Edible mushroom extract and application of edible mushroom extract as antioxidant preparation in storage of coarse cereals

Abstract

The invention discloses an edible mushroom extract and application thereof as an antioxidant preparation in storage of coarse cereals. The invention provides application of an edible mushroom extract, namely application of the edible mushroom extract as an antioxidant. The invention discloses application of an edible mushroom extract in preparation of an antioxidant. The edible mushroom extract is applied to preparation of an additive for storing coarse cereals; the edible mushroom extract is used as an additive in storage of coarse cereals; the edible mushroom extract is applied to storage of coarse cereals as an additive with an anti-oxidation function. The edible mushroom extract is an agrocybe cylindracea polysaccharide extract or a pholiota adipose polyphenol extract or a grifola frondosa polyphenol extract. The edible fungus extract provided by the invention has good antioxidant activity, can inhibit oxidase systems of the coarse cereals so as to inhibit free fatty acid accumulation and / or malonaldehyde accumulation in the coarse cereals, and is free of any harmful component. The method is helpful for solving the practical problem that the coarse cereals are easy to oxidize in the storage and processing processes.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology and relates to an edible fungus extract and an application of the extract as an antioxidant preparation in the storage of grains. Background Art

[0002] Coarse grains generally refer to grains and legumes other than the five major crops of rice, wheat, corn, soybeans, and potatoes. These grains primarily include sorghum, millet, buckwheat, oats, quinoa, barley, broomcorn millet, sorghum foie gras, Job's tears, amaranth, kidney beans, mung beans, adzuki beans (red beans, red beans), broad beans, peas, cowpeas, lentils, and black beans. While their yields are lower than those of grain crops like rice, wheat, and corn, they offer a wide variety and rich nutrition, enriching our food sources.

[0003] During long-term storage of grains, key enzymes such as lipase (LPS), lipoxygenase (LOX), polyphenol oxidase (PPO), peroxidase (POD), and acyl-CoA oxidase (A-COX) work synergistically to catalyze a lipid oxidation chain reaction. This process leads to a significant increase in free fatty acid (FFA) content (ΔFFA ≥ 28%), accompanied by the accumulation of secondary oxidation products such as malondialdehyde (MDA) (MDA production increase > 20%), ultimately causing grain rancidity and deterioration, manifested by the development of a rancid odor, loss of nutrients, and a shortened shelf life.

[0004] Currently, synthetic antioxidants are increasingly being questioned due to their potential hazards and toxicity, while natural antioxidants are becoming a hot topic due to their environmental friendliness, safety, and nutritional benefits. Natural antioxidants, derived from plants, animals, and microorganisms, not only possess significant antioxidant properties but also possess a variety of biological activities, including anti-inflammatory, anti-aging, and immunomodulatory properties. Summary of the Invention

[0005] The present invention aims to provide an edible fungus extract and its application as an antioxidant preparation in the storage of grains.

[0006] The present invention provides applications of edible fungus extracts; the applications are as follows (I) or (II) or (III) or (IV) or (V):

[0007] (I) Application of edible fungus extracts as antioxidants;

[0008] (II) Application of edible fungus extracts in the preparation of antioxidants;

[0009] (III) Application of edible fungus extracts in the preparation of additives for grain storage;

[0010] (IV) Application of edible fungus extracts as additives in grain storage;

[0011] (V) Application of edible fungus extracts as additives with antioxidant function in grain storage;

[0012] The edible fungus extract is a polysaccharide extract of Agrocybe tumefaciens, a polyphenol extract of Pholiota adiposa, or a polyphenol extract of Grifola frondosa.

[0013] The present invention also provides an antioxidant, which comprises an edible fungus extract; the edible fungus extract is a tea tree agaric polysaccharide extract, a pholiotaep polyphenol extract, or a glycine max polyphenol extract.

[0014] The antioxidant has the following functions: scavenging hydroxyl radicals and / or scavenging superoxide anion radicals and / or scavenging ABTS radicals.

[0015] The present invention also provides an additive for storing grains, which comprises an edible fungus extract; the edible fungus extract is a tea tree agaric polysaccharide extract, a pholiota polyphenol extract, or a maitake mushroom polyphenol extract.

[0016] The additive plays the role of an antioxidant during the storage of grains. The antioxidant has the following functions: scavenging hydroxyl radicals and / or scavenging superoxide anion radicals and / or scavenging ABTS radicals.

[0017] During the storage of coarse grains, the additive performs the following functions: inhibiting the accumulation of free fatty acids and / or malondialdehyde in the coarse grains. During the storage of coarse grains, the additive performs the following functions: inhibiting the coarse grain oxidase system; the coarse grain oxidase system is lipase and / or lipoxygenase and / or polyphenol oxidase and / or peroxidase and / or peroxidase acyl-CoA oxidase. During the storage of coarse grains, the additive performs the following functions: reducing the free fatty acid content and / or malondialdehyde content in the coarse grains. During the storage of coarse grains, the additive performs the following functions: inhibiting the increase in lipase activity and / or inhibiting the increase in lipoxygenase activity and / or inhibiting the increase in peroxidase activity and / or inhibiting the increase in peroxidase acyl-CoA oxidase content.

[0018] When the edible fungus extract is used as an additive, 0.5-2g of the additive is added to every 100g of miscellaneous grains. When the edible fungus extract is used as an additive, 0.5-1.5g of the additive is added to every 100g of miscellaneous grains. When the edible fungus extract is used as an additive, 0.5g of the additive is added to every 100g of miscellaneous grains. When the edible fungus extract is used as an additive, 1g of the additive is added to every 100g of miscellaneous grains. When the edible fungus extract is used as an additive, 1.5g of the additive is added to every 100g of miscellaneous grains. When the edible fungus extract is used as an additive, 2g of the additive is added to every 100g of miscellaneous grains.

[0019] The present invention also protects an edible fungus extract, which is characterized in that the edible fungus extract is a tea tree agaric polysaccharide extract, a pholiotaep polyphenol extract, or a maitake mushroom polyphenol extract.

[0020] Any of the above-mentioned Agrocybe tumefaciens polysaccharide extracts is prepared using Agrocybe tumefaciens as the raw material using a water extraction-ethanol precipitation method. As an example, the water extraction temperature is 80-100°C. As an example, the water extraction temperature is 90°C. As an example, the water extraction time is 5-7 hours. As an example, the water extraction time is 6 hours.

[0021] Any of the above-mentioned Phellodendron floribunda polyphenol extracts is obtained by extraction with an ethanol-water solution using Phellodendron floribunda as a raw material. As an example, the ethanol-water solution is an ethanol-water solution with an ethanol volume percentage of 70%-90%. As an example, the ethanol-water solution is an ethanol-water solution with an ethanol volume percentage of 80%. As an example, the extraction is performed under microwave conditions. As an example, the extraction time is 4-6 minutes. As an example, the extraction time is 5 minutes. As an example, the power of the microwave is 600-800W. As an example, the power of the microwave is 700W.

[0022] Any of the above-mentioned Grifola frondosa polyphenol extracts is obtained by extracting Grifola frondosa using an ethanol-water solution. As an example, the ethanol-water solution is an ethanol-water solution having an ethanol volume percentage of 70%-90%. As an example, the ethanol-water solution is an ethanol-water solution having an ethanol volume percentage of 80%. As an example, the extraction is performed under microwave conditions. As an example, the extraction time is 4-6 minutes. As an example, the extraction time is 5 minutes. As an example, the microwave power is 600-800W. As an example, the microwave power is 700W.

[0023] The preparation method of any of the above-mentioned Agrocybe aegerita polysaccharide extracts comprises the following steps:

[0024] (1) Mixing Agrocybe oleifera with water, performing extraction, filtering, and collecting the filtrate;

[0025] (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 60-70° C. to obtain a concentrated solution;

[0026] (3) Add 3-5 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let it stand at 20-25°C for 10-14 hours. Then, centrifuge and discard the supernatant. Collect the precipitate and dry it at 50-60°C to constant weight to obtain a polysaccharide extract of Agrocybe oleifera.

