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

A technology of polyvinyl ether carboxylate and phycocyanin, which is applied in the field of biochemical separation engineering, can solve the problems of high price, poor selectivity of separating phycocyanin, and complicated process, achieves low production cost, solves the problem of utilization and high value. problem, selective effect

Inactive Publication Date: 2015-04-01
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 algal phycocyanin, such as Chinese patent CN103880950A, uses expensive ionic liquids to form a two-water phase, which increases production costs. CN102993297A, CN101891809A, etc. use commercially available PEG Composition of polymer two-phase aqueous phase, the selectivity of separating phycocyanin is poor, it needs to be used in combination 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 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 1:50, and freeze at -20 °C for 4 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 1.0mg / ml, phycocyanin purity A 620 / A 280 It is 0.72mg / ml.

[0018] (2) Preparation of polyvinyl ether carboxylate

[0019] Put 100g of polyethylene glycol PEG-1000 (purchased from Haian Petrochemical Plant) and 5g of sodium block (purchased from Nanjing Chemical Reagent Co., Ltd.) in a 1000ml round bottom flask, and react for 5 hours at 25°C and magnetic stirri...

Embodiment 2

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

[0026] Take fresh spirulina produced in Jiangsu Dafengci Century Biotechnology Co., Ltd., use 50mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:100, 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 0.5.

[0027] (2) Preparation of polyvinyl ether carboxylate

[0028] Put 100g of polyethylene glycol PEG-3000 (purchased from Haian Petrochemical Plant) and 3.5g of sodium block (purchased from Nanjing Chemical Reagent Co., Ltd.) in a 1000ml round bottom flask, and react at 35°C under magnetic stirring conditions for 7 hour, in the reaction mi...

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 150mmol / L phosphate buffer as the extractant, the solid-liquid ratio of spirulina to deionized water is 1:150, and freeze at -20°C for 4 hours Finally, thaw at 37°C, and repeat the freezing and thawing process 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 was 0.77, and the protein concentration of the broken wall fluid was 4.6mg / ml.

[0036] (2) Preparation of polyvinyl ether carboxylate

[0037] In a 1000ml round-bottomed flask, put 100g of polyethylene glycol PEG-4000 (purchased from Haian Petrochemical Factory) and 5g of sodium block (purchased from Nanjing Chemical Reagent Co., Ltd.), and react at 45°C under magnetic st...

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Abstract

The invention provides a technology for separating phycocyanin in spirulina by using polyvinyl ether carboxylate through two aqueous phase extraction, belonging to the technical field of biochemical separation engineering. The technology comprises the following steps: preparing polyvinyl ether carboxylate; 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 70-90%. The technology has the beneficial effects that the method is simple; the steps of a purification process are simplified by enriching and separating broken wall liquids of the spirulina by directly using polyvinyl ether carboxylate; the selectivity is high; the spectral purity of phycocyanin obtained through separation is A620 / A280>2.7; the powder of fresh or dry spirulina can be adopted as the 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 carboxylate, which is used for extracting phycocyanin from spirulina broken wall serum. 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 in research fields such as...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/36C07K1/34C07K1/14
CPCC07K14/795
Inventor 张杜炎石敏覃晓雨韩震王峰
Owner JIANGNAN UNIV
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