Method for fast separating and purifying C-phycocyanin and isophycocyanin from blue algae

A technology for isophycocyanin and phycocyanin, which is applied in the field of rapid separation and purification of C-phycocyanin and isophycocyanin, can solve the problem of high prices for the development of universal diagnostic reagents and application of phycobiliprotein, and achieves a scale that is difficult to overcome. Chemical rapid preparation, simple equipment requirements, good application prospects and economic benefits

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

AI Technical Summary

Problems solved by technology

The main factor affecting the development and application of popular diagnostic reagents is the high price of phycobiliprotein

Method used

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  • Method for fast separating and purifying C-phycocyanin and isophycocyanin from blue algae
  • Method for fast separating and purifying C-phycocyanin and isophycocyanin from blue algae
  • Method for fast separating and purifying C-phycocyanin and isophycocyanin from blue algae

Examples

Experimental program
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Embodiment 1

[0032] The method for rapidly separating and purifying C-phycocyanin and isophycocyanin from cyanobacteria, the steps are as follows:

[0033] (1) Extraction of C-phycocyanin and isophycocyanin:

[0034] Take fresh spirulina, grind it with liquid nitrogen according to the weight-to-volume ratio of 1:1 (g / ml), then add 0.02mol / L phosphate buffer (pH5.8) according to the weight-to-volume ratio of fresh algae to dissolve , centrifuge at 8000rpm for 15min at 4°C, and the supernatant is the crude extract of C-phycocyanin and isophycocyanin.

[0035] (2) Preliminary purification of C-phycocyanin and isophycocyanin:

[0036] Add solid ammonium sulfate to the crude extract of C-phycocyanin and isophycocyanin obtained in the above step (1) to a concentration of 60% (w / v), let it stand for 5 hours, and then centrifuge at 10,000 rpm at 4°C After 10 minutes, the precipitate was taken and dissolved in 20 mM phosphate buffer (pH 5.8), and the phosphate buffer in which the precipitate was di...

Embodiment 2

[0040] The method for rapidly separating and purifying C-phycocyanin and isophycocyanin from cyanobacteria, the steps are as follows:

[0041] (1) Extraction of C-phycocyanin and isophycocyanin:

[0042] Take fresh Synechocystis, add liquid nitrogen to grind at a weight-to-volume ratio of 1:1 (g / ml), then add 0.02mol / L phosphate buffer (pH6.0) at a weight-to-volume ratio of 1:1 to dissolve, Centrifuge at 10,000 rpm for 10 min at 4°C, and the supernatant is the crude extract of C-phycocyanin and isophycocyanin.

[0043] (2) Preliminary purification of C-phycocyanin and isophycocyanin:

[0044] Add solid ammonium sulfate to the C-phycocyanin and isophycocyanin crude extracts prepared in the above step (1) to a concentration of 55% (w / v), leave it for 6h, and then centrifuge at 4°C at 8000rpm After 15 minutes, the precipitate was taken and dissolved in 20 mM phosphate buffer (pH 6.0), and the phosphate buffer in which the precipitate was dissolved was dialyzed, and the dialysat...

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Abstract

A rapid separation and purification process of C-phycocyanin and allophycocyanin from blue algae, which pertains to separation and purification of phycocyanin technical field. The invention extracts C-phycocyanin and allophycocyanin by freeze dissolving and intensified swelling with low ions, finally carries out primary purification after precipitation with ammonia sulfate, elutes buffer liquid with constant ionic strength and pH gradient by DEAE Sepharose Fast Flow ionexchange chromatography, so that C-phycocyanin and allophycocyanin with high purity is obtained by one step. The process is easy in operation, time and energy saving, with little requirements to apparatus, high in yield. The process also dramatically lower preparation cost of CPC and APC, thus lays a foundation for CPC and APC application in ultrasensitive detection in biomedical.

Description

technical field [0001] The invention relates to a method for quickly separating and purifying C-phycocyanin and isophycocyanin from cyanobacteria, and belongs to the technical field of phycocyanin separation and purification. Background technique [0002] In order to study the cellular localization, interaction and dynamic changes of proteins and other biomacromolecules, researchers urgently need new technologies and materials to realize the "identification", "reading" and "query" of proteins and other biomacromolecules. Currently commonly used fluorescent labels are far from being suitable for high-throughput specific labeling of biological macromolecules due to the limitations of the fluorescent properties of fluorescent dye molecules (broad fluorescence spectrum and low quantum yield). [0003] Phycobiliproteins are a kind of photosynthetic light-harvesting pigment-protein complexes, mainly found in cyanobacteria, red algae, cryptophyta and a few dinoflagellates. It play...

Claims

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Application Information

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