Sargassum thunbergii lectin and preparation method thereof

A technology of sargassum and lectin, which is applied in the preparation method of peptides, chemical instruments and methods, algae/bryopeptides, etc., can solve the problems of separation and purification that have not been reported, and achieve the effect of high activity

Inactive Publication Date: 2012-09-26
DALIAN OCEAN UNIV +1
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Problems solved by technology

Although brown algae is also a kind of seaweed, because conventional extracts contain polyphenols, it has been prejudiced that the agglutination of brown algae extracts is just a pseudoagglutination phenomenon of polyphenols agglutinating blood cells. The researches are mainly focused on red algae and green algae, while on the Phaeophyta Roundrophyta Sargassum genus Sargassum ( Sargassum thunbergii O'Kuntze ) Separation and purification of lectin has not been reported

Method used

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  • Sargassum thunbergii lectin and preparation method thereof
  • Sargassum thunbergii lectin and preparation method thereof
  • Sargassum thunbergii lectin and preparation method thereof

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Embodiment Construction

[0014] a. Immediately after the sargassum is harvested, wash it with filtered seawater, absorb the water with gauze or filter paper, freeze it in a -20°C refrigerator, freeze it in a freeze dryer, and grind it into powder. g / ml) with a ratio of 1:20, add 0.15mol.L -1 NaCl in PBS buffer, soak at 4°C for 16h, centrifuge at 5000r / min for 30min, and take the supernatant;

[0015] b. Under the condition of ice-water bath and stirring, add ammonium sulfate powder to the supernatant, so that the saturation of ammonium sulfate in the supernatant reaches 85%. After stirring and mixing, centrifuge at 5000r / min for 30min, collect the precipitate, and discard the supernatant ;

[0016] c. Dissolve the precipitate with saline and dialyze with saline to free of SO 4 2- Finally, a 1.6×20cm DEAE-52 cellulose ion-exchange chromatography column was used, with a pH value of 7.5, 0.01mol / L -1 Tris-HCl buffer equilibrated to A 280 -1 Tris-HCl buffer, the gradient eluent II of the storage bot...

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Abstract

The invention discloses a sargassum thunbergii lectin and a preparation method thereof, wherein the preparation method of the sargassum thunbergii lectin is characterized by selecting sargassum thunbergii as a raw material, carrying out lyophilization, pulverization, immersion, centrifugation, ammonium sulfate fractionation, DEAE-52 ion-exchange chromatography and SephadexG-200 gel filtration chromatography, thus producing the sargassum thunbergii lectin with a molecular weight of 17 kD. The sargassum thunbergii lectin of the invention can agglutinate a wide range of cells, has agglutinative functions for erythrocytes of rabbits, dogs, sheep, crucians, chickens and humans (A, B, AB), and the like, wherein minimum concentrations of agglutination reactions with rabbit and chicken erythrocytes are 4.4mg / ml and 0.55 mg / ml respectively, and shows hemagglutinin activity for rabbit erythrocytes even after heating treatment at 100 DEG C for 30 min (the activity decreases by 75%). A sugar inhibition experiment shows that the sargassum thunbergii lectin is only inhibited by glycoproteins as bovine thyroglobulin and gamma-globulin, and the minimum inhibition concentrations are 1.25 mg.mL<-1> and 2.5 mg.mL<-1> respectively.

Description

technical field [0001] The invention relates to a lectin and a preparation method thereof, in particular to a sargassum agglutinin which agglutinates cells extensively and a preparation method thereof. Background technique [0002] Lectin is a protein or glycoprotein of non-immune origin with sugar specificity that can promote cell aggregation. Lectins are widely distributed in nature. From viruses to humans, lectins have been isolated from almost all species, among which plant lectins were first discovered. In 1966, Boyd et al. discovered the presence of seaweed lectins in some tropical seaweeds. After that, Blunden and Rogers et al. tested the agglutinating activity of the extracts from seaweeds along the British coast, but found that only a few seaweed extracts had no effect on untreated seaweeds. The treated human red blood cells had a certain agglutination effect, and then Hori et al. screened the Vietnamese seaweed agglutinin. Hori reported that until 1994, people is...

Claims

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Application Information

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IPC IPC(8): C07K14/405C07K1/36C07K1/30C07K1/18C07K1/16
Inventor 李丹彤周晓茹
Owner DALIAN OCEAN UNIV
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