[0027] The preparation method of any of the above-mentioned Pholiota adiposa polyphenol extracts comprises the following steps:

[0028] (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation;

[0029] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0030] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract.

[0031] The preparation method of any of the above-mentioned Grifola frondosa polyphenol extracts comprises the following steps:

[0032] (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant;

[0033] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0034] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

[0035] The present invention also provides a method for preparing a polysaccharide extract of Agrocybe edulis, comprising the following steps:

[0036] (1) Mixing Agrocybe oleifera with water, performing extraction, filtering, and collecting the filtrate;

[0037] (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 60-70° C. to obtain a concentrated solution;

[0038] (3) Add 3-5 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let it stand at 20-25°C for 10-14 hours. Then, centrifuge and discard the supernatant. Collect the precipitate and dry it at 50-60°C to constant weight to obtain a polysaccharide extract of Agrocybe oleifera.

[0039] The present invention also provides a method for preparing the Pholiota adiposa polyphenol extract, comprising the following steps:

[0040] (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation;

[0041] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0042] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract.

[0043] The present invention also provides a method for preparing a Grifola frondosa polyphenol extract, comprising the following steps:

[0044] (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant;

[0045] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0046] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

[0047] Specifically, the preparation method of the Agrocybe tumefaciens polysaccharide extract comprises the following steps:

[0048] (1) Mixing Agrocybe oleifera with water, extracting at 80-100°C for 5-7 hours, filtering, and collecting the filtrate;

[0049] (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 60-70° C. to obtain a concentrated solution;

[0050] (3) Add 3-5 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let it stand at 20-25°C for 10-14 hours. Then, centrifuge and discard the supernatant. Collect the precipitate and dry it at 50-60°C to constant weight to obtain a polysaccharide extract of Agrocybe oleifera.

[0051] Specifically, the preparation method of the Agrocybe tumefaciens polysaccharide extract comprises the following steps:

[0052] (1) Mixing Agrocybe oleifera with water, extracting at 90°C for 6 h, filtering, and collecting the filtrate;

[0053] (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 65° C. to obtain a concentrated solution;

[0054] (3) Add 4 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let it stand at 20-25°C for 12 hours. Then, centrifuge and discard the supernatant. Collect the precipitate and dry it at 55°C to constant weight to obtain the Agrocybe aegerita polysaccharide extract.

[0055] As an example, the centrifugal parameters are: 7000-9000 r / min, 8-12 min.

[0056] As an example, the centrifugal parameters are: 8000 r / min, 10 min.

[0057] As an example, in step (1), the ratio of Agrocybe aegerita to water is 8-12 g: 250 mL.

[0058] As an example, in step (1), the ratio of Agrocybe aegerita to water is 10 g:250 mL.

[0059] As an example, the filtering is performed using gauze.

[0060] As an example, the filtration is performed using 4 layers of gauze.

[0061] Specifically, the preparation method of the Agrocybe tumefaciens polysaccharide extract comprises the following steps:

[0062] (1) Take 10 g of Agrocybe aegerita, add 250 mL of deionized water, mix well, and extract in a 90°C water bath for 6 h. Filter with gauze and collect the filtrate.

[0063] (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 65° C. to obtain a concentrated solution;

[0064] (3) Add 4 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), stir evenly, and then let it stand at room temperature for 12 hours, then centrifuge at 8000 r / min for 10 minutes, discard the supernatant, collect the precipitate and dry it at 55°C to constant weight, and grind it into powder, which is the tea tree agaric polysaccharide extract.

[0065] The tea tree mushroom (the raw material tea tree mushroom) is tea tree mushroom powder. The tea tree mushroom (the raw material tea tree mushroom) is tea tree mushroom fruiting body powder. The tea tree mushroom powder is obtained by drying and crushing the tea tree mushroom fruiting bodies. The preparation method of the tea tree mushroom powder is as follows: take the tea tree mushroom fruiting bodies, dry them at 40°C to constant weight, then crush them and pass them through a 40-mesh sieve to obtain the tea tree mushroom powder.

[0066] Specifically, the preparation method of the Pholiota adiposa polyphenol extract comprises the following steps:

[0067] (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation;

[0068] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0069] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract.

[0070] Specifically, the preparation method of the Pholiota adiposa polyphenol extract comprises the following steps:

[0071] (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation;

[0072] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 55° C. to obtain a concentrated solution;

[0073] (3) The concentrated solution obtained in step (2) was dried at 55° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract.

[0074] In step (1), the centrifugal parameters may be: 5000-7000 r / min for 20-40 min.

[0075] In step (1), the centrifugal parameters may be: 6000 r / min for 30 min.

[0076] As an example, in step (1), the ratio of Phellodendron chinense and ethanol aqueous solution is: 4-6g:100mL.

[0077] As an example, in step (1), the ratio of Phellodendron chinense and ethanol aqueous solution is: 5g:100mL.

[0078] As an example, the ethanol aqueous solution is an ethanol aqueous solution with an ethanol volume percentage of 70%-90%.

[0079] As an example, the ethanol aqueous solution is an ethanol aqueous solution with an ethanol volume percentage of 80%.

[0080] As an example, the extraction is performed under microwave conditions.

[0081] As an example, the extraction time is 4-6 minutes.

[0082] As an example, the extraction time is 5 minutes.

[0083] As an example, the power of the microwave is 600-800W.

[0084] As an example, the power of the microwave is 700W.

[0085] Specifically, the preparation method of the Pholiota adiposa polyphenol extract comprises the following steps:

[0086] (1) 5 g of Phellodendron chinense was added to 100 mL of 80% (volume percentage) ethanol aqueous solution, mixed, and then extracted under microwave conditions (power of 700 W, time for 5 min), followed by centrifugation at 6000 rpm for 30 min, and the supernatant was collected;

[0087] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 55° C. to obtain a concentrated solution;

[0088] (3) The concentrated solution obtained in step (2) is dried at 55° C. to a constant weight and ground into powder to obtain the Pholiota adiposa polyphenol extract.

[0089] The phellodendron agarbatti (the raw material phellodendron agarbatti) is phellodendron agarbatti powder. The phellodendron agarbatti (the raw material phellodendron agarbatti) is phellodendron agarbatti fruiting body powder. The phellodendron agarbatti powder is obtained by drying and crushing the phellodendron agarbatti fruiting bodies. The preparation method of the phellodendron agarbatti powder is as follows: taking the phellodendron agarbatti fruiting bodies, drying them at 40°C to constant weight, then crushing and passing through a 40-mesh sieve to obtain the phellodendron agarbatti powder.

[0090] Specifically, the preparation method of the Grifola frondosa polyphenol extract comprises the following steps:

[0091] (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant;

[0092] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution;

[0093] (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

[0094] Specifically, the preparation method of the Grifola frondosa polyphenol extract comprises the following steps:

[0095] (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant;

[0096] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 55° C. to obtain a concentrated solution;

[0097] (3) The concentrated solution obtained in step (2) was dried at 55° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

[0098] In step (1), the centrifugal parameters may be: 5000-7000 r / min for 20-40 min.

[0099] In step (1), the centrifugal parameters may be: 6000 r / min for 30 min.

[0100] As an example, in step (1), the ratio of maitake mushroom and ethanol aqueous solution is: 4-6 g: 100 mL.

[0101] As an example, in step (1), the ratio of maitake mushroom and ethanol aqueous solution is: 5g:100mL.

[0102] As an example, the ethanol aqueous solution is an ethanol aqueous solution with an ethanol volume percentage of 70%-90%.

[0103] As an example, the ethanol aqueous solution is an ethanol aqueous solution with an ethanol volume percentage of 80%.

[0104] As an example, the extraction is performed under microwave conditions.

[0105] As an example, the extraction time is 4-6 minutes.

[0106] As an example, the extraction time is 5 minutes.

