Novel protein glycosylation grafting method
A technology of protein glycosylation and glycosylated protein, which is applied in the field of protein glycosylation grafting to achieve the effect of overcoming uneven reaction and strong industrial operability
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Embodiment 1
[0036] Accurately weigh 0.5g soybean protein and 1.5g dextran, dissolve in 10mL 0.01mol / L sodium phosphate buffer solution with pH 6.5, stir at room temperature for 2h, add 3 drops of NaN 3 (0.02% w / w) for anti-corrosion, placed at 5°C for 24h, kept at a constant temperature of 60°C, and stirred for 30h at a stirring speed of 500r / min. After the reaction is completed, the reaction is rapidly cooled to room temperature to terminate the reaction, and freeze-dried to obtain a glycosylated protein product.
Embodiment 2
[0046]Put 1g of PEG2000 as a crowding reagent in the reaction device, add 10mL of 0.01mol / L sodium phosphate buffer solution with pH 7.0, stir for 2 hours to make it fully hydrated and dissolved, add 0.2g of soybean protein and 0.6g of dextran to the crowding In the solution of the reagent, after fully stirring for 2 hours, add NaN 3 (0.02% w / w) anti-corrosion, placed at 5°C for 24h, stirred at 50°C for 48h, cooled rapidly to 24°C after the reaction to obtain glycosylated protein product, purified by gel filtration chromatography to remove unreacted protein and glucose Polysaccharides and inert polymer PEG2000.
Embodiment 3
[0055] Accurately weigh 0.6g soybean protein and 3.6g dextran, dissolve in 20mL 0.01mol / L sodium phosphate buffer solution with pH 7.5, stir at room temperature for 2h, add 3 drops of NaN 3 (0.02% w / w) for anti-corrosion, placed at 5°C for 24h, kept at a constant temperature of 70°C, and stirred at a stirring speed of 500r / min for 12h. After the reaction was completed, it was rapidly cooled to room temperature to terminate the reaction, and freeze-dried.
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