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Preparation method for pea aspartic acid protease inhibitor

A technology of protease inhibitors and aspartic acid, which is applied in the direction of protease inhibitors, peptide sources, animal/human peptides, etc., can solve the problems of environmental pollution, waste of protein resources, etc., achieve wide application, easy material acquisition, and prevent waste of resources Effect

Inactive Publication Date: 2013-02-27
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 50 g of peas and bake them 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 carry out leaching at 4 °C for 2 h. Stir every hour. After the extraction, the extract was centrifuged at 8000 r / min and 4°C for 30 min to obtain 175 mL of supernatant. The supernatant was water bathed at 60°C for 20 min, cooled to room temperature, 8000 r / min , Centrifuge at 4°C for 30 min to obtain 160 mL of supernatant, the concentration ratio of protein to sugar was measured to be about 0.98, the pepsin inhibitory unit was about 0.97 IU / mL, and the recovery rate was 87.6%; the supernatant was subjected to ionization For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, 30 cm × 2.2 cm, and the equilibration buffer was 10 mmol / L sodium phosphate buffer (pH 6.8). Elution mode: After 2 column volumes of equilibration buffer were eluted, 0.2, 0.4, 0.6, an...

Embodiment 2

[0031] Wash 50 g of peas and bake them 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 carry out leaching at 4 °C for 2 h. Stir every hour. After the extraction, the extract was centrifuged at 8000 r / min and 4°C for 30 min to obtain 175 mL of supernatant. The supernatant was water bathed at 60°C for 20 min, cooled to room temperature, 8000 r / min , Centrifuge at 4°C for 30 min to obtain 160 mL of supernatant, the concentration ratio of protein to sugar was measured to be about 1.05, the pepsin inhibitory unit was about 0.99 IU / mL, and the recovery rate was 88.4%; the supernatant was subjected to ionization For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, 30 cm × 2.2 cm, and the equilibration buffer was 10 mmol / L sodium phosphate buffer (pH 6.8). Elution mode: After 2 column volumes of equilibration buffer were eluted, 0.2, 0.4, 0.6, and ...

Embodiment 3

[0033] Wash 50g of peas and bake them 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 carry out leaching at 4°C for 2 hours, every half an hour. Stir once. After the extraction, the extract was centrifuged at 8000 r / min and 4°C for 30 min to obtain 175 mL of supernatant. The supernatant was water bathed at 60°C for 20 min, cooled to room temperature, 8000 r / min , Centrifuge at 4 °C for 30 min to obtain 160 mL of supernatant, the concentration ratio of protein to sugar was measured to be about 1.06, the pepsin inhibitory unit was about 0.98 IU / mL, and the recovery rate was 87.4%; the supernatant was subjected to ionization For exchange chromatography, 20 mL of crude inhibitor was taken each time for DEAE-52 ion exchange chromatography, 30 cm × 2.2 cm, and the equilibration buffer was 10 mmol / L sodium phosphate buffer (pH 6.8). Elution mode: After 2 column volumes of equilibration buffer were eluted, ...

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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 Patents(China)
IPC IPC(8): C07K14/81C07K1/18
Inventor 田亚平张秋萍
Owner JIANGNAN UNIV
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