[0107] As an example, the power of the microwave is 600-800W.

[0108] As an example, the power of the microwave is 700W.

[0109] Specifically, the preparation method of the Grifola frondosa polyphenol extract comprises the following steps:

[0110] (1) 5 g of maitake mushrooms were added to 100 mL of 80% (volume percentage) ethanol aqueous solution, mixed, and then extracted under microwave conditions (power of 700 W, time for 5 min). The mixture was then centrifuged at 6000 rpm for 30 min, and the supernatant was collected.

[0111] (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 55° C. to obtain a concentrated solution;

[0112] (3) The concentrated solution obtained in step (2) is dried at 55° C. to a constant weight and ground into powder to obtain the Grifola frondosa polyphenol extract.

[0113] The maitake mushroom (the raw material) is maitake mushroom powder. The maitake mushroom (the raw material) is maitake mushroom fruiting body powder. The maitake mushroom powder is obtained by drying and pulverizing the maitake mushroom fruiting bodies. The preparation method of the maitake mushroom powder is as follows: taking the maitake mushroom fruiting bodies, drying them at 40°C to constant weight, then pulverizing and passing through a 40-mesh sieve to obtain the maitake mushroom powder.

[0114] As an example, any of the above-mentioned grains is millet and / or oats and / or quinoa and / or buckwheat.

[0115] As an example, any of the above-mentioned maitake mushrooms is Grifola frondosa, ACCC 52690.

[0116] As an example, any of the above mentioned Pholiota adiposa is Pholiota adiposa, ACCC 52033.

[0117] As an example, any of the above-mentioned tea tree mushrooms is tea tree mushroom (Agrocybe cylindracea), ACCC50913.

[0118] The present invention provides an edible fungus extract with excellent antioxidant activity, useful as an antioxidant. The edible fungus extract provided by the present invention can be used as a cereal additive to inhibit the oxidative enzyme system of the cereals, thereby inhibiting the accumulation of free fatty acids and / or malondialdehyde in the cereals. The extract is free of any harmful components. The present invention helps address the practical problem of susceptibility to oxidation of cereals during storage and processing. BRIEF DESCRIPTION OF THE DRAWINGS

[0119] Figure 1 It is a bar graph of the hydroxyl radical scavenging rate in Example 6.

[0120] Figure 2 It is a bar graph of the superoxide anion radical scavenging rate in Example 6.

[0121] Figure 3 This is a bar chart of the ABTS free radical scavenging rate in Example 6.

[0122] Figure 4 This is a bar graph showing the inhibition rate of the test substances on millet lipase in Example 8.

[0123] Figure 5 This is a bar graph showing the inhibition rates of the test substances on oat lipase in Example 8.

[0124] Figure 6 This is a bar graph showing the inhibition rate of the test substances on quinoa lipase in Example 8.

[0125] Figure 7 This is a bar graph showing the inhibition rates of the test substances on buckwheat lipase in Example 8.

[0126] Figure 8 This is a bar graph showing the inhibition rate of the test substances on millet lipoxygenase in Example 8.

[0127] Figure 9 This is a bar graph showing the inhibition rates of the test substances on oat lipoxygenase in Example 8.

[0128] Figure 10 This is a bar graph showing the inhibition rate of the test substances on quinoa lipoxygenase in Example 8.

[0129] Figure 11 This is a bar graph showing the inhibition rates of the test substances on buckwheat lipoxygenase in Example 8.

[0130] Figure 12 This is a bar graph showing the inhibition rate of the test substances on millet polyphenol oxidase in Example 8.

[0131] Figure 13 This is a bar graph showing the inhibition rates of the test substances on oat polyphenol oxidase in Example 8.

[0132] Figure 14 This is a bar graph showing the inhibition rate of the test substances on quinoa polyphenol oxidase in Example 8.

[0133] Figure 15 This is a bar graph showing the inhibition rates of the test substances on buckwheat polyphenol oxidase in Example 8.

[0134] Figure 16 This is a bar graph showing the inhibition rate of the test substances on millet peroxidase in Example 8.

[0135] Figure 17 This is a bar graph showing the inhibition rates of the test substances on oat peroxidase in Example 8.

[0136] Figure 18 This is a bar graph showing the inhibition rate of the test substances on quinoa peroxidase in Example 8.

[0137] Figure 19 This is a bar graph showing the inhibition rates of the test substances on buckwheat peroxidase in Example 8.

[0138] Figure 20 This is a bar graph showing the inhibition rate of the test substances on millet peroxidase acyl-CoA oxidase in Example 8.

[0139] Figure 21 This is a bar graph showing the inhibition rate of the test substances on oat peroxidase acyl-CoA oxidase in Example 8.

[0140] Figure 22 This is a bar graph showing the inhibition rate of the test substances on quinoa peroxidase acyl-CoA oxidase in Example 8.

[0141] Figure 23 This is a bar graph showing the inhibition rate of the test substances on buckwheat peroxidase acyl-CoA oxidase in Example 8. DETAILED DESCRIPTION

[0142] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0143] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in literature in the field or in accordance with product specifications. Materials and reagents used in the following examples are commercially available unless otherwise specified. Unless otherwise specified, the following examples were performed in triplicate, with three replicates for each experiment, and the results were averaged. Room temperature: 20-25°C. In the tables and figures, polysaccharides refer to polysaccharide extracts, proteins and polypeptides refer to protein and polypeptide extracts, and polyphenols refer to polyphenol extracts. In the tables, maitake mushroom powder refers to maitake mushroom fruiting body powder, phellodendron auricularia powder refers to phellodendron auricularia fruiting body powder, Agrocybe fasciata powder refers to Agrocybe fasciata fruiting body powder, maitake mushroom polyphenols refer to maitake mushroom polyphenol extracts, phellodendron auricularia polyphenols refer to phellodendron auricularia polyphenol extracts, and Agrocybe fasciata polysaccharides refer to Agrocybe fasciata polysaccharide extracts. In Examples 1, 2, and 3, sufficient quantities of product can be obtained through multiple preparations. NBT: nitroblue nitrotetrazolium chloride. ABTS free radical scavenging activity assay kit: Beijing Solebow Technology Co., Ltd., catalog number BC4775. Lipase (LPS) activity assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC2345. Lipoxygenase (LOX) activity assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC0325. Peroxidase (POD) activity assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC0095. Polyphenol oxidase (PPO) activity assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC0195. Peroxidase acyl-CoA oxidase (A-COX / AOX) ELISA kit: Beijing Qisong Biological Co., Ltd., catalog number QS47199. Free fatty acid (FFA) content assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC0595. Malondialdehyde (MDA) content assay kit: Beijing Solaibao Technology Co., Ltd., catalog number BC0025. The Agricultural Culture Collection of China (ACCC) has a website address of http: / / www.accc.org.cn. The public can directly order strains online. Corresponding strains can be obtained from the ACCC based on their serial numbers starting from the date of collection. The maitake mushroom used in the Examples is Grifola frondosa, which was collected by the ACCC on October 11, 2012, with the accession number ACCC 52690. The Pholiota adiposa used in the Examples is Pholiota adiposa, which was collected by the ACCC on January 3, 2008, with the accession number ACCC 52033.The oyster mushroom used in the examples is Hypsizigus marmoreus, which has been collected by the China Agricultural Microorganism Culture Collection Center with a deposit number of ACCC 51948. The nameko mushroom used in the examples is Pholiotanameko, also known as Pholiota nameko, which has been collected by the China Agricultural Microorganism Culture Collection Center on December 1, 1985 with a deposit number of ACCC 50159. The elm yellow mushroom used in the examples is Pleurotus citrinopileatus, which has been collected by the China Agricultural Microorganism Culture Collection Center on October 11, 2012 with a deposit number of ACCC 52666. The tea tree mushroom used in the examples is Agrocybe cylindracea, which has been collected by the China Agricultural Microorganism Culture Collection Center on April 10, 1999 with a deposit number of ACCC 50913. The fungus used in the examples is Auricularia polytricha, which was collected by the China Agricultural Microorganism Culture Collection Center on December 29, 2008, with a deposit number of ACCC 52258. The umbellata used in the examples is Grifola umbellata, which was collected by the China Agricultural Microorganism Culture Collection Center on August 1, 1995, with a deposit number of ACCC 50673.

