Preparation method for micro-molecular glycopeptide of jellyfish
A small-molecule sugar and jellyfish technology, which is applied in the field of preparation of jellyfish small-molecule glycopeptides, can solve the problems of unfavorable glycopeptide structure research and large molecular weight of glycopeptides.
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Embodiment 1
[0019] One, the preparation of jellyfish glycoprotein, its operation is as follows:
[0020] 1) Grind the fresh jellyfish with a tissue masher, weigh 200g of the tissue mash solution, add 800ml of NaCl solution with a concentration of 0.5mol / L, mix well, and extract in a constant temperature shaker at 10°C for 10h. Then take it out and send it to a centrifuge, centrifuge for 20min under the condition of a centrifugal speed of 4000r / min, get the supernatant, and set aside;
[0021] 2) Concentrate the supernatant in step 1 to 1 / 20 of the original volume with a hollow cellulose membrane, add ethanol with a mass concentration of 95% to the concentrated supernatant while stirring, when the mass concentration of ethanol in the solution is When it reaches 75%, stop adding ethanol, let it stand for 2 hours, centrifuge for 15 minutes under the condition of centrifugal speed of 10000r / min, take the precipitate and freeze-dry it to be crude glycoprotein, and set aside;
[0022] 3) Use D...
Embodiment 2
[0029] One, the preparation method of jellyfish glycoprotein is as embodiment 1;
[0030] 2. The preparation of jellyfish small molecule glycopeptides, the operation steps are as follows: Take 1 g of glycoprotein separated and purified by ion exchange and gel column chromatography and dissolve it in distilled water to make a glycoprotein solution with a mass concentration of 0.1 mg / mL. Add compound protease to the protein solution for enzymolysis, the amount of compound protease added is 0.5% of the mass of the glycoprotein solution, enzymolysis at pH 8.0 and temperature 65°C for 2 hours to obtain glycopeptide solution, using ultrafiltration method, The obtained glycopeptide solution was graded, ultrafiltered with 3000Da, and the filtrate was concentrated and freeze-dried to obtain jellyfish small molecule glycopeptides. It was determined that 90% of the glycopeptides had molecular weights less than 3000Da.
Embodiment 3
[0032] One, the preparation method of jellyfish glycoprotein is as embodiment 1;
[0033] 2. The preparation of jellyfish small molecule glycopeptides, the operation steps are as follows: Take 1 g of glycoprotein separated and purified by ion exchange and gel column chromatography and dissolve it in distilled water to make a glycoprotein solution with a mass concentration of 0.1 mg / mL. Add compound protease to the protein solution for enzymolysis, the amount of compound protease added is 0.7% of the mass of the glycoprotein solution, enzymolysis is carried out at pH 8.0 and temperature 65°C for 2 hours to obtain a glycopeptide solution, and ultrafiltration is used. The obtained glycopeptide solution was graded, ultrafiltered with 3000Da, and the filtrate was concentrated and freeze-dried to obtain jellyfish small molecule glycopeptides. It was determined that 90% of the glycopeptides had molecular weights less than 3000Da.
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