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

Abstract

The invention provides a technology for separating phycocyanin in spirulina by using polyvinyl ether phosphate through two aqueous phase extraction, belonging to the technical field of biochemical separation engineering. The technology comprises the following steps: preparing polyvinyl ether phosphate; carrying out two phase extraction on phycocyanin; concentrating and freeze-drying, thereby obtaining phycocyanin powder, wherein the phycocyanin recovery rate can be 70-85%. 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 phosphate; the selectivity is high; the spectral purity of phycocyanin obtained through separation is A620 / A280 which is greater than 2.5; 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 phosphate, 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 such a...

Examples

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 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, load 30g phosphorus tr...

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 brominated polyvinyl ether

[0031] In the round bottom flask of 1000ml, put into 100g polyethylene glycol PEG-3000 (purchased from Hai'an Petrochemical Plant), the average molecular weight of PEG-3000 is 3000g every mole, load 50g phosphorus tribromide (purchased in constant pressure funnel) From Huai’an Xinxin...

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 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-4000 is 4000g every mole, in constant pressure funnel, load 30g...

technical field

[0001] The invention provides a preparation method of polyvinyl ether phosphate, 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 such a...

Examples

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 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, load 30g phosphorus tr...

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 brominated polyvinyl ether

[0031] In the round bottom flask of 1000ml, put into 100g polyethylene glycol PEG-3000 (purchased from Hai'an Petrochemical Plant), the average molecular weight of PEG-3000 is 3000g every mole, load 50g phosphorus tribromide (purchased in constant pressure funnel) From Huai’an Xinxin...

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 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-4000 is 4000g every mole, in constant pressure funnel, load 30g...