[0144] Example 1. Preparation of polysaccharide extract (hot water extraction-ethanol precipitation method)

[0145] 1. Take 10g of the substance to be extracted, add 250mL of deionized water, mix well and extract in a 90℃ water bath for 6h. Filter with 4 layers of gauze while hot to remove impurities and collect the filtrate.

[0146] 2. The filtrate obtained in step 1 was concentrated by rotary evaporation at 65°C to obtain a concentrated solution.

[0147] 3. After completing step 2, add 4 times the volume of anhydrous ethanol to the concentrate, stir evenly, and then let it stand at room temperature for 12 hours, then centrifuge at 8000r / min for 10min, discard the supernatant, collect the precipitate and place it in a 55℃ oven to dry to constant weight, and then grind it in a mortar to obtain a powdered product, which is the polysaccharide extract.

[0148] Example 2. Preparation of Protein Peptide Extracts (Ammonium Sulfate Precipitation Method)

[0149] 1. Take 10 g of the substance to be extracted, add 250 mL of deionized water, mix well, then let it stand at 4 ° C for 12 hours, then centrifuge at 7000 r / min for 25 minutes, and collect the supernatant.

[0150] 2. Slowly add ammonium sulfate to the supernatant obtained in step 1 while stirring at 4°C until the saturation reaches 80% (i.e., add 516 g of ammonium sulfate per 1 L of supernatant). Let it stand at -4°C for 4 h, then centrifuge at 4°C and 7000 rpm for 25 min, and discard the supernatant.

[0151] 3. After completing step 2, dissolve the precipitate with a small amount of deionized water, and then transfer it to a dialysis bag with a molecular weight cutoff of 3000Da. Then place the dialysis bag in a container and dialyze with running tap water under the faucet for 4 hours to remove small molecular impurities. Then transfer the dialysis bag to deionized water and dialyze at 4°C for 12 hours. Then collect the liquid phase in the dialysis bag, centrifuge it at 4°C and 7000r / min for 10 minutes, and discard the precipitate.

[0152] 4. After completing step 3, the supernatant was placed in a -80°C refrigerator and frozen for 48 hours, and then freeze-dried and ground in a mortar to obtain a powdered product, which is the protein polypeptide extract.

[0153] Example 3. Preparation of polyphenol extract (microwave-assisted extraction)

[0154] 1. Take 5 g of the substance to be extracted, add 100 mL of 80% (volume percentage) ethanol aqueous solution, mix well, and then perform microwave extraction (microwave extraction parameters: power 700 W, time 5 min), then centrifuge at 6000 rpm for 30 min, and collect the supernatant.

[0155] 2. The supernatant obtained in step 1 was concentrated by rotary evaporation at 55°C to obtain a concentrated solution.

[0156] 3. The concentrated solution obtained in step 2 was placed in an oven at 55°C and dried to constant weight, and then ground in a mortar to obtain a powdery product, which is the polyphenol extract.

[0157] Example 4: Preparation of multiple active ingredients from multiple raw materials

[0158] Take the fruiting bodies of Grifola frondosa, dry them at 40°C to constant weight, then grind them and pass them through a 40-mesh sieve to obtain Grifola frondosa fruiting body powder. Using the Grifola frondosa fruiting body powder as the substance to be extracted, the method of Example 1 is used to prepare a polysaccharide extract to obtain a Grifola frondosa polysaccharide extract. Using the Grifola frondosa fruiting body powder as the substance to be extracted, the method of Example 2 is used to prepare a protein polypeptide extract to obtain a Grifola frondosa protein polypeptide extract. Using the Grifola frondosa fruiting body powder as the substance to be extracted, the method of Example 3 is used to prepare a polyphenol extract to obtain a Grifola frondosa polyphenol extract.

[0159] Take the fruiting bodies of Pholiota adzuki, dry them at 40°C to constant weight, then crush and pass through a 40-mesh sieve to obtain a Pholiota adzuki fruiting body powder. Using the Pholiota adzuki fruiting body powder as the substance to be extracted, a polysaccharide extract was prepared using the method of Example 1 to obtain a Pholiota adzuki polysaccharide extract. Using the fruiting body powder of Pholiota adzuki as the substance to be extracted, a protein polypeptide extract was prepared using the method of Example 2 to obtain a Pholiota adzuki protein polypeptide extract. Using the fruiting body powder of Pholiota adzuki as the substance to be extracted, a polyphenol extract was prepared using the method of Example 3 to obtain a Pholiota adzuki polyphenol extract.

[0160] Take the fruiting body of the shimeji mushroom, dry it at 40°C to constant weight, then crush it and pass it through a 40-mesh sieve to obtain the fruiting body powder of the shimeji mushroom. Using the fruiting body powder of the shimeji mushroom as the substance to be extracted, the polysaccharide extract is prepared by the method of Example 1 to obtain the shimeji mushroom polysaccharide extract. Using the fruiting body powder of the shimeji mushroom as the substance to be extracted, the protein polypeptide extract is prepared by the method of Example 2 to obtain the shimeji mushroom protein polypeptide extract. Using the fruiting body powder of the shimeji mushroom as the substance to be extracted, the polyphenol extract is prepared by the method of Example 3 to obtain the shimeji mushroom polyphenol extract.

[0161] Take the fruiting bodies of Pleurotus ostreatus, dry them at 40°C to constant weight, then pulverize and pass through a 40-mesh sieve to obtain Pleurotus ostreatus fruiting body powder. Using the Pleurotus ostreatus fruiting body powder as the material to be extracted, prepare a polysaccharide extract using the method of Example 1 to obtain a Pleurotus ostreatus polysaccharide extract. Using the Pleurotus ostreatus fruiting body powder as the material to be extracted, prepare a protein polypeptide extract using the method of Example 2 to obtain a Pleurotus ostreatus protein polypeptide extract. Using the Pleurotus ostreatus fruiting body powder as the material to be extracted, prepare a polyphenol extract using the method of Example 3 to obtain a Pleurotus ostreatus polyphenol extract.

[0162] Take the fruiting body of Pleurotus citriodora, dry it at 40°C to constant weight, then crush it and pass it through a 40-mesh sieve to obtain the fruiting body powder of Pleurotus citriodora. Using the fruiting body powder of Pleurotus citriodora as the substance to be extracted, the polysaccharide extract is prepared by the method of Example 1 to obtain the Pleurotus citriodora polysaccharide extract. Using the fruiting body powder of Pleurotus citriodora as the substance to be extracted, the protein polypeptide extract is prepared by the method of Example 2 to obtain the Pleurotus citriodora protein polypeptide extract. Using the fruiting body powder of Pleurotus citriodora as the substance to be extracted, the polyphenol extract is prepared by the method of Example 3 to obtain the Pleurotus citriodora polyphenol extract.

[0163] Take the fruiting body of Agrocybe tumefaciens, dry it at 40°C to constant weight, then crush it and pass it through a 40-mesh sieve to obtain Agrocybe tumefaciens fruiting body powder. Using the Agrocybe tumefaciens fruiting body powder as the substance to be extracted, a polysaccharide extract was prepared using the method of Example 1 to obtain Agrocybe tumefaciens polysaccharide extract. Using the fruiting body powder of Agrocybe tumefaciens as the substance to be extracted, a protein polypeptide extract was prepared using the method of Example 2 to obtain Agrocybe tumefaciens protein polypeptide extract. Using the fruiting body powder of Agrocybe tumefaciens as the substance to be extracted, a polyphenol extract was prepared using the method of Example 3 to obtain Agrocybe tumefaciens polyphenol extract.

