Method for separating and purifying phycoerythrin from bangia fusco-purpurea

A technology for separation and purification of phycoerythrin, applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve the problems of cumbersome separation and purification process, complicated operation, and high cost, so as to save sample loading time and simplify purification Steps, the effect of accelerating the desalination speed

Inactive Publication Date: 2011-11-16
曹敏杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the separation and purification of phycoerythrin is mainly carried out by ammonium sulfate fractional precipitation, ion exchange column chromatography, hydro

Method used

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  • Method for separating and purifying phycoerythrin from bangia fusco-purpurea
  • Method for separating and purifying phycoerythrin from bangia fusco-purpurea

Examples

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Embodiment 1

[0019] 1. Extraction: take 100mL of distilled water to swell 5g of red hair algae powder, after freezing and thawing, add 200mL of distilled water for stirring, mash the tissue, ultrasonically crush, then filter and centrifuge, and collect about 300mL of the supernatant, which is the crude phycoerythrin Extraction.

[0020] 2. Salting out: Add 58.20 g of solid ammonium sulfate to 300 mL of crude extract to make the solution reach 35% saturation of ammonium sulfate, let it stand for 2 hours, and centrifuge to discard the precipitate. Add 29.14g of ammonium sulfate to the supernatant to 50% saturation, let it stand for 2 hours, discard the supernatant after centrifugation, and dissolve the precipitate with 0.02mol / L phosphate buffer (pH=7.0), the volume is about 100mL. Further use the 30kDa molecular weight cut-off ultrafiltration membrane to concentrate and desalt and remove impurity proteins with a molecular weight less than 30kDa. During the concentration process, appropriat...

Embodiment 2

[0026] 1. Extraction: Take 200mL of distilled water to swell 10g of red hair algae powder, after freezing and thawing, add 550mL of distilled water for stirring, mash the tissue, ultrasonically crush, then filter and centrifuge, and collect about 750mL of the supernatant, which is the crude phycoerythrin Extraction.

[0027] 2. Salting out: Add 145.50 g of solid ammonium sulfate to 750 mL of the crude extract to make the solution reach 35% saturation of ammonium sulfate, let it stand for 2 hours, discard the precipitate after centrifugation, and add 71.78 g of ammonium sulfate to the supernatant to 50 % saturation, let it stand for 2 hours, discard the supernatant after centrifugation, and dissolve the precipitate with 0.02mol / L phosphate buffer (pH=7.0) to obtain a salting-out solution with a volume of about 180mL. Further use the 30kDa molecular weight cut-off ultrafiltration membrane to concentrate and desalt and remove impurity proteins with a molecular weight less than 30...

Embodiment 3

[0032] 1. Extraction: take 500mL of distilled water to swell 20g of red hair algae powder, after freezing and thawing, add 1000mL of distilled water to fully stir, mash the tissue, ultrasonically crush, then filter and centrifuge, and collect about 1500mL of the supernatant, which is phycoerythrin crude extract.

[0033] 2. Salting out: Add 291g of solid ammonium sulfate to 1500mL crude extract to make the solution reach 35% saturation. After standing for 2 hours, centrifuge at 8000g to discard the precipitate, and add 143.56g of ammonium sulfate to the supernatant to 50% saturation After centrifugation, the supernatant was discarded, and the precipitate was dissolved with 0.02mol / L phosphate buffer (pH=7.0) to obtain a crude phycoerythrin salting-out solution with a volume of about 260mL. Further use the 30kDa molecular weight cut-off ultrafiltration membrane to concentrate and desalt and remove impurity proteins with a molecular weight less than 30kDa. During the concentrat...

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Abstract

The invention discloses a method for separating and purifying phycoerythrin from bangia fusco-purpurea. According to the method, foreign proteins with small molecular weights are removed at the same time of salt removal by a membrane technology, and the phycoerythrin is effectively separated from the foreign proteins based on the principles that the bonding strength between anion exchange resin and the phycoerythrin is obviously changed in solutions of different salt contents and different pH values, and the like, thereby obtaining high-purity and high-yield phycoerythrin. In different components, the purity of the phycoerythrin is generally higher than 3.0 (commonly accepted standard) and is 5.5 at most, and the yield is about 0.8g/100g bangia fusco-purpurea. The method has a simple process and good separating effect, and realizes quick and effective separation and purification of the phycoerythrin.

Description

technical field [0001] The invention discloses a method for extracting phycoerythrin, in particular to a method for separating and purifying phycoerythrin in red algae. Background technique [0002] Phycoerythrin is an important light-harvesting pigment protein of many seaweeds. It forms rod-shaped phycobilisomes with phycobiliproteins such as phycocyanin and isophycocyanin. Phycobilisomes can capture and transmit light energy. Phycoerythrin is mainly distributed in the red algae of seaweeds. Due to the existence of pigment molecules, it has strong absorption in the 480-570nm band of visible light and can produce strong fluorescence. Phycoerythrin can be used as a natural pigment in food, cosmetics, dyes and other industries, and can also be made into fluorescent reagents for clinical medical diagnosis and research fields such as immunochemistry and bioengineering. Phycoerythrin is also an important physiologically active substance, which can be made into a photosensitizer ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/36C07K1/34C07K1/30C07K1/18
Inventor 曹敏杰付晓苹刘光明苏文金
Owner 曹敏杰
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