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Enrichment and separation method of blue green algae phycocyanin

A technology of phycocyanin and separation method, which is applied in the field of extraction and separation of high-purity phycocyanin, can solve the problems of low extraction rate, increased production cost, cumbersome process and the like, and achieves simplified steps, low production cost and strong adsorption. Effect

Active Publication Date: 2009-10-21
DONGTAI CITY SPIRULINA BIO ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reported patent technologies for preparing phycocyanin, such as Chinese patents CN1106414A, CN1130028A, CN101003565A, CN00117512.2, CN100434528C, etc., have a common feature that the extraction of phycocyanin needs to go through the stages of wall breaking, salting out or isoelectric point rough extraction , the extraction rate is not high, the production cost increases, and the process is cumbersome

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Preparation of cyanobacteria wall breaking liquid

[0020] Take the dry powder of pure spirulina produced in Dongtai, Jiangsu, use deionized water as the extractant, the solid-liquid ratio of algae powder and deionized water is 1:10, freeze at -20°C for 4 hours, and then melt at 37°C. So repeated freezing and thawing 4 times. Centrifuge the thawed cyanocyanin solution at 4800-18000r / min for 10-30min, take the supernatant, and obtain phycocyanin with a purity of A 620 / A 280 >0.41 cyanobacteria wall breaking solution.

[0021] (2) Preparation of JDN-3 type resin

[0022] In a 1L stirred tank, put 40g of chlorine balls (cross-linking degree 10%, purchased from Shanghai Jinkai Resin Co., Ltd., average particle size 340μm) and 420mL of N,N-dimethylformamide, and pass through nitrogen protection , disperse and swell for 4 hours at a stirring speed of 100r / min, add 200g of polyethylene polyamine and 0.2g of tin dichloride, heat to 70°C, and stir for 4 hours. The amin...

Embodiment 2

[0030] (1) Preparation of cyanobacteria wall breaking liquid

[0031] Take fresh microalgae produced in Taihu Lake, Wuxi, Jiangsu, add tap water as extractant, the solid-to-liquid ratio is 1:2, prepare the cyanobacteria wall breaking solution with the same steps as in Example 1, and the purity of phycocyanin is A 620 / A 280 >0.25.

[0032] (2) Preparation of JDN-3 type resin

[0033] In a 1L stirred tank, put 40g of chlorine balls (crosslinking degree 10%) and 380mL of 1,4-dioxane, pass through nitrogen protection, disperse and swell for 6 hours at a stirring speed of 100r / min, add 158 g of polyethylene polyamine and 0.08 g of magnesium oxide were heated to 80° C. and stirred for 10 hours. The aminated resin was washed sequentially with methanol, tetrahydrofuran and deionized water to neutrality, and no chloride ion was detected by 1% silver nitrate solution.

[0034] (3) Direct enrichment of phycocyanin

[0035] Add 50mL of cyanobacteria wall-breaking solution and 5g of JD...

Embodiment 3

[0041] (1) Preparation of cyanobacteria wall breaking liquid

[0042] Take fresh spirulina produced in Yancheng, Jiangsu, add tap water as an extractant, the solid-to-liquid ratio is 1:5, prepare the cyanobacteria wall breaking liquid with the same steps as in Example 1, and the purity of phycocyanin is A 620 / A 280 >0.5.

[0043] (2) Preparation of JDN-3 type resin

[0044] In a 1L stirred tank, put 40g of chlorine balls (cross-linking degree 10%) and 360mL of pyridine, pass through nitrogen protection, disperse and swell at a stirring speed of 100r / min for 8 hours, add 100g of polyethylene polyamine and 0.05 g cobalt chloride, heated to 90 ° C, stirred for 4 hours. The aminated resin was washed sequentially with methanol, tetrahydrofuran and deionized water to neutrality, and no chloride ion was detected by 1% silver nitrate solution.

[0045] (3) Direct enrichment of phycocyanin

[0046] Add 50mL of cyanobacteria wall-breaking solution and 10g of JDN-3 resin into 500mL...

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Abstract

An enrichment and separation method of blue green algae phycocyanin belongs to the technical field of biochemical separation engineering. The invention comprises the following steps: blue green algae disrupted solution is prepared; JDN-3 type resin enriches phycocyanin in the blue green algae disrupted solution; phycocyanin is eluted and separated; chromatographic separation is carried out on the JDN-3 type resin; freeze drying is carried out on eluent after desalination to obtain phycocyanin with certain purity; the recovery rate of albumen is up to 85.5%. The method is simple, directly uses the JDN-3 type resin to enrich the blue green algae disrupted solution without using usual ammonium sulfate salt precipitation, organic solvents, isoelectric points and other similar methods, reduces the dosage of chemical agents in the preparation process, simplifies the steps of the purification process, avoids the loss of products; the phycocyanin obtained by the chromatography of the JDN-3 type resin has higher purity which is up to A620 / A280>2.0. The raw material uses fresh algae or dried algae powder, or even field blue-green algae in Taihu Lake, thereby providing a technical method for solving scale use of algal resource.

Description

technical field [0001] The invention provides a simple and rapid method for extracting and separating high-purity phycocyanin from cyanobacteria. It belongs to the technical field of biochemical separation engineering. Background technique [0002] Phycocyanins are a class of pigment complex proteins present in the phycobilisomes of microalgae such as cyanobacteria and red algae. Phycobilisomes are supramolecular light-harvesting pigment complexes unique to red and cyanobacteria. Phycobilisomes are binding proteins composed of apoprotein and phycocyanin (an open-chain tetrapyrrole chromophore) covalently linked by one or two thioether bonds. The α subunit of phycocyanin contains 1 phycocyanin, and the β subunit contains 2 phycocyanins. Phycocyanin has high development and utilization value. Phycocyanin is bright and bright in color, and it is the first choice of pure natural pigment for food, high-grade eye shadow and lipstick. It can be used as a pure natural pigment in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C07K14/405C07K1/14C08F12/18C08F8/32
Inventor 王峰高瑾崔正刚
Owner DONGTAI CITY SPIRULINA BIO ENG CO LTD
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