Agaricus bisporus active ingredient extract as well as preparation method and application thereof

Abstract

The invention relates to the technical field of biological extraction, and particularly provides an agaricus bisporus extract as well as a preparation method and application thereof. The preparation method of the agaricus bisporus extract comprises the following steps: slicing agaricus bisporus, adding deionized water, carrying out microwave heating, and adding an alkaline substance to adjust the pH value, so as to obtain a first mixture; performing ultrasonic treatment on the first mixture while adding an acidic material to obtain a second mixture, and performing centrifugal treatment and filtration treatment on the second mixture to obtain a first filtrate; adding an alkaline substance into the first filtrate, performing stirring treatment and dialysis treatment, and finally performing concentration and freeze-drying treatment to obtain the agaricus bisporus extract. According to the invention, wall breaking is carried out in an alkaline environment by utilizing a microwave heat effect, so that fat-soluble and water-soluble active ingredients in cells are released; ultrasonic treatment is performed under an acidic condition, so that efficient dissolution and structural modification of polysaccharide, small-molecule sterol and phenolic compounds are promoted, the extraction rate and the purity are both improved, and the problems of low efficiency and low purity of a traditional method are solved.

Description

Technical Field

[0001] The present invention relates to the technical field of biological extraction, and in particular to an Agaricus bisporus active ingredient extract, a preparation method and an application thereof. Background Art

[0002] Obesity has become a global health concern, sparking extensive research into natural, safe, and effective fat-reducing and fat-blocking products. Agaricus bisporus, a common edible fungus, contains a variety of bioactive components with potential health benefits.

[0003] However, existing methods for preparing Agaricus bisporus extracts mostly use traditional processes such as water extraction or alcohol extraction, which often have problems such as low extraction efficiency, incomplete extraction of bioactive ingredients, many impurities in the extract, and insufficient purity and content of the target active components, limiting its in-depth development and industrial application in fat reduction and fat inhibition.

[0004] Therefore, there is an urgent need to provide a mild, controllable method for preparing an Agaricus bisporus extract that is rich in highly active ingredients. Summary of the Invention

[0005] The present invention provides an extract of active ingredients of Agaricus bisporus, as well as a preparation method and application. The extract utilizes microwave thermal effect to initially break the cell wall in an alkaline environment, thereby partially releasing the fat-soluble and water-soluble active ingredients in the cells. Subsequently, the extract is ultrasonically treated under acidic conditions to further promote the efficient dissolution and structural modification of polysaccharides, small molecular sterols and phenolic compounds through the cavitation effect, thereby enhancing their biological activity. At the same time, unlike the prior art of extracting with alkali or acid solution alone, the present invention uses acidic ultrasound as a secondary extraction method, thereby achieving a dual improvement in extraction rate and purity. Centrifugal filtration, secondary alkaline precipitation and dialysis are used to remove proteins, low molecular weight impurities and ineffective components, thereby improving the purity of the extract. Finally, the product is obtained by reduced pressure concentration and freeze-drying, thereby overcoming the problems of low efficiency and low purity of traditional methods, achieving efficient enrichment and retention of fat-blocking and fat-reducing active ingredients in Agaricus bisporus, and improving preparation efficiency and product quality.

[0006] The present invention provides a method for preparing an Agaricus bisporus extract, comprising the following steps: S100, adding Agaricus bisporus slices to deionized water, heating the mixture with a microwave, and adding an alkaline substance to adjust the pH value to obtain a first mixture; S200, ultrasonically treating the first mixture while adding an acidic substance to obtain a second mixture; S300, centrifuging and filtering the second mixture to obtain a first filtrate; S400, adding an alkaline substance to the first filtrate, stirring the mixture, dialyzing the mixture, and finally concentrating and freeze-drying the mixture to obtain the Agaricus bisporus extract.

[0007] In any of the above technical solutions, step S200 specifically includes: cooling the first mixture, performing ultrasonic treatment at 300-1000W for 2-8h, and simultaneously adding an acidic substance dropwise to control the pH value of the first mixture to be 2.5-5.5, to obtain a second mixture.

