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Preparation of soybean anti-fatigue biological active peptide

A bioactive peptide and anti-fatigue technology, applied in the field of plant protein extraction and processing, can solve the problems of low anti-fatigue activity, high cost, and low purity

Inactive Publication Date: 2013-10-09
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology for preparing anti-fatigue active peptides has problems such as high cost, low anti-fatigue activity and low purity of the product

Method used

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  • Preparation of soybean anti-fatigue biological active peptide
  • Preparation of soybean anti-fatigue biological active peptide
  • Preparation of soybean anti-fatigue biological active peptide

Examples

Experimental program
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Effect test

preparation example Construction

[0013] A preparation of soybean anti-fatigue bioactive peptide, using highly denatured soybean meal as raw material, adding water to prepare a mixed solution with a substrate concentration of 6-10%, and performing ultrasonic pretreatment on the mixed solution, the ultrasonic power is 300-500W, The ultrasonic time is 10-30min, and the ultrasonic temperature is 45-55°C; neutral protease is added to the treated mixed solution for enzymolysis to obtain an enzymolysis solution. The enzymolysis time is 3-5h, and the amount of enzyme added is 28000-36000U / g substrate; after enzymolysis, the enzymolysis solution was subjected to ultrafiltration treatment by membrane separation technology to obtain bioactive peptides with high peptide content and molecular weight less than 5 kDa, which were then separated and purified by anion exchange chromatography and reversed-phase high performance liquid chromatography. Finally, the soybean anti-fatigue bioactive peptide was obtained by freeze-dry...

Embodiment 1

[0018] Embodiment 1: the screening experiment of ultrasonic pretreatment optimal parameter

[0019] 1 Materials and methods

[0020] 1.1 Materials and reagents

[0021]

[0022] 1.2 Main instruments and equipment

[0023]

[0024] 1.3 Test method

[0025] 1.3.1 Process flow

[0026] Highly denatured soybean meal→add water to make a solution→ultrasonic pretreatment→enzymolysis→inactivate enzyme→cooling→adjust pH to acidity→centrifuge→supernatant→freeze-drying→antioxidant active peptide

[0027] 1.3.2 Determination of the free radical scavenging rate of diphenylpicryl (DPPH)

[0028] Take 1.00 mL of the hydrolyzate, add 4.00 mL of 100 μmol L-1 DPPH ethanol solution and mix well, keep it in the dark for 30 min, adjust to zero with the original solvent, measure the absorbance at 517 nm and record it as A i ; Add 1.00 mL of solvent to 4.00 mL of 100 μmol L-1 DPPH solution in the same way and mix well, measure the absorbance as A c ; Take 1.00 mL of hydrolyzate and add 4...

Embodiment 2

[0048] Embodiment 2: the screening test of optimal parameter of enzymatic hydrolysis process

[0049] 1 Materials and methods

[0050] 1.1 Materials and reagents

[0051]

[0052] 1.2 Main instruments and equipment

[0053]

[0054] 1.3 Test method

[0055] 1.3.1 Process flow

[0056] Highly denatured soybean meal→add water to make a solution→ultrasonic pretreatment→enzymolysis→inactivate enzyme→cooling→adjust pH to acidity→centrifuge→supernatant→freeze-drying→antioxidant active peptide

[0057] 1.3.2 Determination of the free radical scavenging rate of diphenylpicryl (DPPH)

[0058] Take 1.00 mL of the hydrolyzate, add 4.00 mL of 100 μmol L-1 DPPH ethanol solution and mix well, keep it in the dark for 30 min, adjust to zero with the original solvent, measure the absorbance at 517 nm and record it as A i ; Add 1.00 mL of solvent to 4.00 mL of 100 μmol L-1 DPPH solution in the same way and mix well, measure the absorbance as A c ; Take 1.00 mL of hydrolyzate and add...

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Abstract

The invention discloses preparation of a soybean anti-fatigue biological active peptide, belonging to an extraction processing technology of plant protein. The method comprises the following steps: adding water in high-denaturation soybean meal serving as a raw material to prepare a mixed solution; performing ultrasonic preprocessing on the mixed solution; adding neutral protease into the processed mixed solution to perform enzymolysis so as to obtain enzymatic hydrolysate; performing ultrafiltration treatment on the enzymatic hydrolysate by using a membrane separation technology after the enzymolysis to obtain biological active peptide with high peptide content and molecular weight of less than 5 kDa; then performing separation and purification by using anion exchange chromatography and reversed-phase high-performance liquid chromatography; and finally performing freeze drying to obtain the soybean anti-fatigue biological active peptide. The soybean anti-fatigue biological active peptide prepared by the preparation method disclosed by the invention has higher autioxidant enzyme activities and lower malondialdehyde (MDA) content after movement of a mouse; and results show that the soybean peptide has the functions of reducing cell injury caused by lipid oxidation to have an anti-fatigue effect by removing free radicals produced in the body and inhibiting the generation of lipid peroxides.

Description

technical field [0001] The invention belongs to the extraction and processing technology of plant protein, and mainly relates to the preparation of a soybean anti-fatigue bioactive peptide. Background technique [0002] Soybean meal after pressing or extracting contains about 45% protein. The content of amino acids in soybean protein is relatively balanced. Except for the slightly low content of methionine, the content of other amino acids necessary for the human body is relatively high. At present, soybean meal is mainly used for feed, which greatly wastes soybean protein resources, so it is necessary to make full use of soybean protein in soybean meal through biotechnology and other methods. Soybean peptide is a mixture of oligopeptides with a relative molecular mass below 5 000 obtained by enzymatic hydrolysis of soybean protein. Its amino acid composition is basically the same as that of soybean protein, and its essential amino acid content is rich. Soybean peptide has...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/18C07K1/20A61P39/00
Inventor 许晶江连洲赵青山李杨齐宝坤冯红霞曹亮王欢
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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