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Preparation method for pea aspartic acid protease inhibitor

A protease inhibitor, aspartic acid technology, applied in the direction of protease inhibitors, peptide preparation methods, chemical instruments and methods, can solve the problems of protein resource waste, environmental pollution, etc. The effect of pollution

Inactive Publication Date: 2012-04-11
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vermicelli mainly uses the starch in the pea, and the protein in the pea is discharged along with the sewage, which not only pollutes the environment, but also causes a waste of protein resources

Method used

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  • Preparation method for pea aspartic acid protease inhibitor
  • Preparation method for pea aspartic acid protease inhibitor
  • Preparation method for pea aspartic acid protease inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Wash and bake 50 g of peas immediately, grind them into powder, add 200 mL of ultrapure water at a ratio of 1:4 (w / v), put them in the refrigerator, and extract them at 4°C for 2 h, during which time every half Stir every hour. After leaching, centrifuge the extract at 8000 r / min at 4°C for 30 min to obtain 175 mL of supernatant, put the supernatant in a water bath at 60°C for 20 min, cool to room temperature, and centrifuge at 8000 r / min , centrifuged at 4°C for 30 min to obtain 160 mL of supernatant, the measured concentration ratio of protein to sugar was about 0.98, the pepsin inhibitory unit was about 0.97 IU / mL, and the recovery rate was 87.6%; the supernatant was ionized For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, the size 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 vo...

Embodiment 2

[0031] Wash and bake 50 g of peas immediately, grind them into powder, add 200 mL of ultrapure water at a ratio of 1:4 (w / v), put them in the refrigerator, and extract them at 4°C for 2 h, during which time every half Stir every hour. After leaching, centrifuge the extract at 8000 r / min at 4°C for 30 min to obtain 175 mL of supernatant, put the supernatant in a water bath at 60°C for 20 min, cool to room temperature, and centrifuge at 8000 r / min , centrifuged at 4°C for 30 min to obtain 160 mL of supernatant, the measured concentration ratio of protein to sugar was about 1.05, the pepsin inhibitory unit was about 0.99 IU / mL, and the recovery rate was 88.4%; the supernatant was ionized For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, the size 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 volu...

Embodiment 3

[0033] Wash and bake 50g of peas immediately, grind them into powder, add 200 mL of ultrapure water at a ratio of 1:4 (w / v), put them in the refrigerator, and extract them at 4°C for 2 hours, during which every half hour Stir once. After leaching, centrifuge the extract at 8000 r / min at 4°C for 30 min to obtain 175 mL of supernatant, put the supernatant in a water bath at 60°C for 20 min, cool to room temperature, and centrifuge at 8000 r / min , centrifuged at 4°C for 30 min to obtain 160 mL of supernatant, the measured concentration ratio of protein to sugar was about 1.06, the pepsin inhibitory unit was about 0.98IU / mL, and the recovery rate was 87.4%; the supernatant was ionized For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, the size 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...

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Abstract

The invention discloses a preparation method for a pea aspartic acid protease inhibitor, and belongs to the technical field of a plant extraction biological activity product. The method comprises the following steps of: directly grinding pea into powder; leaching with water; performing high-speed centrifugation; thermally treating supernatant; performing high-speed centrifugation again; separating the purified aspartic acid protease inhibitor from the supernatant by a DEAE (Diethylaminoethyl)-52 ion exchange column chromatography; and then freeze-drying under vacuum to obtain the aspartic acid protease inhibitor dry powder. The aspartic acid protease inhibitor prepared by the invention has higher inhibition activity on pepsin and rennin in aspartic acid protease family, is directly extracted from pea, is low in cost and safe in application, and has broad application prospect in the fields of food additives, medicines, agriculture and the like.

Description

technical field [0001] A preparation method of a pea aspartic acid protease inhibitor, specifically, the peas are ground into powder, extracted with water, centrifuged at high speed, heat treated, and the supernatant is subjected to DEAE-52 ion exchange chromatography to obtain relatively pure aspartame The invention relates to an amino acid protease inhibitor product, which belongs to the technical field of plant extraction bioactive products. Background technique [0002] Biologically active substances refer to some foods that contain a variety of biologically active compounds, which can cause various biological effects after interacting with the body. There are many kinds of them, including sugars, lipids, protein polypeptides, sterols, alkaloids, glycosides and so on. They are mainly found in plant foods, and some are beneficial to people, while others are harmful. In the fields of disease prevention and treatment, food, etc., bioactive substances are still a topic wor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/81C07K1/18
Inventor 田亚平张秋萍
Owner JIANGNAN UNIV
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