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Separation technique of Spirulina phycocyanin by polyvinylether aniline double-water-phase extraction

A polyvinyl ether aniline and phycocyanin technology, which is applied in the directions of depsipeptides, peptides, chemical instruments and methods, can solve the problems of high price, cumbersome process, poor selectivity for separating phycocyanin, etc., and achieves low production cost and high selectivity. High performance, the effect of solving the problem of utilization and high value

Inactive Publication Date: 2017-02-08
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reported patented technology of two-phase extraction and separation of phycocyanin for the preparation of algae phycocyanin, such as Chinese patent CN103880950A, uses expensive ionic liquids to form a polymer-water phase, which increases production costs. CN102993297A, CN101891809A, etc. use commercially available PEG constitutes a polymer-water phase, and the selectivity of separating phycocyanin is poor. It needs to be combined with other extraction methods such as salting out to obtain high-purity products, and the process is cumbersome.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Preparation of spirulina broken wall serum

[0018] Take the dry powder of Spirulina platensis produced in Cibaiian Biological Engineering Co., Ltd., Dongtai City, Jiangsu Province, use deionized water as the extraction agent, and the solid-liquid ratio of algae powder to deionized water is 1:65, and freeze at -20 ° C for 6 After one hour, it was thawed at 40°C, and so repeated freezing and thawing 4 times, the thawed cyanobacteria broken wall liquid was centrifuged at 6000 rpm for 10 minutes with a centrifuge, and the supernatant was taken as the broken wall supernatant, and the protein concentration in the supernatant was 2.0mg / ml, phycocyanin purity A 620 / A 280 It is 0.72mg / ml.

[0019] (2) Preparation of brominated polyvinyl ether

[0020] In the round bottom flask of 500ml, put into 100g polyethylene glycol PEG-1000 (purchased from Hai'an Petrochemical Plant), the average molecular weight of PEG-1000 is 1000g every mole, in the constant pressure funnel, loa...

Embodiment 2

[0028] (1) Preparation of spirulina wall breaking liquid

[0029] Take fresh spirulina produced in Dongtai City, Jiangsu Cibainian Bioengineering Co., Ltd., use 500mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:100, and freeze at -10°C After 7 hours, it was thawed at 37°C and repeated freezing and thawing 5 times. The thawed cyanobacteria broken wall liquid was centrifuged at 6000 rpm for 10 minutes with a centrifuge, and the supernatant was taken as the broken wall supernatant, the protein concentration in the supernatant was 5.1mg / ml, and the purity of phycocyanin was A 620 / A 280 is 0.6.

[0030] (2) Preparation of brominated polyvinyl ether

[0031] In the round bottom flask of 1000ml, put into 100g polyethylene glycol PEG-4000 (purchased from Hai'an Petrochemical Plant), the average molecular weight of PEG-3000 is 3000g every mole, load 40g phosphorus tribromide (purchased in constant pressure funnel) From Huai’a...

Embodiment 3

[0039] (1) Preparation of spirulina wall breaking liquid

[0040] Take fresh spirulina produced in Dongtai City, Jiangsu Cibainian Bioengineering Co., Ltd., use 100mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:120, and freeze at -20°C After 4 hours, it was thawed at 37°C and repeated freezing and thawing 6 times. The thawed cyanobacteria wall-broken liquid was centrifuged at 6000 rpm for 10 minutes with a centrifuge, and the supernatant was taken as the wall-broken supernatant, and the purity of phycocyanin in the supernatant was A 620 / A 280 is 0.72, and the protein concentration of the broken wall fluid is 7.6mg / ml.

[0041] (2) Preparation of brominated polyvinyl ether

[0042] In the round-bottomed flask of 2000ml, put into 100g polyethylene glycol PEG-4000 (purchased from Hai'an Petrochemical Plant), the average molecular weight of PEG-46000 is 4000g every mole, in constant pressure funnel, load 32g phosphorus tr...

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Abstract

The invention relates to a separation technique of Spirulina phycocyanin by polyvinylether aniline double-water-phase extraction, belonging to the technical field of drug separation. The technique comprises the following steps: preparing polyvinylether aniline; carrying out two-phase extraction on phycocyanin; and carrying out concentration and freeze-drying to obtain phycocyanin powder. The protein recovery rate can reach 70-95%. The method is simple: the Spirulina wall breaking solution is directly enriched and separated by using the polyvinylether aniline, thereby simplifying the steps of the purification process. The method has high selectivity; and the spectral purity of the separated phycocyanin reaches A620 / A280>3.5. The raw material can adopt Spirulina fresh algae or dry algae powder, thereby providing a high-value large-scale utilization technique of Spirulina resources.

Description

technical field [0001] The invention provides a preparation method of polyvinyl ether aniline, which is used for extracting phycocyanin from spirulina wall breaking liquid. The invention belongs to the technical field of drug separation. Background technique [0002] Spirulina phycocyanin is present in the phycobilisomes of Spirulina. Phycobilisomes are supramolecular light-harvesting pigment complexes unique to red and cyanobacteria. Phycocyanin is bright blue in color and bright in color. It is the first choice of pure natural pigment for food, high-end eye shadow and lipstick. Phycocyanin can also regulate and synthesize a variety of important enzymes required by human metabolism, has obvious anti-oxidation and anti-inflammatory effects, regulates the human immune system, and enhances the function of the immune system. High-purity phycocyanin has strong fluorescence, and it is made of pure natural fluorescent reagents, which are used in research fields such as clinical...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/14
CPCC07K14/795
Inventor 王峰
Owner JIANGNAN UNIV
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