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Method for preparing phycocyanin and allophycocyanin at same time

A technology for allophycocyanin and phycocyanin, which is applied in the field of simultaneous preparation of phycocyanin and allophycocyanin, can solve the problems of complex separation methods, achieve the effects of high purity, reduce production costs, and simplify cumbersome steps

Inactive Publication Date: 2007-07-25
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

"A Simple Method for Large-scale Extraction of Phycocyanin" (200510059188.8) by the Institute of Botany, Chinese Academy of Sciences is a method for large-scale extraction of phycocyanin from cyanobacteria and blue-green algae with microbial culture fluid. Advanced, but involves the cultivation of several strains, and the separation method is relatively complicated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1 Preparation of hydroxyapatite:

[0027] (1) Solution preparation: prepare respectively 0.001M phosphate buffer (containing 0.1M sodium chloride, pH=6.7), 0.025M phosphate buffer (containing 0.1M sodium chloride, pH=6.7) and 0.045M phosphate buffer ( Contains 0.1M NaCl, pH=6.7);

[0028] (2) prepare 2M dipotassium hydrogen phosphate solution and 2M calcium chloride solution respectively;

[0029] (3) Preparation of calcium hydrogen phosphate precipitation: under continuous stirring with a glass rod, mix equal volumes of 2M dipotassium hydrogen phosphate solution and 2M calcium chloride solution, and slowly drop the mixed solution into the container at a speed of 3 ml / min. Make it form calcium hydrogen phosphate precipitate;

[0030] (4) Converting into hydroxyapatite: under continuous stirring, add dropwise 2M potassium hydroxide solution to the above-mentioned calcium hydrogen phosphate precipitation, when the pH value of the mixed solution reaches 11.0, slowly stir...

Embodiment 2

[0038] 1 Preparation of hydroxyapatite:

[0039] (1) Solution preparation: prepare respectively 0.0015M phosphate buffer (containing 0.15M sodium chloride, pH=6.9), 0.03M phosphate buffer (containing 0.15M sodium chloride, pH=6.9) and 0.05M phosphate buffer ( Contains 0.15M NaCl, pH=6.9);

[0040] (2) Prepare 1.5M dipotassium hydrogen phosphate solution and 1.5M calcium chloride solution respectively;

[0041] (3) Preparation of calcium hydrogen phosphate precipitation: under continuous stirring with a glass rod, mix equal volumes of 1.5M dipotassium hydrogen phosphate solution and 1.5M calcium chloride solution, and slowly drop the mixed solution into the container at a speed of 5 ml / min In, make it form calcium hydrogen phosphate precipitation;

[0042] (4) Transformation into hydroxyapatite: under continuous stirring, dropwise add 1.5M potassium hydroxide solution to the above calcium hydrogen phosphate precipitation, when the pH value of the mixed solution reaches 11.0, ...

Embodiment 3

[0050] 1 Preparation of hydroxyapatite:

[0051] (1) Solution preparation: prepare respectively 0.002M phosphate buffer (containing 0.2M sodium chloride, pH=7.0), 0.035M phosphate buffer (containing 0.2M sodium chloride, pH=7.0) and 0.06M phosphate buffer ( Contains 0.2M NaCl, pH=7.0);

[0052] (2) Buy hydroxyapatite from the market and soak it in 0.002M phosphate buffer solution (containing 0.2M sodium chloride, pH=7.0) for one day;

[0053] (3) Equilibration: The purchased and soaked hydroxyapatite was equilibrated with 0.002M phosphate buffer solution (containing 0.2M sodium chloride, pH=7.0) to remove fine particles for chromatography.

[0054] 2 Preparation of phycocyanin and allophycocyanin with hydroxyapatite chromatography column:

[0055] (1) Preparation of phycobiliprotein crude extract: take spirulina algae powder and dissolve it in distilled water (containing 0.2M sodium chloride), wherein the ratio of spirulina algae powder to water is controlled at 1:15; Freez...

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PUM

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Abstract

This invention provides a method for preparing phycocyanin and allophycocyanin simultaneously. The method comprises: (1) preparing crude extract of phycocyanin; (2) filling a column with hydroxyapatite, and balancing the chromatographic column; (3) eluting phycocyanin and allophycocyanin, respectively; (4) concentrating, desalting and freeze-drying the eluate to obtain phycocyanin and allophycocyanin. The method utilizes powder of artificially bred Spirulina platensis as the raw material, which can be purchased in a large quantity, and has low cost and high purity. Besides, the method adopts hydroxyapatite column chromatography to separate phycocyanin and allophycocyanin, thus simplifying the preparation process. Hydroxyapatite can be easily prepared, thus lowering the cost. The method is ideal for preparing phycocyanin and allophycocyanin simultaneously.

Description

Technical field [0001] The invention provides a method for simultaneously preparing phycocyanin and allophycocyanin, belonging to the technical field of bioengineering. Background technique [0002] Phycobiliproteins are light-harvesting pigment proteins in red algae, blue-green algae, and cryptoalgae. According to their absorption spectrum properties, they can be divided into phycoerythrin (Phycoerythrin, PE), phycoerythrocyanin (Phycoerythrocyanin, PEC), and phycocyanin. Protein (Phycocyanin, PC) and allophycocyanin (Allophyxoxyanin, APC) four. Phycobiliprotein efficiently transfers the captured light energy to chlorophyll, so that photosynthesis of algae can take place. Each molecule of phycobiliprotein contains α and β subunits, the molecular weight of the subunits is about 17~22kD, (B- and R-phycoerythrin also contain γ subunit, its molecular weight is about 30kD) each polypeptide Chains contain one or more covalently linked chromophores. Scientific research in recen...

Claims

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Application Information

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IPC IPC(8): C07K1/16C07K2/00C07K14/405
Inventor 孟春晓高政权
Owner SHANDONG UNIV OF TECH
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