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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  • 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 phycocyan

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0025] Example 1:

[0026] 1 Preparation of hydroxyapatite:

[0027] (1) Solution preparation: separately prepare 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 (containing 0.1M sodium chloride, pH=6.7) Containing 0.1M sodium chloride, pH=6.7);

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

[0029] (3) Preparation of calcium hydrogen phosphate precipitate: Under constant stirring of a glass rod, mix 2M dipotassium hydrogen phosphate solution and 2M calcium chloride solution in equal volumes, and slowly drip the mixture into the container at a speed of 3 ml / min. To form a calcium hydrogen phosphate precipitate;

[0030] (4) Transform into hydroxyapatite: Under continuous stirring, add 2M potassium hydroxide solution dropwise to the above calcium hydrogen phosphate precipitation. When the pH value of the mi...

Example Embodiment

[0037] Example 2:

[0038] 1 Preparation of hydroxyapatite:

[0039] (1) Solution preparation: respectively prepare 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 (containing 0.15M sodium chloride, pH=6.9) Containing 0.15M sodium chloride, 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 precipitate: Under constant stirring of a glass rod, mix 1.5M dipotassium hydrogen phosphate solution and 1.5M calcium chloride solution in equal volumes, and slowly drip the mixture into the container at a speed of 5 ml / min. To form a calcium hydrogen phosphate precipitate;

[0042] (4) Converted into hydroxyapatite: Under continuous stirring, add 1.5M potassium hydroxide solution dropwise to the above calcium hydrogen phosphate precipitate, when the pH va...

Example Embodiment

[0049] Example 3:

[0050] 1 Preparation of hydroxyapatite:

[0051] (1) Solution preparation: respectively prepare 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 ( Containing 0.2M sodium chloride, pH=7.0);

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

[0053] (3) Balance: balance the purchased and soaked hydroxyapatite with 0.002M phosphate buffer (containing 0.2M sodium chloride, pH=7.0) to remove fine particles for chromatography.

[0054] 2 Prepare phycocyanin and allophycocyanin with a 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), where the ratio of spirulina algae powder to water is controlled at 1:15; after stirring even...

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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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