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Method for rapidly preparing glycopeptides

A glycopeptide, rapid technology, applied in the field of polypeptide glycosylation grafting, to achieve the effect of overcoming uneven reaction, strong industrial operability, and improving efficiency

Inactive Publication Date: 2012-01-11
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the advantages of convenient operation, short reaction time, and superior functionality of the obtained glycopeptides. It overcomes the shortcomings of the existing protein glycosylation graft modification methods that are difficult to realize industrialization, and the product has a large-scale application prospect in the food industry.

Method used

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  • Method for rapidly preparing glycopeptides
  • Method for rapidly preparing glycopeptides
  • Method for rapidly preparing glycopeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Prepare soybean protein into a solution with a mass fraction of 10%, heat it to 37°C, add pepsin (enzyme: protein amount = 1:100), enzymolyze it for 30 minutes, heat it to 100°C for 3 minutes to inactivate the enzyme, centrifuge (5000rpm, 20min), take the supernatant and freeze-dry it, which is soybean polypeptide. Put 1.0g dextran as a crowding reagent in a closed container, add 10mL 0.01mol / L sodium phosphate buffer solution with pH 6.5, stir for 2h to fully dissolve it, add 0.5g soybean polypeptide and 1.0g dextran to the In the solution containing the crowding reagent, after fully stirring for 2 hours, add 2 drops of NaN 3 (0.02% w / w) anti-corrosion, after standing at 5°C overnight, stirring at 60°C for 4h, cooling to 25°C to terminate the reaction after the reaction, centrifuging (8000rpm, 20min), and freeze-drying the supernatant to obtain glycopeptide products.

[0028] The above-mentioned glycopeptide products were used to prepare emulsions, and after standing ...

Embodiment 2

[0036]Prepare soybean protein into a solution with a mass fraction of 10%, heat it to 37°C, add pepsin (enzyme: protein amount = 1:100), enzymolyze it for 30min, then heat it to 100°C for 3min to inactivate the enzyme, centrifuge (5000rpm, 20min), take the supernatant and freeze-dry it, which is soybean polypeptide. Put 2.0g dextran in the reaction device as a crowding reagent, add 10mL 0.01mol / L sodium phosphate buffer solution with pH 5.5, stir for 2h to fully dissolve it, add 0.25g soybean polypeptide and 0.25g dextran to the reaction device In the solution containing the crowding reagent, after fully stirring for 2 hours, add 2 drops of NaN 3 (0.02% w / w) anti-corrosion, after standing overnight at 5°C, stir at a constant temperature of 70°C for 4 hours, after the end of the reaction, quickly cool to 25°C to terminate the reaction, centrifuge (8000rpm, 20min), and freeze-dry the supernatant to obtain the glycopeptide product .

[0037] The above-mentioned glycopeptide pro...

Embodiment 3

[0045] Prepare defatted soybean powder into a solution with a mass fraction of 10%, heat it to 55°C, add neutral protease (enzyme: protein amount = 1:100), enzymolyze it for 30min, then heat it to 100°C for 3min to inactivate the enzyme, centrifuge ( 5000rpm, 20min), take the supernatant and freeze-dry it, which is soybean polypeptide. Put 1.0g of polyethylene glycol 2000 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 fully dissolve it, and mix 1.0g of soybean polypeptide and 0.5g of dextran Add to the solution containing the crowding reagent, stir well for 2h, add 2 drops of NaN 3 (0.02% w / w) anti-corrosion, after standing at 5°C overnight, stir at constant temperature 50°C for 6h, after the reaction is completed, quickly cool to 25°C to terminate the reaction, centrifuge (8000rpm, 20min), and freeze-dry the supernatant to obtain the glycopeptide product .

[0046] The above-mentioned gly...

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Abstract

The invention discloses a method for rapidly preparing glycopeptides. The method comprises the following steps: 1, carrying out enzymolysis on a protein to prepare polypeptide, deactivating an enzyme when the enzymolysis reaction is finished, centrifuging, taking a supernatant, and drying; 2, putting a dissolving reagent in a reaction apparatus, adding a buffer solution, and fully dissolving; 3, adding polysaccharides and polypeptides obtained in step 1 adding to a solution obtained in step 2, fully stirring for 2h, adding with 0.02%w / w NaN3 for anticorrosion, allowing to stand for a night at 5DEG C, stirring for 1-8h at 50-70DEG C, rapidly cooling to a temperature of below 25DEG C after a reaction is finished, and removing the dissolving reagent if the dissolving reagent is saccharosan 70 or polyethylene glycol 2000; and 4, centrifuging, taking a supernatant, and drying to obtain products of glycopeptides. The method has the advantages of convenient operation, short reaction time and high reaction efficiency, and the products glycopeptides obtained with the method which have superior functionality have wide application prospects in the food industry.

Description

technical field [0001] The invention belongs to the field of food, and relates to protein modification and a method for preparing glycopeptides, in particular to a method for polypeptide glycosylation and grafting. Background technique [0002] Most natural proteins have polysaccharide side chains linked by covalent bonds. Glycoproteins are widely found in various forms in animals, plants and some microorganisms. They play a key role in organisms, such as cell signal recognition, Various physiological functions such as growth regulation, intercellular information transmission, and immune regulation. Therefore, glycoproteins have attracted widespread attention. However, it is very difficult to separate and purify glycoproteins, and the preparation by genetic engineering technology is also very limited. Therefore, in biochemical, immune cell biology and medical research, the partial structure of glycoproteins—glycopeptides must be chemically synthesized as model molecules for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23J3/34C08B37/00
Inventor 齐军茹卓秀英杨晓泉
Owner SOUTH CHINA UNIV OF TECH
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