[0164] Take the fruiting body of Auricularia auricularia, dry it at 40°C to constant weight, then crush it and pass it through a 40-mesh sieve to obtain the fruiting body powder of Auricularia auricularia. Using the fruiting body powder of Auricularia auricularia as the substance to be extracted, prepare a polysaccharide extract by the method of Example 1 to obtain the Auricularia auricularia polysaccharide extract. Using the fruiting body powder of Auricularia auricularia as the substance to be extracted, prepare a protein polypeptide extract by the method of Example 2 to obtain the Auricularia auricularia protein polypeptide extract. Using the fruiting body powder of Auricularia auricularia as the substance to be extracted, prepare a polyphenol extract by the method of Example 3 to obtain the Auricularia auricularia polyphenol extract.

[0165] Take the fruiting bodies of Polyporus umbellatus, dry them at 40°C to constant weight, then crush and pass through a 40-mesh sieve to obtain Polyporus umbellatus fruiting body powder. Using the Polyporus umbellatus fruiting body powder as the substance to be extracted, a polysaccharide extract was prepared using the method of Example 1 to obtain a Polyporus umbellatus polysaccharide extract. Using the fruiting body powder of Polyporus umbellatus as the substance to be extracted, a protein polypeptide extract was prepared using the method of Example 2 to obtain a Polyporus umbellatus protein polypeptide extract. Using the fruiting body powder of Polyporus umbellatus as the substance to be extracted, a polyphenol extract was prepared using the method of Example 3 to obtain a Polyporus umbellatus polyphenol extract.

[0166] Example 5. Antioxidant Effect Detection Method

[0167] Free radical scavenging ability is an important indicator for evaluating antioxidant activity, which can directly reflect the ability of the test substance to block the lipid oxidation chain reaction.

[0168] Test solution: Dissolve the test substance in deionized water to a concentration of 2 mg / mL.

[0169] 1. Determination of hydroxyl radical scavenging rate

[0170] To 0.5 mL of 6 mmol / L FeSO4 aqueous solution, add 0.5 mL of the test solution, then add 0.5 mL of 6 mmol / L H2O2 aqueous solution, shake well and let it stand for 10 minutes, then add 0.5 mL of salicylic acid-ethanol solution (salicylic acid-ethanol solution: dissolve salicylic acid in anhydrous ethanol to make the concentration of salicylic acid 6 mmol / L, that is, salicylic acid-ethanol solution), shake well and let it stand at room temperature for 30 minutes, then take a sample and detect the absorbance at a wavelength of 510 nm to obtain the absorbance A1.

[0171] Replace salicylic acid with an equal volume of deionized water and repeat the above steps to obtain absorbance A2.

[0172] Replace the test solution with deionized water and repeat the above steps to obtain the absorbance A0.

[0173] The hydroxyl radical scavenging rate is represented by R, and the unit is %.

[0174]

[0175] 2. Determination of superoxide anion free radical scavenging rate

[0176] Preheat 2.5 mL of Tris-HCl buffer (pH 8.2, 0.1 mol / L) in a 25°C water bath for 20 minutes. Then, add 0.2 mL of the test solution, 0.6 mL of a 0.98 mmol / L NBT aqueous solution, and 0.3 mL of a 10 mmol / L pyrogallol aqueous solution. Mix thoroughly and react in a 25°C water bath for 4 minutes. Immediately, add 0.1 mL of an 8 mol / L HCl aqueous solution to terminate the reaction. Samples are then collected and the absorbance at 530 nm is measured to obtain the absorbance A1.

[0177] Replace the test solution with deionized water and repeat the above steps to obtain the absorbance A0.

[0178] The superoxide anion radical scavenging rate is expressed as O and the unit is %.

[0179]

[0180] 3. ABTS free radical scavenging rate determination

[0181] Take the test solution (as the sample solution) and use the ABTS free radical scavenging ability detection kit for determination (follow the instructions). The result is the ABTS free radical scavenging rate, and the unit is %.

[0182] Example 6: Comparison of the antioxidant effects of 24 test substances

[0183] The test substances were: maitake mushroom polysaccharide extract, maitake mushroom protein polypeptide extract, maitake mushroom polyphenol extract, phellodendron polysaccharide extract, phellodendron protein polypeptide extract, phellodendron polyphenol extract, shimeji mushroom polysaccharide extract, shimeji mushroom protein polypeptide extract, shimeji mushroom polyphenol extract, nameko agaricus polysaccharide extract, nameko agaricus protein polypeptide extract, nameko agaricus polyphenol extract, pleurotus citriodora polysaccharide extract, pleurotus citriodora protein polypeptide extract, pleurotus citriodora polyphenol extract, tea tree agaricus polysaccharide extract, tea tree agaricus protein polypeptide extract, tea tree agaricus polyphenol extract, Auricularia auricularia polysaccharide extract, Auricularia auricularia protein polypeptide extract, Auricularia auricularia polyphenol extract, Polyporus umbellatus polysaccharide extract, Polyporus umbellatus protein polypeptide extract, or Polyporus umbellatus polyphenol extract. All the test substances were prepared in Example 4.

[0184] Take the test sample and detect the hydroxyl radical scavenging rate according to the method of step 1 of Example 5 (the results are shown in Tables 1 and Figure 1 ), the superoxide anion radical scavenging rate was detected according to the method of step 2 of Example 5 (the results are shown in Tables 2 and Figure 2 ), the ABTS free radical scavenging rate was detected according to the method of step 3 of Example 5 (the results are shown in Tables 3 and Figure 3 ). The hydroxyl radical scavenging rate of the tea tree agrocybe polysaccharide extract was the highest, which was 70.89±1.24%. The superoxide anion radical scavenging rate of the tea tree agrocybe polysaccharide extract was the highest, which was 31.52±0.14%. The superoxide anion radical scavenging rates of the maitake mushroom polyphenol extract and the phellodendron polyphenol extract were 27.49±0.36% and 27.74±0.13%, respectively. The ABTS radical scavenging rate of the tea tree agrocybe polysaccharide extract was the highest, which was 61.26±1.21%. The ABTS radical scavenging rate of the phellodendron polyphenol extract was 55.86±0.32%. According to the results in Tables 1 to 3, the tea tree agrocybe polysaccharide extract, the maitake mushroom polyphenol extract and the phellodendron polyphenol extract had the best effects.

[0185] Table 1 Hydroxyl radical scavenging rate (%)

[0186] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 65.01±0.25 57.82±0.39 61.20±0.86 49.43±0.44 50.93±1.39 70.89±1.24 33.27±1.83 56.76±1.21 Protein Peptides 46.42±0.50 60.14±0.64 61.88±0.11 53.66±0.47 39.56±0.98 28.99±0.52 29.49±0.04 41.02±1.09 polyphenols 62.62±0.79 63.00±1.15 58.99±0.48 58.07±0.97 59.93±0.48 61.63±0.48 59.90±0.38 49.55±1.13

[0187] Table 2 Superoxide anion radical scavenging rate (%)

[0188] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 24.86±0.11 25.09±0.71 19.87±0.32 23.10±0.82 11.49±0.99 31.52±0.14 21.63±0.51 21.14±0.28 Protein Peptides 24.82±0.27 21.06±0.91 23.94±0.29 23.12±0.30 18.73±0.69 26.71±0.53 26.55±0.31 11.49±0.49 polyphenols 27.49±0.36 27.74±0.13 21.23±0.37 15.64±0.39 21.53±0.04 18.13±0.67 11.59±1.44 15.23±0.23

[0189] Table 3 ABTS free radical scavenging rate (%)

[0190]

[0191]

[0192] Example 7: Method for Detecting the Inhibitory Efficacy on Cereal Oxidase

[0193] The test grains were ground into powder using a grinder and passed through a 60-mesh sieve to obtain grain powder.

[0194] Test sample: Mix grain powder and test substance (the ratio of grain powder to test substance is 0.1g:2mg), stir and mix thoroughly.