[0008] In any of the above technical solutions, the acidic substance includes at least one of hydrochloric acid and acetic acid.

[0009] In any of the above technical solutions, in step S100, the mass ratio of Agaricus bisporus slices to deionized water is 1:(5-10); the alkaline substance includes at least one of sodium hydroxide and potassium hydroxide; the pH value of the first mixture is 8-14; the microwave heating time is 4-8 hours, and the temperature is 90-180°C.

[0010] In any of the above technical solutions, the temperature of microwave heating is gradient controlled, including a first step temperature and a second step temperature; wherein the temperature of the first step temperature is between 90 and 120°C, and the temperature of the second step temperature is between 120 and 180°C.

[0011] In any of the above technical solutions, in step S300, the rotation speed of the centrifugal treatment is 3000-5000 r / min, and the time is 10-20 min.

[0012] In any of the above technical solutions, step S400 specifically includes: S410, adding an alkaline substance solution dropwise to the first filtrate, adjusting the pH value to 6-8, stirring for 30-60 minutes, and standing for 1-2 hours to obtain a precipitate; S420, dialyzing the precipitate in a dialysis bag with a molecular weight cutoff of 1000-5000 Da for 12-24 hours, and replacing the dialysis water 3-5 times during the dialysis treatment to obtain a dialyzed solution; S430, concentrating the dialyzed solution under reduced pressure to reduce the volume of the dialyzed solution to 1 / (5-10) of the original volume, and freeze-drying to obtain an Agaricus bisporus extract.

[0013] In any of the above technical solutions, in step S410, the second alkaline substance includes at least one of sodium hydroxide and potassium hydroxide; in step S430, the vacuum degree in the decompression condition is -0.06 to -0.08 MPa, and the temperature is 40 to 60°C. The present invention provides an Agaricus bisporus extract, which is prepared by any one of the above preparation methods.

[0014] The present invention also provides an application of an Agaricus bisporus extract, and the application of the Agaricus bisporus extract prepared by any of the above preparation methods in the preparation of foods or health products that inhibit fat absorption.

[0015] After adopting the technical solution of the present invention, the technical effects that can be achieved are as follows: 1. The rapid and uniform heating generated by microwave radiation during the microwave alkali addition stage causes the water molecules in the Agaricus bisporus cells to vibrate violently, causing the cell walls to rupture rapidly. The alkaline environment further disintegrates the polysaccharide and protein networks, making it easier for fat-soluble and water-soluble active ingredients to be released. 2. Acidic ultrasonic treatment uses the strong shear force and shock waves generated by ultrasonic cavitation to help dissolve the active substances remaining in the remaining cell fragments into the solution, destroy the protein-polysaccharide cross-links, and promote the hydrolysis of bound components. Compared with using acid extraction or alkaline extraction alone, the combined action of these two steps can obtain a higher yield of target active molecules in a shorter time, thereby greatly improving the efficiency of the entire extraction process; 3. After centrifugation and filtration, most insoluble impurities such as cell debris, non-polar oils, and cell membrane debris are effectively removed. A secondary alkaline treatment separates and precipitates some proteins, small molecular weight polyphenols, and non-target active polysaccharides from the active ingredients, further removing interfering substances. Dialysis treatment then completely removes salts, small molecular weight sugars, free amino acids, and some low-molecular weight degradation products, thereby making the active ingredients more concentrated and more stable, and reducing the risk of potential allergic and toxic side effects during oral administration or in vitro experiments. 4. The Agaricus bisporus extract prepared by the present invention has significant promoting and inhibiting effects on fat decomposition and absorption at different concentrations, and shows good fat-reducing and fat-blocking potential compared with control drugs on the market. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings to be used in describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. Those skilled in the art can also derive other drawings based on these drawings without inventive effort. Figure 1 This is a comparison of the fluorescence intensity of zebrafish yolk sac fat after treatment with samples from Example 1, Example 2, and a normal control group; Figure 2 1 is a comparison of the staining intensity of fat in the intestine and tail blood vessels of zebrafish after treatment with samples from Example 1, Example 2 and the normal control group. DETAILED DESCRIPTION

[0017] In order to more clearly understand the above-mentioned objects, features and advantages of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be noted that, in the absence of conflict, the embodiments of the present application and the features therein can be combined with each other.

