Lepista sordida protein LsAPII as well as preparation method and application thereof

A technology of flower face mushroom and protein, which is applied in the preparation method of peptides, botanical equipment and methods, applications, etc., can solve the problems of no clear active substances, etc., and achieve the effects of good inhibition effect, simple preparation method, and easy-to-obtain raw materials

Inactive Publication Date: 2012-05-23
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 "Apricot mushroom extract and its preparation method and application" (application number: 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 there is no specific active substance, so further research and exploration are needed

Method used

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  • Lepista sordida protein LsAPII as well as preparation method and application thereof
  • Lepista sordida protein LsAPII as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 The preparation of the protein LsAPII of the flower face mushroom

[0038] Proceed as follows:

[0039] (1) Add 2 g of the fruiting body powder of Amanita chinensis into 20 ml of 0.025 M, pH 8.0 phosphate buffer, soak for 2 hours, centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0040] (2) Add ammonium sulfate to the supernatant obtained in step (1) until the saturation is 50%, place it at 4° C. for 8 hours, centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0041] (3) Add ammonium sulfate to the supernatant obtained in step (2) until the saturation is 70%, place it at 4° C. for 8 hours, centrifuge at 10,000 rpm for 25 minutes, and collect the precipitate.

[0042] (4), dissolve the precipitate obtained in step (3) with 0.025M, pH8.0 phosphate buffer solution, then dialyze through 3 times the weight of double distilled water for 8 hours, and use the dialysate obtained by d...

Embodiment 2

[0044] Example 2 The preparation of the protein LsAPII of the flower face mushroom

[0045] Proceed as follows:

[0046] (1) Add 2 g of the fruiting body powder of Pleurotus chinensis into 30 ml of 0.03 mol / L, pH 9.0 phosphate buffer, soak for 3 hours, centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0047] (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.

[0048](3) Add ammonium sulfate to the supernatant obtained in step (2) until the saturation is 70%, place it at 6° C. for 12 hours, centrifuge at 10,000 rpm for 25 minutes, discard the supernatant, and collect the precipitate.

[0049] (4) Dissolve the precipitate obtained in step (3) with 0.03M, pH9.0 phosphate buffer solution, then dialyze with 1 times the weight of double distilled water for 16 hours, and use ...

Embodiment 3

[0051] Example 3 The preparation of the protein LsAPII of the flower face mushroom

[0052] Proceed as follows:

[0053] (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.

[0054] (2) Add ammonium sulfate to the supernatant obtained in step (1) until the saturation is 50%, place it at 2° C. for 6 hours, centrifuge at 10,000 rpm for 25 minutes, discard the precipitate, and take the supernatant.

[0055] (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.

[0056] (4) Dissolve the precipitate obtained in step (3) with 0.01mol / L, pH 6.0 phosphate buffer solution, then dialyze with 2 times the weight of double distilled water for 13 hours, and use AmiconUltra-15 3KD and A 30KD u...

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Abstract

The invention discloses a lepista sordida protein LsAPII, wherein the N-terminal amino acid sequence is DGEYVNDLEGTISVDA. The preparation method comprises the following steps: soaking lepista sordida sporophore powder in phosphate buffer solution, and centrifugating to take supernatant; then adding ammonium sulfate to a saturation of 50 percent and a saturation of 70 percent respectively and standing at a temperature of 0-6 DEG C for 6 to 16 hours; respectively centrifugating to take supernatant or precipitation; reusing phosphate buffer solution to dissolve, dialyze, ultra-filtrate by an ultra-filtration pipe, separate through an anion exchange column, and elute by 0.3 mol / L NaCl, and then obtaining lepista sordida protein LsAPII. The LsAPII has good inhibitory effect on tobacco mosaic virus, and then the LsAPII is identified as single protein, which provides the basis for the genetic engineering applications. As the LsAPII is a natural extract, the invention is harmless to humans and environments. In addition, the preparation method of the LsAPII is simple and has the advantages of easily-obtained raw material and lower production cost.

Description

technical field [0001] The invention belongs to the field of bio-pesticides, and in particular relates to a protein LsAPII of Azalea chinensis and a preparation method and application of the protein. Background technique [0002] Plant virus disease is second only to fungal disease, which causes serious losses to agricultural production every year. For example, the global annual loss caused by tobacco mosaic virus alone can reach 100 million US dollars. At present, there is no effective treatment for tobacco mosaic virus disease. Prevention methods. [0003] Edible fungi 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K1/36C07K1/34C07K1/30C07K1/18A01N37/46A01P1/00
Inventor 燕继晔李兴红周莹严红马凤金
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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