[0195] Control sample: multi-grain powder.

[0196] Lipase inhibition assay: Use a lipase (LPS) activity assay kit according to the instructions. Sample solution preparation: Take 0.1 g of sample (test sample or control sample), add 1.0 mL of Reagent 1 (provided in the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Then, centrifuge at 4°C, 12,000 rpm, for 15 minutes. The supernatant is the sample solution. The results are the LPS activity of the test sample (U / g) and the LPS activity of the control sample (U / g). Lipase inhibition rate of the test substance = (LPS activity of the control sample - LPS activity of the test sample) / LPS activity of the control sample × 100%.

[0197] Lipoxygenase Inhibition Assay: Use a lipoxygenase (LOX) activity assay kit according to the manufacturer's instructions. Sample preparation: Take 0.1 g of sample (test sample or control sample), add 1 mL of extract (provided with the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C, 12,000 rpm, for 20 minutes. Collect the supernatant, which is the sample solution. Results are the LOX activity (U / g) of the test sample and the LOX activity (U / g) of the control sample. Lipoxygenase inhibition rate of the test substance = (LOX activity of the control sample - LOX activity of the test sample) / LOX activity of the control sample × 100%.

[0198] To test the inhibitory potency against polyphenol oxidase: Use a polyphenol oxidase (PPO) activity assay kit according to the instructions. Sample solution preparation: Take 0.1 g of sample (test sample or control sample), add 1 mL of the extract (provided by the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Then, centrifuge at 4°C and 12,000 rpm for 10 minutes. The supernatant is the sample solution. Results are the PPO activity (U / g) of the test sample and the PPO activity (U / g) of the control sample. The inhibitory rate of the test substance against polyphenol oxidase = (PPO activity of the control sample - PPO activity of the test sample) / PPO activity of the control sample × 100%.

[0199] Peroxidase inhibition test: Use a peroxidase (POD) activity test kit and follow the instructions. Sample solution preparation method: Take 0.1g of sample (test sample or control sample), add 1mL of extract (provided by the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Then centrifuge at 4°C and 12,000 rpm for 10 minutes. Take the supernatant, which is the sample solution. The results are the POD activity (U / g) of the test sample and the POD activity (U / g) of the control sample. The inhibition rate of the test substance on peroxidase = (POD activity of the control sample - POD activity of the test sample) ÷ POD activity of the control sample × 100%.

[0200] To test the inhibitory potency against peroxidase acyl-CoA oxidase, use a peroxidase acyl-CoA oxidase (A-COX / AOX) ELISA kit according to the manufacturer's instructions. Sample preparation: Take 0.1 g of sample (test sample or control sample), add 1 mL of normal saline, mix thoroughly, and incubate in an ice-water bath for 2 hours. Then, centrifuge at 4°C, 3000 rpm, and collect the supernatant, which is the sample solution. Results are the A-COX content (ng / g) of the test sample and the A-COX content (ng / g) of the control sample. The inhibition rate of the test substance against peroxidase acyl-CoA oxidase = (A-COX content of the control sample - A-COX content of the test sample) / A-COX content of the control sample × 100%.

[0201] Example 8: Comparison of the inhibitory effects of 24 test substances on cereal oxidases

[0202] The test substances were: maitake mushroom polysaccharide extract, maitake mushroom protein polypeptide extract, maitake mushroom polyphenol extract, phellodendron polysaccharide extract, phellodendron protein polypeptide extract, phellodendron polyphenol extract, shimeji mushroom polysaccharide extract, shimeji mushroom protein polypeptide extract, shimeji mushroom polyphenol extract, nameko agaricus polysaccharide extract, nameko agaricus protein polypeptide extract, nameko agaricus polyphenol extract, pleurotus citriodora polysaccharide extract, pleurotus citriodora protein polypeptide extract, pleurotus citriodora polyphenol extract, tea tree agaricus polysaccharide extract, tea tree agaricus protein polypeptide extract, tea tree agaricus polyphenol extract, Auricularia auricularia polysaccharide extract, Auricularia auricularia protein polypeptide extract, Auricularia auricularia polyphenol extract, Polyporus umbellatus polysaccharide extract, Polyporus umbellatus protein polypeptide extract, or Polyporus umbellatus polyphenol extract. All the test substances were prepared in Example 4.

[0203] The tested grains were (all commercially available): millet, oats, quinoa or buckwheat.

[0204] The test substance was taken and its inhibitory effect on the oxidative enzyme system of grains was detected according to the method of Example 7. The inhibition rate of the test substance on millet lipase is shown in Table 4 and Figure 4 The inhibition rates of the test substances on oat lipase are shown in Tables 5 and Figure 5 The inhibition rates of the test substances on quinoa lipase are shown in Tables 6 and Figure 6 The inhibition rates of the test substances on buckwheat lipase are shown in Tables 7 and Figure 7 The inhibition rates of the test substances on millet lipoxygenase are shown in Table 8 and Figure 8 The inhibition rates of the test substances on oat lipoxygenase are shown in Table 9 and Figure 9 The inhibition rates of the test substances on quinoa lipoxygenase are shown in Tables 10 and Figure 10 The inhibition rates of the test substances on buckwheat lipoxygenase are shown in Tables 11 and Figure 11 The inhibition rates of the test substances on millet polyphenol oxidase are shown in Table 12 and Figure 12The inhibition rates of the test substances on oat polyphenol oxidase are shown in Tables 13 and Figure 13 The inhibition rates of the test substances on quinoa polyphenol oxidase are shown in Tables 14 and Figure 14 The inhibition rates of the test substances on buckwheat polyphenol oxidase are shown in Tables 15 and Figure 15 The inhibition rates of the test substances on millet peroxidase are shown in Table 16 and Figure 16 The inhibition rates of the test substances on oat peroxidase are shown in Tables 17 and Figure 17 The inhibition rates of the test substances on quinoa peroxidase are shown in Tables 18 and Figure 18 The inhibition rates of the test substances on buckwheat peroxidase are shown in Tables 19 and Figure 19 The inhibition rates of the test substances on millet peroxidase acyl-CoA oxidase are shown in Table 20 and Figure 20 The inhibition rates of the test substances on oat peroxidase acyl-CoA oxidase are shown in Table 21 and Figure 21 The inhibition rates of the test substances on quinoa peroxidase acyl-CoA oxidase are shown in Table 22 and Figure 22 The inhibition rates of the test substances on buckwheat peroxidase acyl-CoA oxidase are shown in Table 23 and Figure 23 According to the results in Tables 4 to 23, the polysaccharide extract of Agrocybe tumefaciens, the polyphenol extract of Grifola frondosa, and the polyphenol extract of Pholiota adiposa had the best effects.

[0205] Table 4 Inhibition rate of the test substances on millet lipase (%)

[0206] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 22.58±0.27 14.53±0.22 28.48±0.54 21.98±0.19 14.61±0.38 31.44±0.41 10.99±0.12 4.82±0.26 Protein Peptides 16.90±0.71 21.57±0.51 17.36±0.38 7.51±0.38 16.20±0.18 14.28±0.23 17.07±0.50 16.57±0.24 polyphenols 29.97±0.33 31.13±0.24 16.68±0.20 19.53±0.44 21.79±0.52 15.66±0.33 14.46±0.35 13.82±0.35

[0207] Table 5 Inhibition rate of test substances on oat lipase (%)

[0208] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 32.62±0.35 46.33±0.25 37.11±0.41 18.69±0.25 36.53±0.48 45.43±0.35 9.34±0.45 4.86±0.27 Protein Peptides 22.08±0.29 38.05±0.56 12.59±0.36 17.30±0.39 21.87±0.52 29.56±0.33 8.14±0.48 12.42±0.31 polyphenols 48.52±0.15 48.20±0.09 35.05±0.38 37.15±0.05 29.37±0.49 17.35±0.28 13.98±0.44 14.42±0.37

[0209] Table 6 Inhibition rate of the test substances on quinoa lipase (%)

