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Preparation method of potato aspartic proteinase inhibitor

A protease inhibitor, aspartic acid technology, applied in the direction of protease inhibitors, peptide preparation methods, chemical instruments and methods, etc., can solve problems such as high pollution, achieve pollution elimination, prevent resource waste, and easy operation. Effect

Inactive Publication Date: 2009-12-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, potatoes are mostly used for starch production in addition to eating, and the protein liquid produced in the production is generally discharged as wastewater, which is very polluting.

Method used

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  • Preparation method of potato aspartic proteinase inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: After 682.7g potatoes are washed, peeled and cut into pieces, press 0.5g Na 2 SO 3 Add 0.341g Na / kg potato 2 SO 3 To prevent polyphenol oxidation, after homogenization, centrifuge at 8000r / min, 4°C for 30min to obtain 510mL supernatant, autoclave the supernatant at 121°C for 20min, cool to room temperature, centrifuge at 14000r / min, 4°C for 20mim, The precipitate was discarded, and the collected 432mL supernatant was concentrated with a 10KDa ultrafiltration membrane, and ultrafiltered at a pressure of 0.075MPa and room temperature for 10min to obtain a concentrated solution of 112mL, with an inhibitory rate of about 5.19U / mg to pepsin, protein and sugar The concentration ratio is about 1.47, and the yield is about 86.1%. Add 28mL-20°C ethanol to the concentrate, centrifuge at 10000r / min and 4°C for 10min, take the supernatant and add 160.5mL-20°C ethanol, 10000r / min , After centrifuging at 4°C for 10 minutes, take the precipitate; add 40 mL of pH 6.8 phos...

Embodiment 2

[0029] Embodiment 2: After 887.6g potatoes are washed, peeled and cut into pieces, press 0.5g Na 2 SO 3 Add 0.443g Na / kg potato 2 SO 3To prevent polyphenol oxidation, after homogenization, centrifuge at 8000r / min, 4°C for 30min to obtain 562mL supernatant, autoclave the supernatant at 121°C for 20min, cool to room temperature, centrifuge at 14000r / min, 4°C for 20mim, The precipitate was discarded, and the collected 480mL supernatant was concentrated with a 10KDa ultrafiltration membrane, and ultrafiltered at a pressure of 0.075MPa and room temperature for 10min to obtain a concentrated solution of 95mL with an inhibitory rate of about 4.65U / mg to pepsin. The concentration ratio is about 1.23, and the yield is about 84.6%. Add 23.75mL-20°C ethanol to the concentrate, centrifuge at 10000r / min and 4°C for 10min, take the supernatant and add 142.5mL-20°C ethanol, 10000r / min After centrifuging for 10 min at 4°C for 10 min, take the precipitate; add 40 mL of pH 6.8 phosphate buff...

Embodiment 3

[0030] Embodiment 3: After 507.3g potatoes are washed, peeled and cut into pieces, press 0.5g Na 2 SO 3 Add 0.254g Na / kg potato 2 SO 3 To prevent polyphenol oxidation, after homogenization, centrifuge at 8000r / min, 4°C for 30min to obtain 321mL supernatant, autoclave the supernatant at 121°C for 20min, cool to room temperature, centrifuge at 14000r / min, 4°C for 20mim, The precipitate was discarded, and the collected 250mL supernatant was concentrated with a 10KDa ultrafiltration membrane, and ultrafiltered at a pressure of 0.075MPa and room temperature for 10min to obtain a concentrated solution of 87mL, with a specific inhibitory activity to pepsin of about 3.98U / mg. The concentration ratio of sugar is about 1.36, and the yield is about 81.9%. Add 21.75mL-20°C ethanol to the concentrate, centrifuge at 10000r / min and 4°C for 10min, take the supernatant and add 130.5mL-20°C ethanol, 10000r After centrifugation at 10000r / min and 4°C for 10 minutes, take the precipitate; add 4...

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Abstract

The invention relates to a preparation method of potato aspartic proteinase inhibitor and belongs to the technical field of bioactive products extracted from plant. The preparation method comprises the following steps: directly peeling, cutting and mixing potatoes evenly; carrying out high-speed centrifugation; completing high-pressure sterilization of the supernatant fluid and then carrying out high-speed centrifugation again; completing hyperfiltration and concentration; carrying out fractionated precipitation of ethanol; eliminating undissolved substance through centrifugation after deposition and redissolution; and finally, obtaining the dry powder of the aspartic proteinase inhibitor through vacuum freeze drying. The aspartic proteinase inhibitor prepared by the preparation method has higher inhibitory activity on pepsin and can be directly extracted from potatoes, thereby having low cost and application safety without causing pigment influence; therefore, the inhibitor has broad application prospects in the fields of foodstuff, medicament, agriculture, and the like.

Description

technical field [0001] A method for preparing a potato aspartic acid protease inhibitor, specifically, using potato homogenate to obtain potato juice, high-pressure sterilization, high-speed centrifugation, ultrafiltration concentration, and ethanol fractional precipitation to obtain an aspartic acid protease inhibitor The product belongs to the technical field of plant extraction bioactive products. Background technique [0002] Protease inhibitors generally refer to a class of substances that have an inhibitory effect on proteases, participate in the control of proteolytic activities in organisms, and are of great significance to maintaining normal life activities. It has a wide range of sources and exists in the tissues and organs of various animals and plants. According to the different active centers of the target proteases it inhibits, it can be divided into the following types: serine protease inhibitors; cysteine ​​protease inhibitors; aspartic protease inhibitors; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/81C07K1/36C07K1/34C07K1/30
Inventor 田亚平孙莹
Owner JIANGNAN UNIV
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