[0018] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention may also be implemented in other ways different from those described herein. Therefore, the scope of protection of the present invention is not limited to the specific embodiments disclosed below.

[0019] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below.

[0020] Obesity has become a major global public health issue. Long-term high-fat diets and lack of exercise lead to fat accumulation, chronic low-grade inflammation, and various metabolic disorders, such as type 2 diabetes and cardiovascular disease, posing a serious threat to human health. Therefore, developing safe, effective, and naturally sourced fat-blocking and fat-reducing products has become a research priority in the food and pharmaceutical industries.

[0021] Agaricus bisporus is an edible fungus rich in bioactive compounds such as polysaccharides, sterols, and phenols. Studies have shown that its polysaccharide components can inhibit α-glucosidase activity, and that sterols and phenols can affect lipid metabolism pathways, potentially possessing physiological functions such as lipid inhibition, fat reduction, and hypolipidemic properties. Currently, most research and industrial processes use hot water extraction, alcohol extraction, acid-base immersion, or enzymatic hydrolysis to obtain Agaricus bisporus extracts.

[0022] However, these traditional methods have the following major shortcomings in practical applications: low extraction efficiency, low purity of active ingredients, significant ingredient loss, and poor fat-blocking / fat-reducing effects. Therefore, there is an urgent need to provide a mild, controllable method for preparing Agaricus bisporus extract that is rich in highly active ingredients.

[0023] In view of this, the present embodiment provides an extract of active ingredients of Agaricus bisporus, as well as a preparation method and application. The microwave thermal effect is used to initially break the cell wall in an alkaline environment, so that the fat-soluble and water-soluble active ingredients in the cells are partially released; then ultrasonic treatment is performed under acidic conditions, and the cavitation effect is further used to promote the efficient dissolution and structural modification of polysaccharides, small molecular sterols and phenolic compounds, thereby enhancing their biological activity; at the same time, unlike the existing technology of separate alkaline or acid extraction, the present invention uses acidic ultrasound as a secondary extraction method to achieve a double increase in extraction rate and purity. Centrifugal filtration, secondary alkaline precipitation and dialysis are used to remove proteins, low molecular impurities and invalid components to improve the purity of the extract; finally, the product is obtained by reduced pressure concentration and freeze-drying, thereby overcoming the problems of low efficiency and low purity of traditional methods, achieving efficient enrichment and retention of fat-blocking and fat-reducing active ingredients in Agaricus bisporus, and improving preparation efficiency and product quality.

[0024] Specifically, a method for preparing an Agaricus bisporus extract comprises the following steps: S100, adding Agaricus bisporus slices to deionized water, heating with a microwave, and adding an alkaline substance to adjust the pH value to obtain a first mixture; S200, adding an acidic substance while subjecting the first mixture to ultrasonic treatment to obtain a second mixture; S300, centrifuging and filtering the second mixture to obtain a first filtrate; S400, adding an alkaline substance to the first filtrate, and performing stirring treatment, dialyzing treatment, and finally concentrating and freeze-drying treatment to obtain an Agaricus bisporus extract.

[0025] Preferably, in step S100, the cell walls of Agaricus bisporus are destroyed and the active ingredients therein are dissolved by microwave addition of alkali, while the potential fat-blocking and fat-reducing active ingredients in the extract are fully released. By controlling the mass ratio of Agaricus bisporus to deionized water to 1: (5-10), adjusting the pH to 8-14 with alkaline substances, microwave heating for 4-8 hours, and controlling the temperature at 90-180°C, it is ensured that the cell walls are fully broken, excessive hydrolysis and thermal degradation are avoided, and the extraction efficiency and active ingredient content are improved; in addition, these parameter ranges have good repeatability, which is conducive to process scale-up and quality control, and ensures that the content of active ingredients and biological activity of each batch of extracts remain stable.