[0210] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 16.99±0.38 26.14±0.26 10.46±0.55 8.37±0.24 30.62±0.34 32.56±0.22 12.09±0.02 11.63±0.45 Protein Peptides 19.89±0.13 18.32±0.42 26.79±0.22 9.36±0.35 24.85±0.35 11.68±0.22 17.25±0.40 24.49±0.37 polyphenols 36.48±0.30 36.48±0.09 12.83±0.52 27.27±0.07 23.79±0.13 27.37±0.08 30.25±0.56 8.52±0.26

[0211] Table 7 Inhibition rate of the test substances on buckwheat lipase (%)

[0212] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 15.14±0.23 16.52±0.41 6.87±0.28 7.43±0.18 10.27±0.32 29.65±0.32 22.83±0.31 16.31±0.13 Protein Peptides 15.34±0.06 17.71±0.19 19.15±0.31 13.02±0.21 8.19±0.03 15.90±0.19 12.20±0.37 11.27±0.18 polyphenols 26.48±0.24 30.64±0.23 7.00±0.12 11.31±0.12 17.11±0.41 18.45±0.38 13.90±0.35 11.25±0.44

[0213] Table 8 Inhibition rate of the test substances on millet lipoxygenase (%)

[0214] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 29.66±2.76 40.62±0.01 21.87±0.00 28.12±5.42 46.88±5.41 53.12±5.41 21.87±0.01 15.62±5.41 Protein Peptides 46.87±5.42 31.25±0.00 46.87±5.42 21.87±0.00 28.13±5.41 18.75±5.41 15.62±5.41 3.12±0.01 polyphenols 65.62±5.42 68.75±0.00 37.50±5.41 6.25±5.41 3.13±0.00 46.88±5.41 25.00±5.42 18.75±5.41

[0215] Table 9 Inhibition rate of test substances on oat lipoxygenase (%)

[0216] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 15.26±1.13 36.36±0.00 45.45±3.94 22.73±0.00 22.73±0.00 72.72±3.94 13.64±3.94 6.82±3.94 Protein Peptides 34.09±3.93 29.55±0.00 27.28±3.94 45.45±3.94 31.82±3.93 54.55±3.94 38.63±3.94 54.55±3.94 polyphenols 68.18±3.93 77.27±0.00 40.91±3.94 40.91±3.94 43.18±0.00 50.00±0.00 38.63±3.94 18.18±3.94

[0217] Table 10 Inhibition rate of the test substances on quinoa lipoxygenase (%)

[0218] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 33.37±1.31 17.39±3.77 45.65±0.00 41.30±3.76 36.96±3.76 52.17±0.00 17.39±3.77 10.87±3.76 Protein Peptides 15.22±3.77 34.78±3.76 45.65±0.00 41.30±3.76 52.17±0.00 15.22±3.77 21.74±3.76 8.69±3.76 polyphenols 56.52±3.77 58.70±0.00 41.30±3.76 45.65±0.00 13.04±0.00 52.17±0.00 13.04±0.00 45.65±0.00

[0219] Table 11 Inhibition rate of the test substances on buckwheat lipoxygenase (%)

[0220] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 26.81±2.13 27.91±4.03 16.28±4.02 6.98±4.03 20.93±4.03 44.18±4.03 16.28±4.02 9.30±4.03 Protein Peptides 34.88±4.02 13.95±4.02 16.28±4.02 13.95±4.02 9.30±4.03 34.88±4.02 9.30±4.03 18.60±0.00 polyphenols 44.18±4.03 51.16±4.03 25.58±0.00 39.53±0.00 18.60±0.00 37.21±4.02 20.93±4.03 20.93±4.03

[0221] Table 12 Inhibition rate of millet polyphenol oxidase by the test substances (%)

[0222] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 44.95±0.14 37.80±0.38 25.97±0.75 24.90±0.38 35.99±2.86 69.02±0.87 10.52±0.65 9.78±2.35 Protein Peptides 5.09±0.99 37.84±0.25 1.23±0.57 10.11±0.71 51.31±1.11 44.04±0.25 58.58±1.86 30.81±1.49 polyphenols 71.33±0.14 67.05±1.22 36.56±0.28 0.90±0.74 46.84±1.48 27.20±0.14 20.05±2.53 10.11±3.46

[0223] Table 13 Inhibition rate of test substances on oat polyphenol oxidase (%)

[0224] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 33.93±0.38 23.43±0.44 37.26±1.17 12.42±1.15 39.69±1.76 59.28±0.39 13.06±0.59 8.06±0.22 Protein Peptides 35.72±0.59 37.26±1.60 11.52±3.65 22.92±0.44 34.06±0.44 29.32±0.77 18.57±0.39 28.43±2.44 polyphenols 59.54±2.12 50.58±0.80 38.16±0.39 30.60±3.65 19.59±1.17 18.44±0.44 26.89±3.11 19.33±0.39

[0225] Table 14 Inhibition rate of the test substances on quinoa polyphenol oxidase (%)

[0226] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 15.76±0.81 23.10±0.70 13.90±1.22 18.23±1.55 37.15±2.07 53.77±2.41 5.15±0.41 19.92±2.79 Protein Peptides 38.12±1.46 12.84±1.01 8.87±0.66 34.50±1.16 45.99±1.46 37.95±2.87 10.90±1.37 7.19±1.47 polyphenols 53.15±2.22 59.52±2.54 44.84±2.76 40.69±3.36 23.98±2.50 25.40±2.67 14.26±2.13 12.14±2.01

[0227] Table 15 Inhibition rate of buckwheat polyphenol oxidase by the test substances (%)

[0228] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 11.78±1.27 6.89±2.70 37.32±0.49 19.24±3.57 20.21±3.92 52.57±1.64 24.71±2.02 32.56±3.91 Protein Peptides 26.96±2.13 21.43±2.88 39.13±3.74 47.30±0.58 8.69±1.50 42.99±0.62 34.43±2.22 21.17±1.18 polyphenols 48.65±1.39 58.04±1.06 22.52±2.06 13.51±2.23 4.83±0.51 13.64±1.06 23.04±2.84 5.21±1.86

[0229] Table 16 Inhibition rate of test substances on millet peroxidase (%)

[0230] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 16.19±0.58 16.19±1.27 11.97±0.88 21.75±1.05 10.62±0.77 33.22±1.34 5.23±0.58 3.37±0.51 Protein Peptides 4.38±0.51 7.93±0.51 25.80±0.58 10.12±0.58 14.00±0.51 14.67±0.77 9.27±2.11 7.93±1.01 polyphenols 33.39±1.05 40.64±1.05 12.82±1.05 14.50±0.51 16.86±0.58 16.53±0.51 20.74±0.29 6.75±1.27

[0231] Table 17 Inhibition rate of test substances on oat peroxidase (%)

[0232] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 15.23±1.43 21.72±0.35 14.76±0.41 9.21±0.54 17.00±1.24 24.68±0.20 16.65±0.20 6.14±0.94 Protein Peptides 17.59±0.54 13.58±0.35 9.80±0.89 11.57±0.41 18.42±0.89 22.08±0.35 11.45±0.61 8.62±0.61 polyphenols 26.21±1.02 30.46±1.08 9.09±0.54 10.04±0.71 4.84±0.41 11.33±1.64 16.06±0.35 5.31±1.08

[0233] Table 18 Inhibition rate of test substances on quinoa peroxidase (%)

[0234] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 15.23±0.55 11.38±0.55 19.27±0.32 6.42±0.55 13.39±0.32 26.61±0.32 12.11±0.32 14.50±0.84 Protein Peptides 11.38±0.55 13.94±0.84 12.84±0.64 19.27±0.32 21.47±0.32 12.48±0.55 15.60±0.84 8.81±0.32 polyphenols 23.49±0.95 24.40±1.27 15.60±1.15 11.93±1.10 13.76±0.32 20.18±0.95 11.93±0.55 13.94±0.64

