Novel method for extracting high-purity R-phycoerythrin in laver
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A phycoerythrin and high-purity technology, which is applied in the preparation method of peptides, chemical instruments and methods, algae/bryopeptides, etc., can solve the problems of complex operation, solvent consumption, and many steps, and achieve high purity and yield. The effect of economical separation process and mild separation conditions
Pending Publication Date: 2021-12-07
HARBIN INST OF TECH AT WEIHAI
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Problems solved by technology
These traditional methods have too many steps, complex operation, solvent consumption, high kinetic energy consumption, and limited purity of the extracted protein, so additional multi-step chromatography is required to achieve the purity target, which is extremely unfavorable for industrial large-scale production of these proteins
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Embodiment 1
[0029] (1) Take 5 g of dried laver, add 250 mL of sterilized water and freeze overnight at -20 °C, after centrifugation, take the supernatant for salting out, and add ammoniumsulfate to 35% saturation.
[0032] (4) Establishment of a two-phase aqueous system: Add 300-400 μL of K with a pH of 8.0 to 0.30-0.50 g of a deep eutectic solvent 2 HPO 4 Salt solution, placed in a 20-25 ℃ water bath, that is, a two-phase aqueous system.
[0033] (5) Optimization of phycoerythrin extraction conditions: Add 300-400 μL of K with pH 8.0 to 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55 g DES 2 ...
Embodiment 2
[0037] (1) Take 5 g of dried laver, add 250 mL of sterilized water, and freeze overnight at -20 °C. After centrifugation, take the supernatant for salting out, and add ammonium sulfate to 35% saturation.
[0038] (2) Take the precipitate after centrifugation, and dialyze it in the phosphatebuffer solution with pH=6.8 to remove the ammonium sulfate, and obtain the crude phycoerythrin extract.
[0039] (3) Synthesis of deep eutectic solvent: take cholinechloride and urea as raw materials, stir at 80-90 °C for 2-3 hours, and obtain a clear and transparent liquid, that is, deep eutectic solvent.
[0040] (4) Establishment of a two-phase aqueous system: Add 300-400 μL of K with a pH of 8.0 to 0.30-0.50 g of a deep eutectic solvent 2 HPO 4 Salt solution, placed in a 25 ℃ water bath, that is, a two-phase aqueous system.
[0041] (5) Optimization of phycoerythrin extraction conditions
[0042] Add 300-400 μL of K pH 8.0 to 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55 g DES 2 HPO 4S...
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Abstract
The invention discloses a method for extracting high-purity phycoerythrin. The method comprises the following steps: repeatedly freezing and thawing laver to break cells, centrifuging, salting out and dialyzing to obtain supernatant crude protein; and using a novel green eutecticevaporate solvent, adding phosphate to form an aqueous two-phase system, and obtaining high-purity phycoerythrin through extraction. The phycoerythrin extracted by the method is high in purity, the applied eutecticevaporate solvent aqueous two-phase system is green, environment-friendly and good in biocompatibility, and the extraction method is low in cost and easy to scale and industrialize and has application prospects in the aspects of extracting the high-purity phycoerythrin, protecting the environment and the like.
Description
technical field [0001] The invention belongs to the technical field of protein extraction and purification, and in particular relates to a method for preparing high-purity phycoerythrin from laver by a deep eutectic solvent-two-phase method. Background technique [0002] Phycoerythrin (PE) is a kind of phycobiliprotein, which is isolated and purified from red algae. Phycoerythrin has good light-absorbing properties and high quantum yield. In addition, it can be easily combined with biotin and various monoclonal antibodies, so its wide application can be widely used as a natural pigment in food, In industries such as cosmetics and dyes, it can also be made into fluorescent reagents for clinical medical diagnosis, immunochemistry and bioengineering. It has important theoretical and practical significance for the extraction and research of phycoerythrin. [0003] At present, the separation and purification of phycoerythrin mainly include ammoniumsulfateprecipitation combine...
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