[0026] Furthermore, microwave heating adopts temperature gradient control, first using a relatively low temperature stage at 90-120°C to slowly destroy the cell structure and avoid thermal degradation of active ingredients due to excessive temperature; then entering a higher temperature stage at 120-180°C, the enhanced thermal effect accelerates the dissolution and structural transformation of more active substances, thereby ensuring the integrity of the active ingredients while achieving efficient extraction; step control avoids damage to the target active molecules caused by local overheating or rapid temperature rise during microwave heating, thereby improving the purity and biological activity of the extracted products.

[0027] Preferably, in step S200, the acidic environment can promote the hydrolysis and dissolution of certain bound active ingredients in Agaricus bisporus cells, making it easier for bioactive molecules such as polysaccharides and polyphenols to be released from the cell matrix; at the same time, the acidic condition is conducive to breaking the protein-polysaccharide cross-linking bonds in the cell wall structure, thereby increasing the concentration of the target active ingredients in the extract; in addition, by controlling the pH between 2.5 and 5.5, it can not only inhibit microbial growth and reduce impurity contamination, but also enhance the polarity and water solubility of certain fat-reducing or fat-blocking active ingredients at the chemical modification level, thereby improving their availability in subsequent analysis or bioactivity testing.

[0028] Furthermore, in terms of synergy with ultrasonic treatment, the strong micro-area shear force and shock wave effect generated by ultrasonic cavitation can further break up the cell structure in an acidic environment, allowing acidic hydrolysis to proceed more thoroughly. The combination of the two enables the active ingredients to achieve optimal dissolution efficiency in a short period of time and can reduce the damage to heat-sensitive substances caused by long-term high-temperature heating. At the same time, after acid treatment, it is easy to carry out selective separation in subsequent centrifugation, filtration and alkali precipitation steps. By changing the pH, non-target small molecules or impurities can be kept dissolved in the solution, and the target molecules can be precipitated or enriched in the desired form during the subsequent alkali adjustment, ensuring the coordination and unity of component conversion and purification between each step in the entire process.

[0029] Preferably, in step S300, the undissolved cell debris, cell wall residues and other insoluble impurities in the second mixture are separated by centrifugation and filtration to obtain a clarified first filtrate, thereby effectively removing large solid particles, preventing blockage or precipitation interference in subsequent process steps, and further improving the purity and stability of the extract; and the centrifugal treatment speed is controlled at 3000-5000 r / min and the time is 10-20 min, which can provide sufficient centrifugal force to sediment most of the insoluble particles to the bottom, and avoid excessive speed causing structural damage to heat-sensitive or shear-sensitive active molecules. Relatively moderate centrifugal speed and duration can promote the rapid stratification of small particles and macromolecular complexes without causing local overheating of the sample or precipitation of active components due to excessive shearing due to long-term ultra-high-speed operation.

[0030] Preferably, in step S400, the target active ingredient is further separated and enriched from the impurities by adding a secondary alkaline substance, followed by dialysis to remove small molecular impurities and salts, and finally a high-purity and stable Agaricus bisporus extract is prepared by reduced pressure concentration and freeze-drying; in S410, an alkaline substance is added dropwise to the first filtrate and the pH is adjusted to 6-8, which can promote the formation of insoluble precipitates in the environment of some active polysaccharides and polyphenols that are stable under acidic conditions after the addition of the alkaline substance. Stirring for 30-60 minutes and standing for 1-2 hours allows non-target impurities to continue to dissolve and remain in the supernatant, while the active ingredients gather at the bottom in the form of precipitates, thereby achieving preliminary enrichment and purification, removing some medium and low molecular weight impurities, and helping to change the structure of active molecules, further improving the biological activity and stability of the finished product in subsequent steps.

