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Spirulina phycocyanin and extraction method thereof

A technology of phycocyanin and extraction method, which is applied to the preparation methods of peptides, chemical instruments and methods, depsipeptides, etc., can solve the problems of low product yield, long elution time, long operation period, etc., and achieve good anti-oxidation Free radical scavenging effect and fluorescence intensity, simple extraction process, low cost effect

Active Publication Date: 2013-03-27
丽江美之源食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high amino acid sequence homology between the phycocyanin molecule and the subunit of the allophycocyanin molecule in spirulina phycobiliprotein, it is difficult to separate the technical bottleneck of phycocyanin and allophycocyanin
[0003] At present, the separation and purification of spirulina phycocyanin and allophycocyanin usually use hydroxyapatite medium for chromatographic separation, but there are weak adsorption and elution time Long time, small amount of sample, etc., and it needs to be combined with other multi-step chromatography operations such as size exclusion chromatography and ion exchange chromatography to further improve the purity of phycocyanin
It has also been reported that the purity of phycobiliprotein can be improved by repeated salting-out operations or two-phase extraction technology, without the need for multi-step chromatography operations, but multi-step extraction operations also have problems such as low product yields and long operating cycles, especially for those with Phycocyanin with fluorescent properties, its fluorescent properties are easy to lose during the long-term extraction process

Method used

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  • Spirulina phycocyanin and extraction method thereof
  • Spirulina phycocyanin and extraction method thereof
  • Spirulina phycocyanin and extraction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 prepares phycocyanin extract

[0046] (1) Preparation of Spirulina powder suspension: Weigh 5g of Spirulina platensis powder and dissolve it in 100mL of phosphate buffer (0.01mol / L, pH=7) to prepare Spirulina powder suspension;

[0047] (2) Preparation of spirulina cell disruption solution: put the spirulina powder suspension at 4°C for 4 hours, then freeze at -20°C, mince the ice cubes, and repeat freezing and thawing at 4°C to -20°C. Broken until the operation was repeated three times, centrifuged at 9000rpm for 30min, and the spirulina cell broken liquid was obtained;

[0048] (3) Preparation of phycocyanin crude extract 1: First, add 17.6g of ammonium sulfate to the spirulina cell disruption solution, stir well, dissolve to 30% saturation, let stand at 4°C for 12 hours, centrifuge at 9000rpm for 30min to take the supernatant , then add 12.7g of ammonium sulfate to the supernatant, stir well, dissolve to 50% saturation, let it stand at 4°C for 12 hours,...

Embodiment 2

[0063] Example 2 Antioxidant Activity Determination Index Evaluation

[0064] The sample solution in this embodiment is the phycocyanin extract in different extraction steps.

[0065] DPPH system: take 2mL sample solution respectively, add 2mL of 1×10 -4 mol / LDPPH solution, after mixing, react in the dark at room temperature for 2 minutes, and centrifuge at 4000rpm for 10 minutes, and take the supernatant for testing. An equal volume of deionized water was used instead of the sample solution as the control group, and an equal volume of 95% ethanol was used instead of the DPPH solution as the blank group. Measure the absorbance A at 517nm, and extract the solvent (phosphate buffer saline or PEG20000 / Na 2 SO 4 The lower phase solvent of the two-phase system) and 95% ethanol mixed solution are used as a blank to adjust to zero, and six groups of parallel experiments are done, and the average value is taken.

[0066] Clearance rate (%)=[1-(A sample group-A blank group) / A contr...

Embodiment 3

[0073] Embodiment 3 Fluorescent Activity Measurement Index Evaluation

[0074] Fluorescence microscope observation method: DEAE-SephadexA50 pretreatment: Weigh 5g of DEAE-SephadexA50 and suspend it in 500mL distilled water, soak for 1h, pour off the upper layer of fine particles, add 0.5M NaOH at a ratio of 1(g):15(mL) and soak for 30min, pump Filtered, washed with distilled water to pH = 7, and then repeated the above process with 0.1M HCl. Take 5mL of 0.1mg / mL separated and purified phycocyanin product solution (0.01mol / LpH=7.0 phosphate buffer as solvent), add 100mg of pretreated and filtered DEAE-SephadexA50 adsorption medium, and separate at 4°C After static adsorption for 4h and 12h, the UV-visible absorbance values ​​A620 and A652 in the solution were measured with a spectrophotometer respectively, and the phycocyanin adsorbed by 1g DEAE-SephadexA50 adsorption medium was calculated to be 7.91mg and 15.76mg respectively. The DEAE-SephadexA50 adsorbed phycocyanin was was...

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Abstract

The invention relates to an extraction method of spirulina phycocyanin. The method comprises the following steps of: processing spirulina powder suspension by adopting a freeze thaw method and a machine crushing process to obtain spirulina cell disruption liquid; performing fractional salting out to precipitate the spirulina cell disruption liquid with 30 percent ammonium sulfate liquid and 50 percent ammonium sulfate liquid to obtain phycocyanin coarse extract liquid 1; adding PEG 20000 (polyethylene glycol 20000) and NaCl to further precipitate out phycocyanin coarse extract with higher purity; dissolving the phycocyanin coarse extract with phosphate buffer to prepare phycocyanin coarse extract liquid 2; and extracting the phycocyanin coarse extract liquid with PEG 20000 / Na2SO4 aqueous two-water-phase system, and desalting extracted lower phase by dialysis to obtain phycocyanin fine extract liquid, and lyophilizing to prepare a phycocyanin finished product. The method is simple in extraction process and low in cost, and is suitable for intermittent and scale production processing of high-purity and high-yield spirulina phycocyanin finished product. The phycocyanin coarse extract can be stored for a long time, and the phycocyanin finished product which is prepared by further purification has excellent antioxidant free radical scavenging effect and fluorescent strength.

Description

technical field [0001] The invention relates to the technical field of deep processing of natural products, in particular to a spirulina phycocyanin and an extraction method thereof. Background technique [0002] Spirulina contains a variety of biologically active substances, among which phycobiliprotein is rich in spirulina dry powder (accounting for 15-25, w / w%), and has important application values ​​in medicine, health care, and bioengineering. For the strains of Spirulina platensis commonly cultivated in the spirulina industry in my country, the phycobiliprotein components mainly include phycocyanin and allophycocyanin, and the content ratio of the two in phycobiliprotein is about 3:1, of which Due to the absolute advantage of phycocyanin content (accounting for about 10-20, w / w% of Spirulina dry powder), its research application is more common. Phycocyanin not only has the functions of anti-radiation, anti-cancer, immune regulation, promoting hematopoiesis and delaying...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/795C07K1/30
Inventor 刘杨虞永蕾钟名其肖湘赵永杰
Owner 丽江美之源食品有限公司
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