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Separation technology for aqueous two-phase extraction of spirulina phycocyanin from polyvinylether phenylacetamide

A polyvinyl ether and phenylacetamide technology, which is applied in the fields of depsipeptides, peptides, chemical instruments and methods, can solve the problems of high price, poor selectivity of separating phycocyanin, and complicated process, and achieves low production cost and solves the problem of utilization. and high-value problems, the effect of high extraction selectivity

Inactive Publication Date: 2017-02-01
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 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 1:50, 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.

[0019] (2) Preparation of polyvinyl ether mesylate

[0020] Put 100g of polyethylene glycol PEG-1000 (purchased from Haian Petrochemical Plant) and 300ml of methylene sulfonyl chloride (purchased from Jinan Zi'an Chemical Co., Ltd.) into a 1000ml round bottom flask (purchased from Dongying Huibao) Che...

Embodiment 2

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

[0029] 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.

[0030] (2) Preparation of polyvinyl ether mesylate

[0031] Put 100g of polyethylene glycol PEG-3000 (purchased from Haian Petrochemical Plant) and 300ml of methylene sulfonyl chloride (purchased from Jinan Zi'an Chemical Co., Ltd.) into a 1500ml round bottom flask (purchased from Dongying Huibao) Chemical Co., Ltd.) solution, the weight...

Embodiment 3

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

[0040] Take the spirulina powder produced in the spirulina base in Chenghaiyuan, Lijiang, Yunnan, use 100mmol / 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 is 0.72, and the protein concentration of the broken wall fluid is 5.6 mg / ml mg / ml.

[0041] (2) Preparation of polyvinyl ether mesylate

[0042] Put 100g of polyethylene glycol PEG-3000 (purchased from Haian Petrochemical Plant) and 300ml of methylene sulfonyl chloride (purchased from Jinan Zi'an Chemical Co., Ltd.) into a 2000ml round bottom flask (purchased from Dongyi...

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Abstract

The invention relates to a separation technology for aqueous two-phase extraction of spirulina phycocyanin from polyvinylether phenylacetamide, and belongs to the technical field of biochemical separation engineering. The separation technology comprises the steps of polyvinylether phenylacetamide preparation; two-phase extraction of phycocyanin; phycocyanin powder obtaining after concentration and freeze drying, wherein the protein recovery rate can reach 70 to 85 percent. The method is simple; spirulina wall breaking liquid is subjected to enrichment and separation by directly using the polyvinylether phenylacetamide; the purification process step is simplified; the selectivity is high; the spectrum purity of the phycocyanin obtained through separation achieves the effect that A620 / A280 is greater than 2.5. Raw materials can use fresh spirulina or dry spirulina powder, so that a technology method for realizing high-value large-scale utilization of spirulina resources is provided.

Description

technical field [0001] The invention provides a preparation method of polyvinyl ether phenylacetamide, which is used for extracting 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 in research fields ...

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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