[0031] In the dialysis treatment at S420, the sediment is placed in a dialysis bag with a molecular weight cutoff of 1000 to 5000 Da, and the dialysis water is continuously replaced with deionized water 3 to 5 times. This can remove salts, acid hydrolysis products, and other small molecular impurities smaller than the cutoff range of the dialysis bag, retain active ingredients with molecular weights in the target range, and significantly improve the purity of the extract. At the same time, dialysis can also slowly restore the pH of the system to neutral, which is beneficial for subsequent stability and biological activity maintenance. In S430, vacuum concentration is carried out at -0.06 to -0.08 MPa and 40-60°C, which can remove most of the water at a lower temperature without destroying the heat-sensitive active ingredients. After the volume of the concentrate is reduced to 1 / (5 to 10) of the original volume, the extract is prepared into a dry powder for long-term storage. Step S400 achieves efficient separation, purification, and preparation of the Agaricus bisporus extract through the combined steps of alkaline precipitation, dialysis, vacuum concentration, and freeze-drying, ensuring the high purity, high activity, and good preservation of the product.

[0032] Example 1 This embodiment provides an Agaricus bisporus extract and a preparation method and application thereof, comprising the following steps: S100, weighing 500 g of fresh Agaricus bisporus slices, adding 3000 mL of deionized water, placing the mixture in a microwave reaction vessel, heating the mixture in a first step at 90° C. for 2 h, and in a second step at 120° C. for 3 h, adding sodium hydroxide to adjust the pH to 10, to obtain a first mixture; S200, after the first mixture is cooled, transfer it to an ultrasonic device, ultrasonicate it at 600W for 2 hours, add hydrochloric acid dropwise to adjust the pH value to 4.0, with a pulse time of 3:1, to obtain a second mixture; S300, centrifuging the second mixture at 4000 r / min for 15 minutes, collecting the supernatant and filtering it to obtain a first filtrate; S410, potassium hydroxide is added dropwise to the first filtrate to adjust the pH value to 7, stirring for 45 minutes, and standing for 1.5 hours to obtain a precipitate; S420, dialyzing the sediment in a dialysis bag with a molecular weight cutoff of 3000 Da for 18 hours, and changing the water four times during the dialysis treatment to obtain a dialyzed solution; S430, concentrating the dialyzed solution at 50° C. and −0.07 MPa to reduce the volume of the dialyzed solution to 1 / 8 of the original volume, and freeze-drying the solution to obtain an Agaricus bisporus extract.

[0033] Example 2 This embodiment provides an Agaricus bisporus extract and a preparation method and application thereof, comprising the following steps: S100, weighing 800 g of fresh Agaricus bisporus slices, adding 5000 mL of deionized water, placing the mixture in a microwave reaction vessel, heating the mixture in a first step at 90° C. for 2 h, and in a second step at 130° C. for 3 h, adding potassium hydroxide to adjust the pH to 12, to obtain a first mixture; S200, after the first mixture is cooled, transfer it to an ultrasonic device, ultrasonicate it at 800W for 2h, add acetic acid dropwise to adjust the pH value to 4.0, with a pulse time of 2:1, to obtain a second mixture; S300, centrifuging the second mixture at 4000 r / min for 15 minutes, collecting the supernatant and filtering it to obtain a first filtrate; S410, potassium hydroxide is added dropwise to the first filtrate to adjust the pH to 6, stirring for 45 minutes, and standing for 1.5 hours to obtain a precipitate; S420, dialyzing the sediment in a dialysis bag with a molecular weight cutoff of 3000 Da for 18 hours, and changing the water four times during the dialysis treatment to obtain a dialyzed solution; S430, concentrating the dialyzed solution at 50° C. and −0.07 MPa to reduce the volume of the dialyzed solution to 1 / 8 of the original volume, and freeze-drying the solution to obtain an Agaricus bisporus extract.

