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Method for separating and preparing medical phycocyanin from alga

A phycocyanin, pharmaceutical-grade technology, applied in the field of protein engineering, can solve the problems of high cost and low yield of algae phycocyanin, and achieve the effects of stable properties, simple operation and cost reduction

Inactive Publication Date: 2012-10-17
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to overcome the shortcomings of low yield and high cost of extracting algae phycocyanin in the prior art, the present invention provides a method for separating and preparing pharmaceutical grade phycocyanin from algae

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Select phosphate buffer systems such as disodium hydrogen phosphate-sodium citrate, disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-citric acid, disodium hydrogen phosphate-potassium dihydrogen phosphate, etc., to make pH7.0 and concentration 0.15 mol / L buffer. Weigh 10.0 g of fresh spirulina, add 100 mL of buffer solution, and use 200 W power to ultrasonically break for 5 min with a working time of 4 s and an interval of 2 s. After the treated feed liquid was centrifuged at a high speed for 10 min at 5°C, the supernatant was absorbed, and the extract was adjusted to pH 3.5-4.5 with pharmaceutical grade HCl solution. Serum. Add 1 / 4 of the supernatant volume buffer solution to the centrifuged precipitate to fully dissolve, collect the supernatant after centrifugation, combine with the original supernatant, spray dry the obtained solution, and store the dry powder in cold storage.

[0022] CaCl with a molar concentration of 0.5 mol / L...

Embodiment 2

[0024] The acetate buffer system was selected and prepared as a buffer solution with a pH of 6.0 and a concentration of 0.10 mol / L. Weigh 10.0 g of fresh spirulina, add 10 mL of buffer solution, and use 50 W power to ultrasonically break for 30 min, with a working time of 4 s and an interval of 2 s. After the treated feed liquid was centrifuged at high speed at 4°C for 10 min, the supernatant was absorbed, and the extract was adjusted to pH 5.5-6.0 with pharmaceutical grade HCl solution. After standing for 1 h, the membrane pore size was 0.45 μm. The filter membrane is filtered, and the obtained solution is spray-dried, and the dried powder is refrigerated and stored.

[0025] CaCl with a molar concentration of 0.5 mol / L 2 solution and Na 2 HPO 4 Mix the solution into the beaker with a constant flow pump at the same flow rate of 10 mL / min, and stir at a low speed to ensure that the precipitate can be suspended in the solution, but not too fast. After standing still, pour ou...

Embodiment 3

[0027] The lactate buffer system was selected and prepared as a buffer solution with a pH of 8.0 and a concentration of 0.05 mol / L. Weigh 10.0 g of fresh spirulina, add 100 mL of buffer solution, and use 100 W power to ultrasonically break for 20 min, with a working time of 4 s and an interval of 2 s. After the treated feed liquid was centrifuged at 6°C for 10 min at low temperature, the supernatant was drawn, and the extract was adjusted to pH 4.5-5.5 with pharmaceutical grade HCl solution. After standing for 1 h, the membrane pore size was 0.20 μm The filter membrane is filtered, and the obtained solution is spray-dried, and the dried powder is refrigerated and stored.

[0028] Prepare a hydroxyapatite chromatography column, equilibrate overnight with buffer solution at a linear flow rate of 23 cm / h after filling. And use the buffer as the mobile phase, measure the elution peaks at different times in A 620 Under the absorbance value, the eluate was collected using a fracti...

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PUM

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Abstract

The invention belongs to the field of protein engineering, in particular to a method for separating and preparing medical phycocyanin from alga. The method comprises the steps of processing alga plants by an ultrasonic assistant extraction technology at low temperature, extracting alga protein by a protein extraction buffer solution to obtain a coarse extract; then adjusting pH of the coarse extract to be acidic, standing, centrifuging clear liquid and taking supernatant and adjusting pH to be neutral; or adjusting pH of the coarse extract to be acidic, performing membrane filtration after standing to obtain a filtrate; and subjecting the filtrate to hydroxyapatite column chromatography, wherein the moving phase is the protein extraction buffer solution, collecting eluting peak of phycocyanin, and then carrying out spray-drying to obtain the medical phycocyanin. The method for separating and preparing the medical phycocyanin from the alga provided by the invention overcomes the deficiency that the yield is low and the cost is high in the prior art for extracting the phycocyanin. The alga chromoprotein composite with the phycocyanin purity of A620 / A280 of more than 2.0 and the recover rate of the phycocyanin of more than 25% is obtained for the first time, and therefore, the method for separating and preparing the medical phycocyanin from the alga provided by the invention simplifies the separating steps, and is simple to operate and reduces the cost. The property of the phycocyanin is stable.

Description

technical field [0001] The invention belongs to the field of protein engineering, in particular to a method for separating and preparing pharmaceutical grade phycocyanin from algae. technical background [0002] As a protein storage unit, phycocyanin in algae has obvious anti-oxidation, anti-inflammation, enhancing the body's immunity, anti-radiation and other activities. After separation and purification, phycocyanin becomes a highly fluorescent protein, which can be used as biomedical preparations such as fluorescent probes. The existing separation and purification process of phycocyanin mainly includes precipitation, centrifugation, dialysis, chromatography and other processes. This series of methods is difficult and expensive to carry out, and the purity obtained is not high, so it is only suitable for the preparation of phycocyanin in the laboratory . [0003] Ganapathi Patil reported a simple and effective new method for the purification of C-phycocyanin, which inclu...

Claims

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Application Information

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IPC IPC(8): C07K14/795C07K1/36C07K1/34C07K1/16
Inventor 傅红闫岩
Owner FUZHOU UNIV
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