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Antiphytoviral black pleurotus ostreatus protein as well as preparation method and application thereof

A technology of oyster mushroom and protein is applied in the field of anti-plant virus black oyster mushroom protein and its preparation, which can solve the problem of lack of clear anti-plant virus effect and the like, and achieve the effects of good inhibition effect, simple preparation method and low cost.

Active Publication Date: 2012-01-04
中农绿康(北京)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patent application "A Black Oyster Mushroom Extract and Its Preparation Method and Application" (Application No.: 200810223438.0) discloses a black oyster mushroom protein extract with anti-plant virus effect, but it is not clear which of the protein extracts The active substance has anti-plant virus effect, which needs further research and exploration

Method used

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  • Antiphytoviral black pleurotus ostreatus protein as well as preparation method and application thereof
  • Antiphytoviral black pleurotus ostreatus protein as well as preparation method and application thereof
  • Antiphytoviral black pleurotus ostreatus protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 The preparation of black oyster mushroom protein of the present invention

[0044] Proceed as follows:

[0045] (1) Mix 2 g of black oyster mushroom mycelium dry powder (purchased from Yichang Senyuan Edible Fungi Co., Ltd.) with 20 mL of phosphate buffer solution with a pH value of 8.0, extract at 4°C for 2 hours, and centrifuge at 10,000 rpm for 25 min; discard the precipitate , the supernatant is the crude extract of black oyster mushroom mycelium.

[0046] (2) Ammonium sulfate was added to the crude extract of black Pleurotus ostreatus mycelia to a saturation of 40%, placed at 4° C. for 8 hours, then centrifuged at 10,000 rpm for 25 minutes, and the supernatant was collected.

[0047] (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 10000rpm for 25min, collect the precipitate to obtain the crude extract of black oyster mushroom protein;

[0048] (4) Dissolving the ...

Embodiment 2

[0051] Embodiment 2 The preparation of black oyster mushroom protein of the present invention

[0052] Proceed as follows:

[0053] (1) Mix 2 g of black oyster mushroom mycelium dry powder with 20 ml of phosphate buffer solution with a pH value of 7.0, extract at 4° C. for 2 hours, centrifuge at 10,000 rpm for 25 minutes, and the supernatant is the rough extraction of black oyster mushroom mycelia;

[0054] (2) Ammonium sulfate was added to the crude extract of black oyster mushroom mycelia to a saturation of 40%, placed at 4°C for 8 hours, centrifuged at 10000rpm for 25min, and the supernatant was collected;

[0055] (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 10000rpm for 25min, collect the precipitate to obtain the crude extract of black oyster mushroom protein;

[0056] (4) Dissolving the obtained black oyster mushroom protein crude extract with 0.01mol / L, pH8.0 PBS buffer solutio...

Embodiment 3

[0059] Embodiment 3 Preparation of black oyster mushroom protein of the present invention

[0060] Extract using the following method:

[0061] (1) According to the mixing ratio of the quality of black oyster mushroom mycelia dry powder and the phosphate buffer solution whose pH value is 9.0 is 1g: 20mL, leach for 2 hours at 4°C, centrifuge at 10000rpm for 25min, take the supernatant to get black oyster mushroom crude mycelia extract;

[0062] (2) Ammonium sulfate was added to the mycelia crude extract of the obtained black oyster mushroom until the saturation was 40%, placed at 4°C for 6 hours, then centrifuged at 10000rpm for 25min, and the supernatant was taken;

[0063] (3) Add ammonium sulfate to the supernatant obtained in step (2) until the saturation is 70%, place it at 4° C. for 6 hours, then centrifuge at 10,000 rpm for 25 minutes, collect the precipitate, and obtain the crude extract of black oyster mushroom protein;

[0064] (4) Dissolving the obtained black oyst...

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Abstract

The invention discloses a black pleurotus ostreatus protein PoAP and a preparation method thereof. The preparation method comprises the following steps: firstly, lixiviating black pleurotus ostreatus hypha dry powder in phosphate buffer solution for 1-3 hours and then centrifugating to collect supernatant liquor; then adding ammonium sulfate to the collected supernatant liquor to a saturation of 40 percent and a saturation of 70 percent respectively, standing at 2-6 DEG C for 6-12 hours; and centrifugating to collect precipitation; using phosphate buffer solution to dissolve precipitation andthen utilizing distilled water to dialyze; then separating through an anion exchange column, and eluting by 0.5 m NaCl and collecting elution peak; and then ultra-filtrating at the speed of 5,000 rpmfor 40 minutes through a 30KD ultra-filtration pipe, then obtaining the black pleurotus ostreatus protein which is the material in the obtained solution. The black pleurotus ostreatus protein PoAP issingle protein which can be applied on genetic engineering; and the invention has good inhibitory effect on tobacco mosaic virus. As the black pleurotus ostreatus protein PoAP is natural extract, theinvention is harmless to humans and environments. In addition, the preparation method of the black pleurotus ostreatus protein 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 biopesticides, and in particular relates to a plant virus-resistant black oyster mushroom protein, a preparation method and application thereof. Background technique [0002] Plant virus diseases are various, widely distributed, and difficult to control, which cause serious losses to agricultural production every year (Du Chunmei et al., 2004), and are the second largest class of plant diseases after fungi. Tobacco mosaic virus (TMV) is a model species of Tobamovirus Group, which can occur from the seedbed to the field, and the incidence rate in the field is generally 5-20%, and the infection loss in the seedling stage or field stage Up to 30 to 50%, severe cases never harvest. [0003] At present, many types of chemical pesticides have been used in the prevention and treatment of plant virus diseases, but their effects are not ideal. With people's emphasis on the ecological environment, the development and applic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K1/36C07K1/34C07K1/30C07K1/18A01N47/44A01P1/00
Inventor 刘建华严红燕继晔周莹李兴红朱琳
Owner 中农绿康(北京)生物技术有限公司