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Technology for separating phycocyanin in spirulina by using polyvinyl ether alcohol amine through two aqueous phase extraction

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

Inactive Publication Date: 2015-03-04
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

[0016] (1) Preparation of spirulina broken wall serum

[0017] Take the dry powder of Spirulina platensis produced in Cibainian Bioengineering Co., Ltd., Dongtai City, Jiangsu Province, use deionized water as the extractant, and the solid-liquid ratio of algae powder to deionized water is 1:55, and freeze at -20 ° C for 3 After one hour, it was thawed at 37°C, and so repeated freezing and thawing 4 times, the thawed cyanobacteria broken wall liquid was centrifuged at 6000 rpm for 10 minutes, and the supernatant was taken as the broken wall supernatant, and the protein concentration in the supernatant was 1.2mg / ml, phycocyanin purity A 620 / A 280 0.75mg / ml.

[0018] (2) Preparation of polyvinyl ether alcohol amine

[0019] In the round bottom flask of 1000ml, put into 100g polyethylene glycol PEG-500 (purchased from Haian Petrochemical Plant), the average molecular weight of PEG-500 is 500, in the constant pressure funnel, pack 50g phosphorus tribromide (purchased from Huaia...

Embodiment 2

[0025] (1) Preparation of spirulina wall breaking liquid

[0026] Take fresh spirulina produced in Jiangsu Dafengci Century Biotechnology Co., Ltd., use 100mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:80, and freeze at -10°C After 4 hours, it was thawed at 37°C and repeated freezing and thawing 5 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. The protein concentration in the supernatant was 3.1 mg / ml, and the purity of phycocyanin was A 620 / A 280 is 0.55.

[0027] (2) Preparation of polyvinyl ether alcohol amine

[0028] Put 100g polyethylene glycol PEG-2000 (purchased from Hai'an Petrochemical Plant) into the round bottom flask of 1000ml, the average molecular weight of PEG-3000 is 2000, in the constant pressure funnel, pack 35g phosphorus tribromide (purchased from Huaian City Xinxin Che...

Embodiment 3

[0034] (1) Preparation of spirulina wall breaking liquid

[0035] Take the spirulina powder produced in the spirulina base in Chenghaiyuan, Lijiang, Yunnan, use 50mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:100, and freeze at -20°C for 5 hours Finally, thaw at 37°C, and repeat the freezing and thawing process 5 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 4.6 mg / ml mg / ml.

[0036] (2) Preparation of polyvinyl ether alcohol amine

[0037] 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-4000 is 4000g every mole, in the constant pressure funnel, lo...

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Abstract

The invention provides a technology for separating phycocyanin in spirulina by using polyvinyl ether alcohol amine through two aqueous phase extraction, belonging to the technical field of biochemical separation engineering. The technology comprises the following steps: preparing polyvinyl ether alcohol amine; carrying out two aqueous phase extraction on phycocyanin; and obtaining phycocyanin powder after concentrating and freeze-drying a phycocyanin aqueous solution, wherein the phycocyanin recovery rate can be 75-87%. The technology has the beneficial effects that the method is simple; the steps of a purification process are simplified by enriching and separating the spirulina broken wall liquid by directly using polyvinyl ether alcohol amine; the selectivity is high; the spectral purity of phycocyanin obtained through separation is A620 / A280 which is greater than 2.4; the powder of fresh or dry spirulina can be adopted as a raw material, thus providing a technical method with the effect of solving the problem of high value and large scale utilization of spirulina resources.

Description

technical field [0001] The invention provides a preparation method of polyvinyl ether alcohol amine, which is used for two-phase aqueous extraction of phycocyanin from spirulina wall breaking liquid. It belongs to the technical field of biochemical separation engineering. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C07K1/14C08G65/00
CPCC07K14/795C08G65/00C08G2650/02
Inventor 宋佳玉张杜炎石敏覃晓雨韩震王峰
Owner JIANGNAN UNIV
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