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Method used for preparing high-purity phycoerythrin via chitosan adsorption

A phycoerythrin, high-purity technology, applied in the preparation method of peptides, chemical instruments and methods, algae/bryopeptides, etc., can solve the problems of complex operation, high production cost, dependence, etc., and achieve simple and easy operation, The effect of low cost and high recovery rate

Inactive Publication Date: 2017-03-15
YANTAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are relatively few domestic studies on it. At present, similar products used in China are all dependent on imports, and their high cost has become a limiting factor for the large-scale use of phycoerythrin in industry and medicine. Therefore, it is very important to develop this type of new marine product.
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 complex operations, high production costs, and unstable product quality. , and it is difficult to achieve large-scale industrial production

Method used

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  • Method used for preparing high-purity phycoerythrin via chitosan adsorption
  • Method used for preparing high-purity phycoerythrin via chitosan adsorption

Examples

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Effect test

Embodiment 1

[0021] (1) Centrifuge to collect 10 g of freshly cultivated Porphyridium algae, mix the algae mud with phosphate buffer in a ratio of 1:5 to obtain a cell suspension;

[0022] (2) The cell suspension obtained in step (1) was repeatedly frozen and thawed 5 times to obtain a wall-breaking liquid; the wall-breaking liquid was refrigerated and centrifuged at 10000r / min for 10min, and the supernatant was collected;

[0023] (3) Add chitosan solution dropwise to the supernatant obtained in step (2) until the mass concentration of chitosan is 0.5%, fully mix, freeze and centrifuge at 10000r / min for 10min, then take the precipitate; add Phosphate buffer solution containing 0.1mol / L sodium chloride, mix thoroughly, and centrifuge at 10,000r / min for 10min to take the precipitate; then add phosphate buffer solution containing 0.3mol / L sodium chloride to the precipitate, mix thoroughly After uniformity, freeze and centrifuge at 10000r / min for 10min, take the supernatant; take a small amou...

Embodiment 2

[0028] (1) Centrifuge to collect 30 g of freshly cultivated Porphyridium algae, mix the algae mud and phosphate buffer in a ratio of 1:8 to obtain a cell suspension;

[0029] (2) The cell suspension obtained in step (1) was repeatedly frozen and thawed 6 times to obtain a wall-breaking liquid; the wall-breaking liquid was refrigerated and centrifuged at 10000r / min for 10min, and the supernatant was collected;

[0030] (3) Chitosan solution is added dropwise to the supernatant obtained in step (2) until the chitosan mass concentration is 0.3%, fully mixed, refrigerated and centrifuged at 10000r / min for 10min, and the precipitate is taken; 0.1mol / L sodium chloride phosphate buffer solution, mix well, then freeze and centrifuge at 10000r / min for 10min, take the precipitate, then add 0.3mol / L sodium chloride phosphate buffer solution to the precipitate, mix well Afterwards, refrigerate and centrifuge at 10000r / min for 10min, and take the supernatant;

[0031] (4) At room temperat...

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Abstract

The invention provides a method used for preparing high-purity phycoerythrin from porphyridium cruentum. The method is simple in operation, and low in cost, and is convenient for large-scale industrialized production. Compared with the prior art, the method used for preparing high-purity phycoerythrin via chitosan adsorption comprises following advantages: a step of column chromatography is avoided; operation is convenient; cost is low; recovery rate is high; purity is high; and verification using laboratory amplify production equipment is carried out. The purity of phycoerythrin obtained via the method A<545> / A<280> is higher than 4.0; analytical grade requirements are satisfied; and foundation is provided for applications of phycoerythrin in the fields such as biomedical analysis and immunochemistry.

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 Porphyrophycoerythrin by adopting a chitosan adsorption method. Background technique [0002] Porphyridium cruentum belongs to the unicellular algae of Rhodophyta, which can produce a variety of biologically active substances such as: phycobiliproteins, polyunsaturated fatty acids and exopolysaccharides. The phycoerythrin contained in it is a kind of light-harvesting pigment protein. It is composed of apoprotein and chromophore. The apoprotein is a kind of oligomeric protein, and the ring-opened tetrapyrrole phycoerythrin is used as part of the chromophore. Due to the existence of chromophore, phycoerythrin is visible in the visible light region at 480 It has strong light absorption at ~570nm and has strong fluorescence. Phycoerythrin has a wide range of uses. It can be used as a natural pigment in fo...

Claims

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Application Information

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IPC IPC(8): C07K14/405C07K1/36C07K1/34C07K1/30
CPCC07K14/405
Inventor 唐志红徐静孙利芹任育红王晓倩丁珂季昱含
Owner YANTAI UNIV
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