroxyapatite column chromatography, and gel filtration chromatography. The existing preparation process has defects such as complicated operation, high production cost, and unstable product quality, which seriously restrict the application of phycoerythrin.

Method used

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  • Method for preparing high-purity phycoerythrin through hydrophobic chromatography
  • Method for preparing high-purity phycoerythrin through hydrophobic chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Centrifuge to collect 10 g of freshly cultivated Porphyridium algae, and fully mix the algae mud with phosphate buffer (pH7.0, 0.02mol / L) in a ratio of 1:6 to obtain a cell suspension;

[0018] (2) Place the cell suspension in a -20°C environment to freeze completely, take it out and place it at 4°C to thaw, and then place it in a -20°C environment after it is completely thawed, and repeat freezing and thawing 5 times to break the wall liquid; freeze and centrifuge the broken wall liquid, and collect the supernatant;

[0019] (3) At room temperature, the supernatant was ultrafiltered with a membrane bag of 60kD through a molecular cut-off, ultrafiltered 5 times, and the filtrate was collected to obtain a crude phycoerythrin extract;

[0020] (4) Apply the crude phycoerythrin extract obtained in step (3) to a hydrophobic chromatography column, first wash it with 6 times phosphate buffer (pH7.0, 0.02mol / L), and then elute it with distilled water, collect A 545 / A...

Embodiment 2

[0023] (1) Centrifuge to collect 30 g of freshly cultivated Porphyridium algae, mix the algae mud with phosphate buffer (pH 6.8, 0.02mol / L) in a ratio of 1:5 to obtain a cell suspension;

[0024] (2) Place the cell suspension in a -20°C environment to freeze completely, take it out and place it at a temperature of 4°C to thaw, and then place it in a -20°C environment after it is completely thawed, and repeat freezing and thawing 6 times to obtain broken walls liquid; freeze and centrifuge the broken wall liquid, and collect the supernatant;

[0025] (3) At room temperature, the supernatant was ultrafiltered by a membrane bag with a molecular cut-off of 50kD, ultrafiltered 6 times, and the filtrate was collected to obtain a crude phycoerythrin extract;

[0026] (4) Apply the crude phycoerythrin extract obtained in step (3) to a hydrophobic chromatography column, first wash the impurity with 6 times phosphate buffer (pH 6.8, 0.02mol / L), then elute with distilled water, and colle...

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Abstract

The invention provides a method for preparing high-purity phycoerythrin through hydrophobic chromatography. The method comprises the following steps of using fresh porphyridium cruentum as a raw material, uniformly mixing porphyridium cruentum paste with a phosphate buffer solution according to a certain proportion, and performing repeated freezing and thawing for 5-7 times so as to obtain a wall-breaking solution; performing refrigerated centrifugation on the wall-breaking solution, taking a supernatant, and performing ultrafiltration on the supernatant with a membrane bag of which the molecule interception is 50-60kD for 4-6 times so as to obtain a coarse phycoerythrin extract; and then loading samples of the coarse phycoerythrin extract to a hydrophobic chromatography column (Phenyl Sepharose Fast Flow) for gradient elution, collecting flow-out components of which A<545> / A<280> is greater than 4.0, and performing freeze drying so as to obtain purified phycoerythrin. The preparation method disclosed by the invention is good in purification effects, high in protein recovery rate and simple and convenient in technology, and has a good popularization value and good application prospects.

Description

technical field [0001] The invention belongs to the technical field of preparation of marine biological active substances, and in particular relates to a method for preparing phycoerythrin by hydrophobic chromatography. Background technique [0002] Phycoerythrin (PE) is an important light-harvesting pigment protein of a variety of algae, and it forms special particles (phycobili Body), plays the role of absorbing and transmitting light energy. Phycoerythrin is composed of apoprotein and chromophore, and its apoprotein is a kind of oligomeric protein, the basic units are α and β subunits, and some phycoerythrins also have a small amount of γ subunit. The ring-opened tetrapyrrole phycoerythrin is used as the chromophore. Due to the existence of the chromophore, phycoerythrin has strong light absorption at 480-570nm in the visible light region, and has strong fluorescence. Phycoerythrin has a wide range of uses. It can be used as a natural pigment in food, cosmetics and othe...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/20
CPCC07K14/795
Inventor 唐志红赵吉路鞠宝
Owner YANTAI UNIV
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