Extracting and purifying method for nostoc sphaeroids kutzing phycobiliprotein and purified phycoerythrin

A technology of phycoerythrin and phycobiliprotein of Radix Radix et Rhizoma, which is applied in the fields of peptide preparation, chemical instruments and methods, organic chemistry, etc., can solve the problems of market demand, complicated operation and high production cost that cannot be prepared in large quantities. , to achieve the effect of high product added value, simple purification method and low cost

Active Publication Date: 2017-09-26
HUNAN YANDI BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To separate phycoerythrin, which is the standard for commercial applications, use the reported separation and purification method of phycobiliprotein, the operation is complicated, resulting in too high production cost, and it cannot be prepared in large quantities to meet the potential market demand

Method used

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  • Extracting and purifying method for nostoc sphaeroids kutzing phycobiliprotein and purified phycoerythrin
  • Extracting and purifying method for nostoc sphaeroids kutzing phycobiliprotein and purified phycoerythrin
  • Extracting and purifying method for nostoc sphaeroids kutzing phycobiliprotein and purified phycoerythrin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Purification of phycoerythrin by using DEAE-Cellulose 52 anion exchange column in laboratory experiment

[0057] Add 50.01g, 50.03g, and 49.99g of kudzu powder into 2500mL of pure water respectively, stir well, then add 20mL of liquid nitrogen, and stir vigorously while adding. After the liquid nitrogen is completely volatilized, continue to stir until the crushed ice is completely dissolved, centrifuge, filter, and freeze-dry to obtain 10.02g, 9.98g, and 10.01g of Pueraria phycobiliprotein crude extract powders, respectively. Dissolve them in PB buffer Buffer A with a pH of 7.0 and a conductivity of 5.7 μs / cm, centrifuge at 8000 r / min for 30 minutes, and filter with a 0.2 μm filter membrane; pass through a DEAE-Cellulose 52 anion exchange column: 2) Equilibrate with 5 column volumes of PB buffer Buffer A; 3) Load 2 column volumes. Collect 40-100% of the PB-NaCl eluate with a pH of 7.0 and a conductivity of 5.7 μs / cm. The results are shown in Table 1 below: ...

Embodiment 2

[0061] Embodiment 2 adopts DEAE-Cpato anion exchange column to carry out laboratory experiment purification phycoerythrin

[0062] Add 50.11g, 50.07g, and 49.91g of Ge Xianmi dry powder to 2500mL of pure water respectively, stir well, then add 20mL of liquid nitrogen, and stir vigorously while adding. After the liquid nitrogen is completely volatilized, continue to stir until the crushed ice is completely dissolved, centrifugally filter, and freeze-dry to obtain 9.97g, 10.03g, and 10.01g of kudzu phycobiliprotein crude extract powders, respectively, with a pH of 7.0 and a conductivity of Dissolve PB buffer Buffer A at 5.7μs / cm, centrifuge at 8000r / min for 30min, and filter with a 0.2μm membrane; use DEAE-Cpato anion exchange column to pass through the column: 1) Wash the pump with 1 column volume of deionized water; 2 ) Equilibrate with 5 column volumes of PB buffer Buffer A; 3) Load 2 column volumes. Collect 40-100% of the PB-NaCl eluate with a pH of 7.0 and a conductivity o...

Embodiment 3

[0066] Example 3 Using DEAE-Sephadex Fast Flow Anion Exchange Column to Purify Phycoerythrin in Laboratory Experiment

[0067] Add 49.98g, 50.13g, and 49.92g of kudzu powder into 2500mL of pure water respectively, stir well, then add 20mL of liquid nitrogen, and stir vigorously while adding. After the liquid nitrogen is completely volatilized, continue to stir until the crushed ice is completely dissolved, centrifuge, filter, and freeze-dry to obtain 9.96g, 9.97g, and 10.02g of kudzu phycobiliprotein crude extraction powder, respectively, with a pH of 7.0 and a conductivity of Dissolve PB buffer Buffer A at 5.7μs / cm, centrifuge at 8000r / min for 30min, and filter with a 0.2μm filter membrane; use DEAE-Sephadex Fast Flow anion exchange column to pass through the column: 1) Wash the pump with 1 column volume of deionized water ; 2) equilibrate with 5 column volumes of PB buffer solution Buffer A; 3) load 1 column volume, collect 40% to 100% of the PB-NaCl eluate with a pH of 7.0 ...

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Abstract

The invention discloses a purifying method for nostoc sphaeroids kutzing phycobiliprotein. The purifying method comprises the steps: (1) choosing an anion exchange column to perform column chromatography; (2) preparing a PB buffer solution Buffer A and a PB-NaCl buffer solution Buffer B; (3) dissolving nostoc sphaeroids kutzing phycobiliprotein coarse extract powder through the PB buffer solution Buffer A, centrifuging and filtering; (4) washing a pump and utilizing the PB buffer solution Buffer A to balance an ion exchange column; (5) directly loading sample of the nostoc sphaeroids kutzing phycobiliprotein which is prepared in the step (3) and dissolved into the PB buffer solution Buffer A, performing gradient elution through the PB-NaCl buffer solution Buffer B and collecting eluant; (6) utilizing a saline solution to clean the iron exchange column and reservedly cleaningback flushing through an alkali solution; (7) performing ultrafiltration and concentration on leacheate and dialyzing to obtain a purified nostoc sphaeroids kutzing phycobiliprotein water solution. The method disclosed by the invention is simple to operate, the concentration and the purity of the obtained nostoc sphaeroids kutzing phycobiliprotein are both obviously improved, and the method is suitable for industrial large-scale production.

Description

technical field [0001] The invention relates to a method for efficiently extracting and purifying the phycoerythrin of Pueraria japonica and the purified phycoerythrin. Background technique [0002] Nostoc sphaeroids Kutzing (Nostoc sphaeroids Kutzing), scientific name Nostoc sphaeroides, belongs to Nostoc. Fungus (Nostoc commune vauch) is a precious nitrogen-fixing cyanobacteria traditionally exported in my country. It is a multicellular filamentous plant with a simple cell structure. The individual is composed of spherical cells and unbranched single-row filaments. The filaments are bead-shaped, and the groups are colloidal, spherical or other irregular shapes. , blue-green or yellowish-brown, visible to the naked eye, has the ability to fix nitrogen. "Compendium of Materia Medica", "Medicinal Tests", "National Chinese Herbal Medicine Compilation" and so on claim that the effect of Gexianmi is to improve eyesight and nourish qi, make people have children, relieve heat and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/405C07K1/36C07K1/18C07K1/34C07K1/14
CPCC07K14/405
Inventor 余佳张瑞华王玉兰占豪陈盛王维
Owner HUNAN YANDI BIOLOGICAL ENG
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