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A kind of mulberry leaf immunoactive peptide and its preparation method

An immunoactive peptide and an immunoactive technology, applied in the field of mulberry leaf immunoactive peptide and its preparation, can solve the problem that there is no research on the preparation of mulberry leaf immunoactive peptide, and achieve the improvement of the dual-use value of medicine and food, the mild purification process, and the expansion of the Application-wide effects

Active Publication Date: 2021-02-19
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are a few literatures and patents that analyze the extraction and optimization of mulberry leaf protein and mulberry leaf peptide, but there is no research on the preparation of mulberry leaf immunoactive peptides

Method used

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  • A kind of mulberry leaf immunoactive peptide and its preparation method
  • A kind of mulberry leaf immunoactive peptide and its preparation method
  • A kind of mulberry leaf immunoactive peptide and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0034] A method for preparing mulberry leaf immune activity peptide (neutral protease enzymatic hydrolysis), comprising the following steps:

[0035] (1) Preparation of crude mulberry leaf immunoactive peptides: fresh mulberry leaves were washed and dried, crushed and sieved to obtain mulberry leaf powder; 30 g of mulberry leaf powder was mixed with 600 mL of distilled water, dissolved and ultrasonically crushed for 15 min. After extracting at room temperature for 2 h, centrifuge (8000 g, 4 °C, 20 min), and add solid ammonium sulfate to the supernatant to reach 65% saturation. The precipitate was collected by centrifugation (centrifugal force 8000 g, 4°C, 20 min), dialyzed and desalted, and freeze-dried to obtain the mulberry leaf protein. 5 g of mulberry leaf protein was dissolved in water to prepare a solution with a mass concentration of 1%, and heated at 95 °C for 5 min. After cooling, the pH was adjusted to 7.0, and 1% neutral protease (Neutrase, 0.8 L) was added, and af...

Embodiment 2

[0044] A method for preparing mulberry leaf immunoactive peptide (compound enzymatic hydrolysis with alkaline protease and neutral protease), comprising the following steps:

[0045] (1) Preparation of crude mulberry leaf immunoactive peptides: fresh mulberry leaves were washed and dried, crushed and sieved to obtain mulberry leaf powder; 30 g of mulberry leaf powder was mixed with 900 mL of distilled water, dissolved and ultrasonically crushed for 20 min. After extracting at room temperature for 2 h, centrifuge (8000 g, 4 °C, 20 min), and add solid ammonium sulfate to the supernatant to reach 65% saturation. The precipitate was collected by centrifugation (8000 g, 4°C, 20 min), dialyzed and desalted, and freeze-dried to obtain the mulberry leaf protein. 5 g of mulberry leaf protein was dissolved in water to prepare a solution with a mass concentration of 5%, and heated at 95 °C for 5 min. After cooling, the pH was adjusted to 7.2, and 2% neutral protease (Neutrase, 0.8 L) wa...

Embodiment 3

[0051] A method for preparing mulberry leaf immune activity peptide (compound protease, papain compound enzymolysis), comprising the following steps:

[0052](1) Preparation of crude mulberry leaf immune active peptide: fresh mulberry leaves were washed and dried, crushed and sieved to obtain mulberry leaf powder; 30 g of mulberry leaf powder was mixed with 500 mL of distilled water, dissolved, ultrasonically crushed for 15 min, and extracted at room temperature After 2 h, centrifuge (8000 g, 4°C, 20 min), and add solid ammonium sulfate to the supernatant to reach 65% saturation. Centrifuge at 8000 g for 20 min, take the precipitate, dialyze and freeze-dry to obtain the mulberry leaf protein. 5 g of mulberry leaf protein was dissolved in water to prepare a solution with a mass concentration of 4%, and heated at 95 °C for 5 min. After cooling, adjust the pH to 7.0, add 2% compound protease (Protamex, Novozymes, Denmark), enzymatically hydrolyze at 50°C for 2 h, then inactivate...

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Abstract

The invention belongs to the technical field of mulberry leaf comprehensive utilization, and provides a mulberry leaf immunity active peptide and a preparation method thereof. The method comprises the following steps of using mulberry leaves as raw materials; preparing mulberry leaf protein by water extraction salt precipitation or alkali extraction acid precipitation; then, preparing a coarse product of the mulberry leaf immunity active peptide through single enzymolysis or composite enzymolysis by six kinds of enzymes including neutral protease, alkaline protease, composite protease, papain, flavourzyme and trypsase; performing separation and purification on the coarse product through anion column chromatography, Sephadex G50, G-25 or G-15 gel column chromatography and semi-preparation RP-HPLC; obtaining the immunity peptide with high activity and high purity. The immunity peptide can promote the macrophage RAW264.7 to secrete NO, interleukin IL-6 and tumor necrosis factor TNF-alpha; the resistance capability of the body on the pathogen invasion is enhanced. Meanwhile, the immunity active peptide can enhance the RAW264.7 endocytose neutral red capability.

Description

technical field [0001] The invention relates to a mulberry leaf active peptide, in particular to a mulberry leaf immune active peptide and a preparation method thereof. Background technique [0002] Immunomodulation refers to the interaction between immune cells and immune molecules in the immune system, as well as between the immune system and other systems, so that the body's immune response is at the most appropriate level, thereby maintaining the physiological dynamic balance in the organism. Once the immune regulation is out of balance, it will cause adverse symptoms such as pathogenic microbial infection, immunodeficiency disease, tumor and hypersensitivity. Therefore, it is particularly important to find immune molecules with immune activity. Recent studies believe that improving the immune regulation activity of the body through natural active substances such as immune peptides and immune polysaccharides is the most promising method to replace antibiotic treatment. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K1/20C07K1/18C07K1/16
CPCC07K1/16C07K1/18C07K1/20C12P21/06
Inventor 吴晖孙崇臻赖富饶武文佳闵甜
Owner SOUTH CHINA UNIV OF TECH
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