[0034] Example 3 This embodiment provides an Agaricus bisporus extract and a preparation method and application thereof, comprising the following steps: S100, weighing 800 g of fresh Agaricus bisporus slices, adding 4000 mL of deionized water, placing the mixture in a microwave reaction vessel, heating the mixture in a first step at 120° C. for 2 h, and in a second step at 180° C. for 2 h, adding sodium hydroxide to adjust the pH to 14, to obtain a first mixture; S200, after the first mixture is cooled, transfer it to an ultrasonic device, ultrasonicate it at 1000W for 2 hours, add hydrochloric acid dropwise to adjust the pH value to 2.5, with a pulse time of 3:1, to obtain a second mixture; S300, centrifuging the second mixture at 3000 r / min for 10 minutes, collecting the supernatant and filtering it to obtain a first filtrate; S410, adding potassium hydroxide dropwise to the first filtrate, adjusting the pH to 8, stirring for 60 minutes, and allowing to stand for 2 hours to obtain a precipitate; S420, dialyzing the sediment in a dialysis bag with a molecular weight cutoff of 1000 Da for 12 hours, and changing the water 5 times during the dialysis treatment to obtain a dialyzed solution; S430, concentrating the dialyzed solution at 40° C. and −0.06 MPa to reduce the volume of the dialyzed solution to 1 / 5 of the original volume, and freeze-drying the solution to obtain an Agaricus bisporus extract.

[0035] Example 4 This embodiment provides an Agaricus bisporus extract and a preparation method and application thereof, comprising the following steps: S100, weighing 500 g of fresh Agaricus bisporus slices, adding 5000 mL of deionized water, placing the mixture in a microwave reaction vessel, heating the mixture in a microwave oven at a first step temperature of 90° C. for 4 h, and in a second step temperature of 120° C. for 4 h, adding potassium hydroxide to adjust the pH to 8, to obtain a first mixture; S200, after the first mixture is cooled, transfer it to an ultrasonic device, ultrasonicate it at 300W for 8 hours, add acetic acid dropwise to adjust the pH value to 5.5, with a pulse time of 2:1, to obtain a second mixture; S300, centrifuging the second mixture at 3000 r / min for 20 minutes, collecting the supernatant and filtering it to obtain a first filtrate; S410, adding potassium hydroxide dropwise to the first filtrate to adjust the pH to 7, stirring for 30 minutes, and allowing to stand for 1 hour to obtain a precipitate; S420, dialyzing the sediment in a dialysis bag with a molecular weight cutoff of 5000 Da for 24 hours, and changing the water three times during the dialysis treatment to obtain a dialyzed solution; S430, concentrating the dialyzed solution at 60° C. and −0.08 MPa to reduce the volume of the dialyzed solution to 1 / 10 of the original volume, and freeze-drying the solution to obtain an Agaricus bisporus extract.

[0036] Comparative Example 1 This comparative example provides an Agaricus bisporus extract and a preparation method thereof. The specific steps are as shown in Example 1, except that, in step S200, no acidic substance is added for adjustment, that is, only ultrasonic treatment is performed.

[0037] Comparative Example 2 This comparative example provides a gel composite for a removable retainer. The specific steps are the same as those in Example 1, except that ultrasonic treatment is not performed in step S200, that is, only an acidic substance is added for adjustment.

[0038] Comparative Example 3 This comparative example provides a gel composite for a removable retainer, and the gel composite is purchased from outside.

[0039] Test data The extract of Example 1 was tested at a concentration of 125 μg / mL for the fluorescence intensity of zebrafish yolk sac fat and the staining intensity of intestinal and tail vascular fat. The extract of Example 2 was tested at a concentration of 62.5 μg / mL for the fluorescence intensity of zebrafish yolk sac fat and the staining intensity of intestinal and tail vascular fat. The results are as follows: Figure 1 、 Figure 2 As shown; it can be seen from the figure that the test results of Examples 1-2 prove that the present application has good fat-blocking and fat-reducing effects.

