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Lepista sordida protein LsAPI, preparation method and application thereof

The technology of the mushroom and protein is applied to the preparation methods of peptides, botanical equipment and methods, applications and other directions, can solve the problem of no clear active substances and the like, and achieve the effects of good inhibition effect, easy availability of raw materials and simple preparation method.

Inactive Publication Date: 2010-03-17
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent application "Flower mushroom extract and its preparation method and application" (200810223439.5) discloses a flower face mushroom extract and flower face mushroom protein extract, and their application in the prevention and treatment of tobacco mosaic virus disease, but it is not clear Specific active substances, therefore, are subject to further research and exploration

Method used

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  • Lepista sordida protein LsAPI, preparation method and application thereof
  • Lepista sordida protein LsAPI, preparation method and application thereof
  • Lepista sordida protein LsAPI, preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0039] The preparation of embodiment 1 flower face mushroom protein LsAPI

[0040] Prepare as follows:

[0041] (1) Add 2 g of the fruiting body powder of Pleurotus chinensis into 20 ml of 0.025 M, pH 8.0 phosphate buffer and soak for 2 hours, then centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0042] (2) Add ammonium sulfate to the supernatant obtained in step (1) until the saturation is 50%, place it at 4° C. for 8 hours, then centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0043] (3) Ammonium sulfate was added to the supernatant obtained in step (2) until the saturation was 70%. After standing at 4°C for 8 hours, centrifuge at 10,000 rpm for 25 minutes to collect the precipitate.

[0044] (4) The precipitate after centrifugation was dissolved with 0.025mol / L, pH8.0 phosphate buffer solution, and then dialyzed with 3 times the weight of double distilled water for 10 hours, and the d...

Embodiment 2

[0046] The preparation of embodiment 2 flower face mushroom protein LsAPI

[0047] Prepare as follows:

[0048] (1) Add 2 g of the fruiting body powder of Pleurotus chinensis into 30 ml of 0.03 M, pH 9.0 phosphate buffer and soak for 3 hours, then centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0049](2) Add ammonium sulfate to the supernatant obtained in step (1) until the saturation is 50%, place it at 6° C. for 12 hours, centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0050] (3) Add ammonium sulfate to the supernatant obtained in step (2) until the saturation is 70%. After 12 hours at 6° C., centrifuge at 10,000 rpm for 25 minutes, discard the supernatant, and collect the precipitate.

[0051] (4) Dissolve the precipitate obtained in step (3) with 0.03mol / L, pH9.0 phosphate buffer solution, then dialyze with 1 times the weight of double distilled water for 13 hours, and use Amicon U...

Embodiment 3

[0053] Example 3: Preparation of the protein LsAPI from the flower face mushroom

[0054] Prepare as follows:

[0055] (1) Add 2 g of the fruiting body powder of Amanita variegata into 50 ml of 0.01 mol / L, pH 6.0 phosphate buffer, soak for 1 hour, centrifuge at 10,000 rpm for 25 minutes, and take the supernatant.

[0056] (2) Add ammonium sulfate to the supernatant obtained in step (1) until the saturation is 50%, place it at 2° C. for 6 hours, then centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0057] (3) Add ammonium sulfate to the supernatant obtained in step (2) until the saturation is 70%, place it at 2° C. for 6 h, centrifuge at 10,000 rpm for 25 min, discard the supernatant, and collect the precipitate.

[0058] (4) Dissolve the precipitate obtained in step (4) with 0.01M, pH6.0 phosphate buffer solution, then dialyze through 2 times the weight of double distilled water for 16 hours, and use the dialysate obtained by dialy...

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Abstract

The invention discloses a lepista sordida protein LsAPI and a preparation method thereof. The preparation method of the LsAPI comprises the steps: adding lepista sordida entity powder into phosphate buffer solution to be soaked and centrifuged; adding ammonium sulfate into supernate until degree of saturation being 50%-70%, placing for 6-16h at the temperature of 0-6 DEG C, and centrifuging; dissolving precipitation by the phosphate buffer solution, dialyzing by re-distilled water, and carrying out ultra-filtration by an ultra-filtration tube to obtain protein solution; and separating by an anion exchange column, eluting by 0.02mol / L of Tris-HCl with the pH value of 8.0, and obtaining the LsAPI which is the substance contained in the eluent. The LsAPI is a single protein and provides the foundation for genetic engineering application thereof; secondly, the LsAPI has good virus inhibition for tobacco mosaic virus, and is a natural extract so as to be harmless to both the human being andthe environment; and furthermore, the preparation method of the LsAPI is easy, the raw materials are easily obtained, and the cost is lower.

Description

technical field [0001] The invention belongs to biopesticides, and in particular relates to the protein LsAPI of the flower face mushroom and its preparation method and application. Background technique [0002] Plant virus disease is second only to fungal disease, which causes serious losses to agricultural production every year. For example, the annual loss caused by tobacco mosaic virus (TMV) in the world can reach 100 million U.S. dollars. There is no effective control method. [0003] Edible mushrooms contain a variety of proteins, which are not only rich in nutrition, but also contain many active substances, which have certain effects on improving human immunity, preventing cancer, and fighting cancer. At present, the research on antiviral active substances in edible fungi mainly focuses on the anti-animal virus, and there are few researches on the anti-plant virus. [0004] Lepista sordida belongs to Basidiomycotina, Hymenomycetes, Agaricales, Tricholomataceae, Lepi...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/36C07K1/34C07K1/30C07K1/18A01N47/44A01P1/00
Inventor 李兴红继晔燕严红周莹马凤金
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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