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Method of preparing peptide having trypsin inhibition activity by using complex enzyme enzymolysis oyster

A trypsin inhibition and compound enzyme technology, which is used in drug combination, fermentation, antitumor drugs, etc., can solve the problems of difficult extraction and limited content of natural trypsin inhibitor, and achieve the effect of high product activity

Inactive Publication Date: 2007-08-08
OCEAN UNIV OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no report of extracting protease inhibitors from protein hydrolyzate so far
The main problem at present is that the content of natural trypsin inhibitor is very limited, and it is difficult to extract

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0006] Add oyster meat to 2-4 times, preferably 3 times the weight of the meat in Tris-HCl buffer solution (0.05M, pH8.5) for beating, and the initial pH of the system is 8-9, preferably 8.5. Add flavor alkaline protease (Alcalase) according to the ratio of 550~650U / g meat, preferably 600U / g meat, stir at 45~55°C, preferably 50°C, and enzymatically hydrolyze for 1~2 hours, preferably 1 hour, then inactivate the enzyme at 100°C, cool to 40-50°C, preferably 45°C; use hydrochloric acid to adjust the pH to 5-6, preferably 5.5, according to 650-750U / g meat, preferably 700U / g Add bromelain to the proportion of meat, stir, and enzymatically hydrolyze at 45-55°C, preferably 50°C, for 1-3 hours, preferably 2 hours, then inactivate the enzyme at 100°C, cool to room temperature, 4000r / min or more After centrifugation at a rotating speed, the supernatant was separated and purified by 10k ultrafiltration membrane and Sephadex G-25 column chromatography, and then freeze-dried to obtain a pe...

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PUM

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Abstract

The invention relates to a method to make trypsin activity restraining peptide by compound enzyme multiple step directional enzymolyzing oysters. The feather is that: adding oyster meat into 2-4 times weight Tris-Hcl buffer solution and pulping; according to 550-650U / g meat ratio to add flavor alkali protease, whisking in constant temperature and enzymolyzing for 1-2 hours under the temperature between 45-55 degree centigrade, then putting out enzyme, and cooling; using Hcl to adjust the PH value to 5-6, adding bromelain according to 650-750U / g meat ratio, whisking, enzymolyzing for 1-2 hours under 45-55 degree centigrade, then putting out enzyme, cooing, centrifuging, gathering the supernatant filtering, separating and purifying, then freezing out. The product has high activity and the restrain rate to the activity of commodity trypsin could reach 67%.

Description

technical field [0001] The invention relates to a method for preparing peptides with trypsin inhibitory activity by stepwise and directed enzymatic hydrolysis of oysters with compound enzymes. Background technique [0002] Protease inhibitors are a class of substances with tumor suppressive activity. At present, bovine lung trypsin inhibitors and human urinary trypsin inhibitors have been developed as adjuvant drugs for tumors and are widely used in clinical practice. Some people abroad have isolated protease inhibitors from the liver, eggs, epidermal mucus and muscle of Pacific pleated squid, skipjack, puffer fish, prawn, white croaker, long snake mullet and other aquatic products. Products are mainly used in the prevention and treatment of diseases. However, there is no report on extracting protease inhibitors from protein hydrolyzate so far. The main problem at present is that the content of natural trypsin inhibitor is very limited, and it is difficult to extract. Co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00A61P35/00
Inventor 曾名湧崔海英刘尊英董士远
Owner OCEAN UNIV OF CHINA
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