Method for efficiently extracting selenoprotein and application
A high-efficiency technology for selenoproteins, applied in the field of selenoprotein extraction, can solve the problems of inapplicable large-scale preparation of selenoproteins, weak selenium enrichment ability, and degradation of algal species, so as to increase the conversion rate of inorganic selenium and the content of selenoproteins , low cost and mature industrial chain
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Embodiment 1
[0043] The screening of embodiment 1 selenium-enriched spirulina mutant
[0044] Selenium is a trace element required for biological growth, so the selenium content of microalgae in the natural environment is often low. In addition, algae strains in nature will have problems such as algae degradation and production reduction during long-term cultivation. Therefore, it is very important to screen algal species with strong selenium-enrichment ability, high selenoprotein content and fast growth rate. In this embodiment of the present invention, artificial domestication and ultraviolet mutagenesis are combined to improve the selenium-enriching ability of Spirulina.
[0045] Wild Spirulina platensis (Spirulina platensis) FACHB-882 has poor tolerance to selenium and can only tolerate 360mg / L Na 2 SeO 3 . In this application, the method of artificial domestication is first adopted to improve the selenium-tolerant ability of Spirulina FACHB-882, and the 360mg / L Na 2 SeO 3 Cultur...
Embodiment 2
[0057] The groping of embodiment 2 selenoprotein extraction process
[0058] In this embodiment of the present invention, it is proposed to use an acid-base combination method to extract selenoproteins in spirulina. First, we compared the extraction efficiency of selenoproteins by the two methods of "alkali-soluble acid precipitation" and "acid-soluble alkali precipitation". The operation steps of "acid-dissolving and alkali-precipitating method" are as follows: dissolve 13.3g of broken algae powder in 200mL of 2% acetic acid solution; stir at 80 rpm for 2h; centrifuge at 6000rpm for 15min at 10°C to obtain precipitates A and Supernatant A; take supernatant A, add NaOH to adjust the pH to 10, let stand for 1 hour to fully precipitate the protein; centrifuge at 6000rpm, 15min, 10°C to obtain precipitate B and supernatant B; precipitate B is the obtained protein. The operation steps of "alkali-dissolved acid precipitation method" are as follows: dissolve 13.3g of broken algae po...
Embodiment 3
[0064] The decolorization of embodiment 3 selenoproteins
[0065] Algae cells contain chlorophyll and other pigments. These pigments will be mixed in the selenoprotein extracted by the above-mentioned "alkali-soluble acid precipitation" method, resulting in a greenish color of the selenoprotein, affecting the appearance and quality of the selenoprotein, and it needs to be decolorized. deal with. In the present invention, we use ethanol and acetone, these two commonly used decolorizers, to decolorize selenoproteins to remove impurities such as chlorophyll. Before decolorization, the selenoprotein of spirulina was green, and the protein was soft and muddy. Use 200mL of absolute ethanol and acetone to resuspend the protein, stir for 1 hour, and then centrifuge at 6000rpm for 10 minutes to decolorize the selenoprotein. Hardened ( Figure 5 ), indicating that both have good decolorization effects.
[0066] By observing the eluent of ethanol and acetone, we found that the color ...
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