Preparation method of antibacterial polypeptides from elaeagnus angustifolia seeds and application of antibacterial polypeptides

A technology of antibacterial peptides and seeds, which is applied to the preparation method of peptides, antibacterial drugs, antifungal agents, etc., and can solve the problem of excessive seed protein

Active Publication Date: 2016-10-12
XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] According to the results of domestic and foreign literature surveys on the chemical components and biological activities of the seeds of Elaeoptera, the current reports on the chemical constituents of the seeds mainly focus on polysaccharides, flavonoids and volatile oils. In addition, there are also related reports on protein, fat a

Method used

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  • Preparation method of antibacterial polypeptides from elaeagnus angustifolia seeds and application of antibacterial polypeptides
  • Preparation method of antibacterial polypeptides from elaeagnus angustifolia seeds and application of antibacterial polypeptides
  • Preparation method of antibacterial polypeptides from elaeagnus angustifolia seeds and application of antibacterial polypeptides

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Experimental program
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Effect test

Embodiment 1

[0015] a. Wash 300g of Elaeagnus radix fruit with water, clean and dry the seeds naturally, pulverize them, and degrease with petroleum ether to obtain 48g of degreasing Elaeoptera erae seeds;

[0016] b. With the defatted seeds of Elaeagnus eraeae obtained in step a, at room temperature, use 10% acetic acid with a concentration of 10% acetic acid to extract the total polypeptide of the seeds of Elaeoptera eraeaceae under stirring at a ratio of w / v=1:7 at a temperature of 20° C. for a period of time After 2 hours, filter the supernatant, then pour the supernatant into anhydrous acetone for overnight precipitation according to v / v=1:5, centrifuge at 4°C and 12000r / min for 3 times, each time for 15min, and Each precipitate was washed with cooled anhydrous acetone, and then the precipitate was naturally dried to obtain the total polypeptide extract. The total protein content was measured by the biuret method, and the total protein content was 72.6% by ultraviolet detection at 540n...

Embodiment 2

[0019] Use the agar plate method to detect the antibacterial effect of the sample on the three bacteria, and measure the size of the antibacterial ring of the sample on the three bacteria, so as to judge the antibacterial ability of the sample:

[0020] Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia coli were cultured overnight in LB liquid medium at 175rpm and 37°C until the OD600 was around 2.0 (1×106CFU / mL). Mix 10 μL of solution with 8 mL of LB medium containing 0.7% agar and spread it on the LB medium containing 25 mL of 1.5% agar. , cultivate overnight at 37°C, and measure the diameter of the inhibition zone. Ampicillin was used as a positive control, and normal saline was used as a negative control.

[0021] surface 1 Antibacterial results of antibacterial peptides from seeds of Eleuthero jujube (M±m)

[0022]

Embodiment 3

[0024] The tandem mass spectrometer is composed of two mass spectrometers connected in series. The first mass analyzer pre-separates or adds energy modification to the ions, and the second mass analyzer analyzes the results; Mass spectrometry analysis was performed on the seed bold polypeptide, and four polypeptide molecules with molecular weights of 5492.9Da, 6551.5Da, 4400.5Da and 2837Da were obtained by mass spectrometry analysis.

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Abstract

The invention relates to a preparation method of antibacterial polypeptides from elaeagnus angustifolia seeds and application of antibacterial polypeptides. The preparation method comprises the following steps: treating dry elaeagnus angustifolia seeds through grinding and degreasing, performing acetic acid extraction and acetone precipitation, performing reverse-phase silica gel column chromatography (C18), separating and purifying to obtain an antibacterial part of the elaeagnus angustifolia seeds. Polyacrylamide gel electrophoresis analysis on the obtained antibacterial part of the elaeagnus angustifolia seeds shows that the antibacterial part is a mixture of polypeptide with the molecular weight of 4400Da-14000Da and protein, and the antibacterial part is verified to have the effect of effectively inhibiting activities of candida albicans, escherichia coli and staphylococcus aureus. The antibacterial polypeptides can be applied as raw materials or auxiliary agents to preparation of health-care products or other functional food.

Description

technical field [0001] The present invention relates to the preparation method and application of the antibacterial polypeptide of sandy jujube seeds Background technique [0002] Elaeagnus is a deciduous tree or small tree of the genus Elaeagnus (Elaeagnus), which can be eaten directly and is a favorite fruit of the Uighur people in Xinjiang. It is recorded in the "Chinese Pharmacopoeia Uyghur Subvolume" that the main pharmacological activities include anti-oxidation, anti-proliferation, anti-hemolysis, anti-allergic, antibacterial, anti-viral, etc., which have high research and medicinal value. Elaeoptera also has the functions of invigorating the spleen and stomach, calming the nerves, calming down, and stopping diarrhea and astringent intestines. In addition to being used for feeding, Elaeoptera is also a good tree species for afforestation, greening, firewood, windbreak and sand fixation. Elaeagnus as feed has a long history in Northwest China. Its leaves and fruit a...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/20A61K38/16A61P31/10A61P31/04A23L33/18
CPCA23V2002/00A61K38/00C07K14/415A23V2200/10A23V2200/30A23V2250/55Y02A50/30
Inventor 阿布力米提·伊力阿吉艾克拜尔·艾萨艾合米丁·外力肖开提·萨力霍
Owner XINJIANG TECHN INST OF PHYSICS & CHEM CHINESE ACAD OF SCI
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