[0040] The extraction rate of Example 1 and Comparative Examples 1-2 was calculated. The same mass of Agaricus bisporus raw materials were subjected to the respective process treatments. Finally, the pure extract after concentration and freeze-drying was weighed. This value was divided by the original feed (calculated on a dry basis) to obtain the extraction rate. The results are shown in Table 1.

[0041] Table 1 The extraction rate of Example 1 was 25.36%, which was significantly higher than 5.17% of Comparative Example 1 and 15.42% of Comparative Example 2. This indicates that the preparation method adopted in Example 1 can obtain more dried extract from the same amount of Agaricus bisporus raw material than Comparative Examples 1 and 2, fully verifying the advantages of this method.

[0042] Throughout this specification, terms such as "one embodiment," "some embodiments," and "specific embodiments" mean that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0043] Although the present invention is disclosed as above, the present invention is not limited thereto. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the scope defined by the claims.

Claims

1. A method for preparing an Agaricus bisporus extract, characterized in that: The following steps are involved: S100, adding Agaricus bisporus slices to deionized water, heating with a microwave, and adding a first alkaline substance to adjust the pH value to obtain a first mixture; S200, adding an acidic substance while subjecting the first mixture to ultrasonic treatment to obtain a second mixture; S300, centrifuging and filtering the second mixture to obtain a first filtrate; S400, adding a second alkaline substance to the first filtrate, and performing stirring treatment, dialyzing treatment, and finally concentrating and freeze-drying treatment to obtain the Agaricus bisporus extract.

2. The preparation method according to claim 1, characterized in that Step S200 specifically includes: After the first mixture is cooled, ultrasonic treatment is performed at 300-1000 W for 2-8 hours, and an acidic substance is added dropwise to control the pH value of the first mixture to be 2.5-5.5, thereby obtaining a second mixture.

3. The preparation method according to claim 2, characterized in that The acidic substance includes at least one of hydrochloric acid and acetic acid.

4. The preparation method according to claim 1, characterized in that In step S100: The mass ratio of the Agaricus bisporus slices to the deionized water is 1:(5-10); The first alkaline substance includes at least one of sodium hydroxide and potassium hydroxide; The pH value of the first mixture is between 8 and 14; The microwave heating time is 4 to 8 hours, and the temperature is 90 to 180° C.

5. The preparation method according to claim 1, characterized in that The temperature of the microwave heating is controlled by gradient, including a first step temperature and a second step temperature; The temperature of the first step temperature ranges from 90 to 120°C, and the temperature of the second step temperature ranges from 120 to 180°C.

6. The preparation method according to claim 1, characterized in that In step S300, The rotation speed of the centrifugal treatment is 3000-5000 r / min, and the time is 10-20 min.

7. The preparation method according to claim 1, characterized in that Step S400 specifically includes: S410, dropwise adding the second alkaline substance solution to the first filtrate, adjusting the pH value to 6-8, performing the stirring treatment for 30-60 minutes, and then standing for 1-2 hours to obtain a precipitate; S420, placing the sediment in a dialysis bag with a molecular weight cut-off of 1000 to 5000 Da, performing the dialysis treatment for 12 to 24 hours, and replacing the dialysis water 3 to 5 times during the dialysis treatment to obtain a dialyzed solution; S430, performing the concentration treatment on the dialyzed solution under reduced pressure to reduce the volume of the dialyzed solution to 1 / (5-10) of the original volume, and performing the freeze-drying treatment to obtain the Agaricus bisporus extract.

8. The preparation method according to claim 7, characterized in that In step S410, the second alkaline substance includes at least one of sodium hydroxide and potassium hydroxide; In step S430, the vacuum degree in the decompression conditions is -0.06 to -0.08 MPa, and the temperature is 40 to 60°C.

9. An Agaricus bisporus extract, characterized in that The Agaricus bisporus extract is prepared by the preparation method according to any one of claims 1 to 8.

10. An application of Agaricus bisporus extract, characterized in that: Use of the Agaricus bisporus extract prepared by the preparation method according to any one of claims 1 to 8 in the preparation of foods or health products that inhibit fat absorption.

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