Method for extracting phycobiliprotein from spirulina

Abstract

The invention provides a method for extracting phycobiliprotein from spirulina. The method comprises the following steps: adding water into spirulina mud to obtain a spirulina cell suspension, performing ultrasonic treatment to obtain spirulina lysate, centrifuging the lysate and taking the lysate, sequentially filtering through filter membranes which are respectively 0.8mu m and 0.45mu m, to obtain phycocyanin crude extracting liquid, then sequentially ultrafiltrating the crude extracting liquid through an ultrafiltration membranes with molecular weight cut-off of 300KD and 100KD, collecting a filtrate of the molecular section, and performing freezing and drying to the filtrate to obtain phycobiliprotein powder. According to the method, the defect of using chemical reagent to extract phycobiliprotein in the prior art is overcome, no residuals exist in the preparation, and the environment pollution does not exist. The extraction rate of the phycobiliprotein is greater than 80%, the content of phycobiliprotein is more than 60%, and the purity of the phycobiliprotein is more than 1.5. The overall technological process is safe and reliable, fast and efficient. Compared with the traditional method, by using the method, the content of the phycobiliprotein is high before using of chromatography, the phycobiliprotein with the purity not only can meet the requirement of food and cosmetic industries, but also can be applied to aspects of medical and health products, and the like.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to a method for extracting phycobiliprotein from spirulina. Background technique [0002] Spirulina contains a variety of biologically active substances, and its nutritional value has been widely used. As the main active pigment protein in spirulina, namely phycobiliprotein, it also contains phycocyanin, allophycocyanin and phycoerythrin. Phycobiliproteins can not only be widely used as natural pigments in food, cosmetics, dyes and other industries, but also because of their strong fluorescence, phycobiliproteins can be developed into fluorescent reagents for medical diagnosis, immunochemistry and biological engineering and other fields. At the same time, because phycobiliprotein has obvious biological activities such as anti-oxidation, anti-inflammation, improving immunity, promoting animal cell regeneration, and inhibiting cancer cells, phycobiliprotein can also be made into f...

AI Technical Summary

Problems solved by technology

[0003] At present, the extraction methods of phycobiliproteins in spirulina include many methods. Although fresh algae is used in the disclosed method of CN1268516A, the fresh algae must be dehydrated, and then the phycocyanin is proposed by freezing and thawing, and then filtered with a plate and frame, and finally used The ultrafiltration membrane with a molecular cutoff of 100KD removes small molecules with a molecular weight of less than 100,000 Daltons, and finally uses hydroxyapatite column chromatography to purify. The whole process is cumbersome and requires too many equipments, which is inconvenient for industrial production

The method disclosed in CN1027316545A is mainly to extract phycocyanin from spirulina powder by microfiltration and isoelectric point method, its structure is partially damaged, still contains many miscellaneous proteins, and its phycobiliprotein content is only about 30%. Moreover, the phycocyanin powder obtained due to the low content is green but not blue, and its application value is not high

Although the method disclosed in CN103613661A produces phycocyanin with a purity >4.0, the processing is relatively cumbersome, and the steps of spirulina pretreatment, chromatographic decolorization, hydroxyapatite purification, desalination, and concentration are required, and the process is complicated. Such phycocyanin purity Out of touch with practical applications, such high purity is not required for food and cosmetic applications; and the purity is too low for biological agents to be suitable for practical applications

In the disclosed method of CN102993297A, PEG20000 / Na 2 SO 4 Two-phase system extraction, the operation is relatively simple, but this method finally needs to dialyze and desalt phycocyanin, which increases the time cost. At the same time, more chemical reagents are used, the waste liquid treatment is more troublesome, and it is easy to cause secondary pollution to the environment.

In the disclosed method of CN102329381A, the extraction method of freezing and thawing plus ultrasonic crushing is adopted, ammonium sulfate salting out is used for purification, a large amount of chemical reagents are used in the extraction process, and the step process of gradient elution with hydroxyapatite column is relatively complicated, and the unit time The output is low, and it is difficult to realize industrialized mass production

The common defect of the above methods is that more or less chemical reagents are used, which will potentially pollute the environment. The extraction technology is relatively simple, and the obtained phycocyanin content is low. The technology of extracting relatively high content phycocyanin The operation is too complicated, and it is difficult to realize industrialized mass production