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Extracting and purifying method of phycocyanin in hair-like seaweed

A technology of phycocyanin and purification method, which is applied in the field of natural active substance preparation, can solve the problems of phycobiliprotein denaturation, long time, and easy clogging of chromatographic columns, and achieve the effects of high processing efficiency, increased concentration and purity

Inactive Publication Date: 2016-11-23
上海宥佳医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage is that the extraction of chemical reagents can easily denature phycobiliproteins, and the hydroxyapatite column packing particles are too fine, which can easily clog the chromatography column, so only short columns can be used for purification, so it is only suitable for small-scale laboratories, and it is not easy to operate on a large scale.
Because phycoerythrin and phycocyanin have a wide range of application values, but at the same time, their extraction and purification costs are high, the time is long, and the yield is low, so that the price of this type of protein has been high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Take 50g of Fa Cai dry powder, mix the Fa Cai powder and buffer solution at a mass / volume ratio of 1:10, add 500mL of 0.05 phosphate buffer solution with pH=6.5, add 10U / g of Fa Cai powder pectinase, The comprehensive treatment makes the phycocyanin exudate to obtain the wall-breaking fluid.

[0023] 2. Centrifuge the broken wall solution to get the crude protein extract, put the crude extract on the activated carbon column and process it in series with three columns, and collect the supernatant under the column. The activated carbon used is 40-60 mesh columnar shell activated carbon; the upper column speed is 0.5BV / h, and the beading treatment starts when the light transmittance of the supernatant liquid under the column is basically the same as that of the upper column solution, and the solution with absorbance value at 280nm under the column is collected. The saturated carbon column is regenerated with 0.5% sodium hydroxide solution, and the column rate of the reg...

Embodiment 2

[0029] 1. Take 50g of Fa Cai dry powder, mix the Fa Cai powder and buffer solution at a mass / volume ratio of 1:5, add 250mL of 0.02 phosphate buffer solution with pH=6.5, add 50U / g of Fa Cai powder pectinase, The comprehensive treatment makes the phycocyanin exudate to obtain the wall-breaking fluid.

[0030] 2. Centrifuge the broken wall solution to get the crude protein extract, put the crude extract on the activated carbon column and process it in series with three columns, and collect the supernatant under the column. The activated carbon used is 20-40 mesh columnar fruit shell activated carbon; the upper column speed is 1.0BV / h, and the beading process is started when the light transmittance of the supernatant liquid under the column is basically the same as that of the upper column solution, and the solution with absorbance value at 280nm under the column is collected. The saturated carbon column is regenerated with 1% sodium hydroxide solution, and the column rate of th...

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Abstract

The invention provides an extracting and purifying method of phycocyanin in hair-like seaweed. The hair-like seaweed serving as the raw material is comprehensively treated by a phosphate buffer solution of 0.02-0.2 M and pectinase of 10-100 U / g so that phycocyanin can effuse out of cladophora cells; centrifuging is conducted to obtain a crude extraction solution, and clear liquid passes through an activated carbon column with activated carbon mesh number of 20-100 mesh and is collected; the clear liquid is subjected to salting-in and salting-out in a graded mode through ammonium sulfate of 20-70%, supernate is collected when the saturation degree is 20-35%, precipitates are collected when the saturation degree is 55-70%, and electrodialysis desalination is conducted; crude protein fluid obtained after desalination is treated by a weakly basic anion resin upper column and eluted by a phosphate buffer solution of 0.02-0.2 M in a gradient mode to collect a phycocyanin solution; a high-purity phycocyanin solution is obtained after concentration is conducted through an ultrafiltration membrane.

Description

technical field [0001] The invention relates to a method for extracting and purifying phycocyanin in Nostocs chinensis, belonging to the technical field of natural active substance preparation. Background technique [0002] Phycobiliproteins are important auxiliary pigments for algae to absorb and transmit light energy and carry out photosynthesis. There are four types of cyanocyanin (PEC), among which PE and PC are the most studied protein types. Spectral properties are one of the important characteristics of phycobiliproteins. After spectral scanning, the purity and content of phycobiliproteins can be calculated according to the measured OD value, which provides an important basis for the identification of phycobiliproteins. PE has a maximum characteristic absorption peak at 564-568nm, and a maximum fluorescence emission peak between 575nm-580nm. PC has a maximum characteristic absorption peak at 615-620nm, and a maximum fluorescence emission peak between 635-647nm. ...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/36C07K1/34C07K1/30C07K1/18
Inventor 梁靖瑞
Owner 上海宥佳医药科技有限公司
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