Method for fast separating and purifying R-phycoerythrin, R-phycocyanin

A technology of phycoerythrin and phycocyanin, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve problems such as efficient extraction of R-phycoerythrin, and overcome large-scale rapid preparation, time-consuming Low, high yield effect

Inactive Publication Date: 2008-08-13
SHANDONG UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the defects of the prior art that R-phycoerythrin and R-phycocyanin cannot be extracted simultaneously, in large quantities, and efficiently, the present invention provides a method for rapid and large-scale separation and purification of R-phycoerythrin and R-phycocyanin

Method used

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  • Method for fast separating and purifying R-phycoerythrin, R-phycocyanin
  • Method for fast separating and purifying R-phycoerythrin, R-phycocyanin

Examples

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Embodiment 1

[0031] A fast and efficient separation and purification method for R-phycoerythrin and R-phycocyanin, the steps are as follows:

[0032] (1) Extraction of R-phycoerythrin and R-phycocyanin

[0033] The raw material is natural Polytubuleum, which is stored at -20°C.

[0034] Take 10g of cryopreserved Polytubule algae, add 10ml of 20mM acetate buffer (pH5.8), freeze-thaw repeatedly, centrifuge the suspension at 10000rpm at 4°C for 15min, and the supernatant is R-phycoerythrin and R-phycocyanin Crude protein extract.

[0035] (2) Preliminary purification of R-phycoerythrin and R-phycocyanin

[0036] Add solid ammonium sulfate to the crude extract of R-phycoerythrin and R-phycocyanin obtained in the above step (2) to a concentration of 60% (w / v), leave it for 4h, centrifuge at 10000rpm at 4°C for 15min, collect Precipitation part. The precipitate was dissolved in 20 mM acetate buffer (pH 5.6), and then dialyzed against 20 mM acetate buffer (pH 5.6). Collect the dialysate.

...

Embodiment 2

[0043] The rapid separation and purification method of R-phycoerythrin and R-phycocyanin as described in Example 1, the difference is that in step (2), solid ammonium sulfate is directly added to a concentration of 50% (w / v), Leave it for 4 hours, centrifuge at 10,000 rpm at 4°C for 20 minutes, and collect the precipitated part.

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Abstract

A rapid separation method of purified R-phycoerythrin and R-phycocyanin, which pertains to separation and purification of red algae technical field. The invention uses red algae as raw material, extracts R-phycoerythrin and R-phycocyanin by freeze dissolving and intensified swelling with low ions, carries out primary purification after precipitation with ammonia sulfate, then absorb and enrich R-phycoerythrin and R-phycocyanin by anion exchange chromatography, and further elutes highly purified R-phycoerythrin and R-phycocyanin from anion exchange chromatography step by step. The method eliminates conventional complex separation and purification step which uses multiple chromatography combining molecular sieve chromatography with hydroxyapatite chromatography, solves the problem of extensive and rapid preparation. The method is easy in operation, time and energy saving, with little requirements to apparatus, and high in yield.

Description

technical field [0001] The invention relates to a method for rapid separation and purification of R-phycoerythrin and R-phycocyanin, belonging to the technical field of separation and purification of red algae. Background technique [0002] Phycobiliproteins are a type of water-soluble light-harvesting pigment-protein complexes that exist in cyanobacteria, red algae, cryptophytes and a few dinoflagellates. They play a role in efficiently capturing and transmitting light energy in photosynthesis. Phycocyanin (APC), phycocyanin (PC), phycoerythrocyanin (PEC) and phycoerythrin (PE) are four major categories. Phycobiliproteins in the active conformation have the advantages of large molar extinction coefficient, large Stokes shift and strong fluorescence signal. Since the 1880s, phycobiliproteins have been used as fluorescent dyes for diagnosis, and as fluorescent dyes, they are widely used in two-color, three-color and multicolor fluorescent immunoassays. Compared with convent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/30C07K1/18
Inventor 张玉忠颜世敢张熙颖陈秀兰周百成
Owner SHANDONG UNIV
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