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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 can be 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: 2006-01-25
OCEAN UNIV OF CHINA
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  • Description
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AI Technical Summary

Problems solved by technology

However, there has been no report of extracting protease inhibitors from protein hydrolyzate so far
The main pro

Method used

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Experimental program
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Embodiment Construction

[0006] Add 2 to 4 times, preferably 3 times the weight of the oyster meat to Tris-HCl buffer (0.05M, pH 8.5) for beating, and the initial pH of the system is 8 to 9, preferably 8.5. Add flavored alkaline protease (Alcalase) at a ratio of 550-650U / g meat, preferably 600U / g meat, stir at a constant temperature of 45-55°C, preferably 50°C, and enzymatically hydrolyze for 1-2 hours, preferably It is 1 hour, then the enzyme is inactivated at 100°C, cooled to 40-50°C, preferably 45°C; pH is adjusted to 5-6 with hydrochloric acid, preferably 5.5, according to 650-750U / g meat, preferably 700U / g Add bromelain to the proportion of meat, stir, enzymolysis at a constant temperature of 45-55 ℃, preferably 50 ℃ for 1-3 hours, preferably 2 hours, then inactivate the enzyme at 100 ℃, cool to room temperature, and the temperature is more than 4000r / min. Centrifuge at a rotating speed, take the supernatant, go through 10k ultrafiltration membrane and Sephadex G-25 column chromatography for sepa...

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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 a peptide with trypsin inhibitory activity by using a compound enzyme to hydrolyze oyster in a step-by-step direction. Background technique [0002] Protease inhibitors are a class of substances with tumor inhibitory activity. Currently, 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 their livers, eggs, epidermal mucus and muscles using aquatic products such as Pacific squid, bonito, puffer fish, prawns, white croaker, and long snake mullet, respectively. Products are mainly used in the prevention and treatment of diseases. However, so far, there is no report on the extraction of protease inhibitors from protein hydrolyzates. The main problem is that the content of natural trypsin inhibitor is very limited, and the extraction is difficu...

Claims

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Application Information

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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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