Preparation for high-purity laver phycoerythrin with one-step chromatography

A technology of laver phycoerythrin and phycoerythrin, which is applied to the preparation method of peptides, algae/moss peptides, chemical instruments and methods, etc., can solve the problems of increasing costs and achieve the effect of improving the value of deep processing

Inactive Publication Date: 2008-09-24
NANJING AGRICULTURAL UNIVERSITY
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  • Preparation for high-purity laver phycoerythrin with one-step chromatography

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Embodiment 1

[0021] (1) Weigh 4 grams of seaweed powder, soak it with PBS (1 mM, pH 6.8) buffer solution at a volume ratio of 1:50 for 1 hour, and fully swell. Cells were disrupted in an ice bath with an ultrasonic wave of 800W power for 20min. Centrifuge at 5000rmp for 20min to obtain crude phycoerythrin extract. Precipitate with ammonium sulfate with a saturation of 45%, centrifuge (5000rmp, 20min), dissolve the precipitate in 40mL, 1mmol L -1 Phosphate buffer (pH6.8);

[0022] (2) The phycoerythrin solution obtained by precipitation with ammonium sulfate with a saturation degree of 45% is first filtered through a 0.1um microfiltration membrane, and then concentrated by ultrafiltration with a membrane pore size of 10KDa. Dialyze, reverse phase precipitation with ammonium sulfate with a saturation of 20%, and centrifuge (5000rmp, 20min). Then, using an efficient crystallization method, the supernatant was precipitated with ammonium sulfate with a saturation of 65%, and the crystallizat...

Embodiment 2

[0025] (1) Weigh 4 grams of seaweed powder and soak it in PBS (1 mM, pH 6.8) buffer solution at a ratio of 1:50 for 1 hour to fully swell. Cells were disrupted in an ice bath with an ultrasonic wave of 800W power for 20min. Centrifuge at 5000rmp for 20min to obtain crude phycoerythrin extract. Precipitate with ammonium sulfate with a saturation of 45%, centrifuge (5000rmp, 20min), and dissolve the precipitate in 40mL, 1mmol·L-1 phosphate buffer (pH6.8);

[0026] (2) The phycoerythrin solution obtained by ammonium sulfate precipitation with a saturation of 45% is first filtered with a 0.1um microfiltration membrane, then concentrated by an ultrafiltration membrane with a membrane aperture of 30KDa, and then a membrane with a membrane aperture of 10KDa ultrafiltration. Dialyze, reverse phase precipitation with ammonium sulfate with a saturation of 20%, and centrifuge (5000rmp, 20min). Then, using an efficient crystallization method, the supernatant was precipitated with ammon...

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Abstract

The present invention relates to a method for preparing high-purity laver phycoerythrin by one-step chromatography, belonging to the preparation technology of the functional ingredients of halobios. Laver is swelled in PBS (1mM, pH6.8) buffer solution with a volume fifty times larger than the volume of the laver and smashed, then 800 watts of ultrasonic waves carry out cell disruption for 800 seconds to produce crude extract, which is precipitated by ammonium sulphate with 45 percent of saturation, microfiltrated via 0.1Mu m of film, ultrafiltered via a 10kDa film, goes through reversed-phase precipitation by ammonium sulphate with 20 percent of saturation and is precipitated and crystallized by ammonium sulphate with 65 percent of saturation under the temperature of 4 DEG C, and one week later, the crude extract passes through a DEAE cellulose-52 anion-exchange chromatography column, is gradiently eluted by PBS (1mM, pH6.8) buffer solutions with different concentrations of NaC1, separated and purified to produce the laver phycoerythrin. The purity (A562nm/A280nm) reaches 5.24, which accords with the reagent-grade requirement.

Description

1. Technical field [0001] The invention prepares high-purity laver phycoerythrin by a one-step chromatography method, belongs to the preparation technology of marine biological functional components, and is used for researching and developing base materials of medical health care products and functional foods. 2. Background technology [0002] Phycobiliprotein is a light-harvesting pigment protein in algal phycobilisomes, which can efficiently transfer the captured light energy to chlorophyll, so that the photosynthesis of algae can occur. Phycobiliproteins mainly include three types of phycocyanin, phycoerythrim and allophycocyanin, among which phycocyanin (PC) and phycoerythrin (PE) are more abundant in algae. Rich. The content of phycobiliprotein in algae can reach more than 28% of dry cell weight. [0003] Phycobiliprotein is a very important raw material for photodynamic drugs. Its unique photosensitizing effect can assist laser cancer treatment, enrich in the lesion,...

Claims

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Application Information

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IPC IPC(8): C07K1/18C07K14/405
Inventor 胡秋辉杨方美顾丽辛志宏刘承初周玉林赵立艳黎军胜顾卫明高军徐娟顾俊余芳徐春
Owner NANJING AGRICULTURAL UNIVERSITY
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