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Preparation method for aspartic protease inhibitor in enzymatic hydrolysis products of soybean protein isolate

A technology for protease inhibitor and soybean protein isolate, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of unguaranteed safety and the like

Inactive Publication Date: 2012-03-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the currently reported aspartic protease inhibitors are synthesized by chemical methods, and their safety cannot be guaranteed. Therefore, research on inhibitors extracted from natural organisms will have greater practical application value

Method used

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  • Preparation method for aspartic protease inhibitor in enzymatic hydrolysis products of soybean protein isolate
  • Preparation method for aspartic protease inhibitor in enzymatic hydrolysis products of soybean protein isolate
  • Preparation method for aspartic protease inhibitor in enzymatic hydrolysis products of soybean protein isolate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Prepare 300 mL of 5% soybean protein isolate solution, ① Take 100 mL of the solution and add 40,000 U of papain, adjust the pH to 6.5, put it in a water bath at 55°C for 200 min, and measure its effect on pepsin every 20 min. ②Take 100 mL of the solution and add 40000 U of flavor protease, adjust the pH to 3.0, put it in a water bath at 37°C for 200 min, and test its inhibitory activity on pepsin every 20 min; ③Take 100 40,000 U of trypsin was added to the solution, the pH was adjusted to 7.5, and it was placed in a water bath at 37°C for 200 min, and its inhibitory activity on pepsin was measured every 20 min. The results showed that papain enzymatically hydrolyzed soybean protein isolate for 60 min, and had the greatest inhibitory activity on pepsin, with an inhibitory activity of 3.6 IU / mL; flavor protease enzymatically hydrolyzed soybean protein isolate for 60 min, had the greatest inhibitory activity on pepsin, and the inhibitory activity was 3.6 IU / mL; 1.5 IU / mL; ...

Embodiment 2

[0033]Prepare 100 mL of a 5% soybean protein isolate solution, add 40,000 U of papain, adjust the pH to 6.5, bathe in water at 55°C for 60 min, and centrifuge at 8,000 r / min and 4°C for 30 min to obtain the supernatant 98 mL, the supernatant was placed in a water bath at 70°C for 20 min, cooled to room temperature, and centrifuged at 8000 r / min at 4°C for 30 min to obtain 92 mL of supernatant, which had an inhibitory activity against pepsin of about 3.3 IU / mL, the supernatant was subjected to ion-exchange chromatography, and 20 mL of crude inhibitor was taken each time for DEAE-52 ion-exchange chromatography, which was 30 cm × 2.2 cm, and the equilibrium buffer was 10 mmol / L sodium phosphate buffer ( pH 6.8). Elution mode: After elution with equilibration buffer for 2 times the column volume, add 0.2, 0.4, 0.6, 0.8 mol / L NaCl phosphate buffer with different concentrations in sequence to equilibration buffer for stage elution, the flow rate is 1.5 mL / min, 280 nm wavelength U...

Embodiment 3

[0035] Prepare 150 mL of 5% soybean protein isolate solution, add 60,000 U of papain, adjust the pH to 6.5, bathe in water at 55°C for 60 min, and centrifuge at 8,000 r / min and 4°C for 30 min to obtain the supernatant 147 mL, the supernatant was placed in a water bath at 70 °C for 20 min, cooled to room temperature, and centrifuged at 8000 r / min at 4 °C for 30 min to obtain 138 mL of supernatant, which had an inhibitory activity against pepsin of about 3.4 IU / mL, the supernatant was subjected to ion-exchange chromatography, and 20 mL of crude inhibitor was taken each time for DEAE-52 ion-exchange chromatography, which was 30 cm × 2.2 cm, and the equilibrium buffer was 10 mmol / L sodium phosphate buffer ( pH 6.8). Elution mode: After elution with equilibration buffer for 2 times the column volume, add 0.2, 0.4, 0.6, 0.8 mol / L NaCl phosphate buffer with different concentrations in sequence to equilibration buffer for stage elution, the flow rate is 1.5 mL / min, 280 nm wavelengt...

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Abstract

The invention provides a preparation method for an aspartic protease inhibitor in enzymatic hydrolysis products of soybean protein isolate, which belongs to the field of separation and purification of biological products. The invention relates to enzymatic hydrolysis of soybean protein isolate with papain for obtainment of a crude product of the aspartic protease inhibitor. The aspartic protease inhibitor is prepared from the crude product by carrying out heat treatment and high speed centrifugation on an enzymatic hydrolysate and carrying out DEAE-52 ion exchange chromatography and separation on an obtained supernatant, and dry powder of the aspartic protease inhibitor is obtained by carrying out vacuum freeze drying on the obtained aspartic protease inhibitor. The aspartic protease inhibitor prepared in the invention has high inhibitory activity on pepsin and chymosin in the family of aspartic protease; the aspartic protease inhibitor extracted from the natural processing material soybean protein isolate which has undergone enzymatic hydrolysis has good specificity, good stability, safety in application and an application prospect in a wide variety of fields, e.g., medicines andhealth care, food additives, control of insect diseases, etc.

Description

technical field [0001] A method for preparing an aspartic protease inhibitor in a soybean protein isolate enzymatic hydrolyzate. Specifically, papain is used to enzymolyze soybean protein isolate under specific conditions. The enzymatic hydrolyzate is subjected to heat treatment, high-speed centrifugation, and DEAE-52 ion exchange. Chromatography, to obtain purified aspartic protease inhibitors, belongs to the technical field of separation and purification of biological products. Background technique [0002] Enzyme preparations have been widely used in various fields such as food, brewing, pharmaceuticals, organic acids, starch sugar, feed industry, textiles, leather, detergents and health products. The application fields are becoming more and more extensive. Food is closely related to people's lives. Therefore The application value of enzymes in the food industry is becoming more and more important, and it is currently greatly reflected in baking, dairy products, meat proc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/18
Inventor 田亚平张国强
Owner JIANGNAN UNIV
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