[0235] Table 19 Inhibition rate of test substances on buckwheat peroxidase (%)

[0236] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 12.19±0.29 13.34±1.03 20.92±0.49 17.46±0.49 8.40±0.29 26.19±0.29 14.83±1.03 3.95±1.59 Protein Peptides 12.19±0.75 20.43±0.49 15.65±0.29 20.26±0.29 18.62±0.29 18.45±0.49 5.93±0.75 14.17±0.57 polyphenols 24.22±0.75 24.22±0.75 15.98±0.49 12.52±0.49 20.76±0.29 17.13±1.03 13.18±0.75 2.47±0.75

[0237] Table 20 Inhibition rate of the test substances on millet peroxidase acyl-CoA oxidase (%)

[0238]

[0239]

[0240] Table 21 Inhibition rate of test substances on oat peroxidase acyl-CoA oxidase (%)

[0241] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 2.28±0.42 5.43±0.66 14.04±0.56 13.02±0.73 3.55±0.44 20.61±0.12 15.04±0.77 6.68±0.26 Protein Peptides 11.21±0.51 11.43±0.05 3.44±0.02 7.66±1.07 13.36±0.11 10.07±0.04 5.00±1.10 0.92±0.43 polyphenols 19.22±0.04 17.92±0.08 14.16±0.02 15.49±0.07 14.13±0.09 13.73±0.15 0.76±0.34 3.05±0.06

[0242] Table 22 Inhibition rate of the test substances on quinoa peroxidase acyl-CoA oxidase (%)

[0243] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 3.39±0.42 12.13±0.07 7.91±0.32 2.40±0.03 13.28±0.82 22.00±0.06 10.47±0.85 12.69±0.36 Protein Peptides 5.38±0.25 10.35±0.03 8.02±0.50 3.87±0.95 8.95±0.04 10.52±0.48 8.33±0.50 13.97±0.36 polyphenols 16.56±0.86 18.01±0.07 14.16±0.02 2.34±0.25 2.62±0.01 9.12±0.05 3.04±0.41 11.64±0.72

[0244] Table 23 Inhibition rate of the test substances on buckwheat peroxidase acyl-CoA oxidase (%)

[0245] Grifola frondosa yellow umbrella Oyster mushrooms Nameko Oyster Mushroom Yellow elm mushroom Agrocybe tumefaciens Fungus Poria polysaccharides 8.63±0.55 11.54±0.04 12.67±0.01 3.20±0.76 13.78±0.02 18.79±0.08 5.13±0.05 8.31±0.07 Protein Peptides 8.80±0.05 4.91±0.72 2.41±0.47 6.63±0.04 10.83±0.05 8.10±0.04 1.88±0.01 4.09±0.05 polyphenols 20.42±0.05 19.03±0.06 3.84±0.09 6.22±0.79 11.22±0.66 12.29±0.54 16.31±0.06 6.10±0.07

[0246] Example 9: Detection Method for the Effect on Cereal Storage

[0247] During the storage of grains, lipase, lipoxygenase, peroxidase and peroxidase acyl-CoA oxidase synergistically catalyze lipid oxidation, leading to a significant accumulation of free fatty acids and an increase in oxidation products such as malondialdehyde.

[0248] The test grains were ground into powder using a grinder and passed through a 60-mesh sieve to obtain grain powder.

[0249] Test group 1 (expressed as 0.5%): 20g of cereal powder and 0.1g of the test substance were stirred and mixed evenly, placed in a sterile centrifuge tube, and placed in a constant temperature incubator at 60±1℃ for 14 days, and samples were taken every 48 hours.

[0250] Test group 2 (expressed as 1%): 20 g of cereal powder and 0.2 g of the test substance were stirred and mixed evenly, placed in a sterile centrifuge tube, and placed in a constant temperature incubator at 60±1°C for 14 days, with sampling every 48 hours.

[0251] Test group 3 (expressed as 1.5%): 20 g of cereal powder and 0.3 g of the test substance were stirred and mixed evenly, placed in a sterile centrifuge tube, and placed in a constant temperature incubator at 60±1°C for 14 days, with sampling every 48 hours.

[0252] Control group: 20 g of grain powder was placed in a sterile centrifuge tube and placed in a constant temperature incubator at 60±1℃ for 14 days, with samples taken every 48 hours.

[0253] The sampling samples of the experimental group are experimental samples, and the sampling samples of the control group are control samples.

[0254] To test the sample's lipase activity: Use a lipase (LPS) activity assay kit according to the manufacturer's instructions. To prepare the sample solution: Take 0.1 g of sample (test or control), add 1.0 mL of Reagent 1 (provided in the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C and 12,000 rpm for 15 minutes. Collect the supernatant, which is the sample solution. The result is the sample's LPS activity, expressed in U / g, meaning the number of units of lipase activity per gram of sample.

[0255] Lipoxygenase activity in samples: Use a lipoxygenase (LOX) activity assay kit according to the manufacturer's instructions. Sample preparation: Take 0.1 g of sample (test or control), add 1 mL of extract (provided with the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C, 12,000 rpm, for 20 minutes. Collect the supernatant, which is the sample solution. The result is the LOX activity of the sample, expressed in U / mg (i.e., the number of units of lipoxygenase activity per milligram of sample).

[0256] To test the peroxidase activity of a sample: Use a peroxidase (POD) activity assay kit according to the manufacturer's instructions. To prepare the sample solution: Take 0.1 g of sample (test or control), add 1 mL of the extract (provided with the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C, 12,000 rpm, for 10 minutes. Collect the supernatant, which is the sample solution. The result is the POD activity of the sample, expressed in U / mg, which represents the number of U of peroxidase activity per mg of sample.

[0257] To test the peroxidase acyl-CoA oxidase activity in samples, use a peroxidase acyl-CoA oxidase (A-COX / AOX) ELISA kit according to the manufacturer's instructions. Sample preparation: Take 0.1 g of sample (test or control), add 1 mL of normal saline, mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C, 3000 rpm, and collect the supernatant, which is the sample solution. The result is the peroxidase acyl-CoA oxidase content in the sample, expressed in ng / g, meaning the number of ng of peroxidase acyl-CoA oxidase per gram of sample.

[0258] To test the free fatty acid content in grains, use a free fatty acid (FFA) assay kit according to the manufacturer's instructions. Sample solution preparation: Take 0.1 g of sample (test or control), add 1.0 mL of extract (provided with the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Centrifuge at 4°C, 8,000 rpm, for 10 minutes. The supernatant is the sample solution. The result is the free fatty acid content of the sample, expressed in μmol / mg, meaning the number of μmol of free fatty acids per mg of sample.

[0259] To test for malondialdehyde (MDA) content in grains, use a malondialdehyde (MDA) content detection kit according to the instructions. Sample solution preparation: Take 0.1g of sample (test sample or control sample), add 1.0mL of extract (provided by the kit), mix thoroughly, and incubate in an ice-water bath for 2 hours. Then, centrifuge at 4°C and 12,000 rpm for 10 minutes. The supernatant is the sample solution. The result is the MDA content in the sample, expressed in nmol / g, i.e., the number of nmol of MDA per gram of sample.

[0260] Example 10: Comparison of the effects of various test substances on grain storage

[0261] The test substances were: Grifola frondosa polyphenol extract, Pholiota adiposa polyphenol extract, and Agrocybe oleracea polysaccharide extract, all of which were prepared in Example 4.

[0262] The tested grains were (all commercially available): millet, oats, quinoa or buckwheat.

[0263] Take the test substance and detect the effect of the test substance on the storage of grains according to the method of Example 9.

[0264] The lipase activity of millet samples collected at different sampling times is shown in Table 24. The lipase activity of oat samples collected at different sampling times is shown in Table 25. The lipase activity of quinoa samples collected at different sampling times is shown in Table 26. The lipase activity of buckwheat samples collected at different sampling times is shown in Table 27. During storage, the lipase activity of the grains showed an initial increase followed by a decrease (peaking between days 6 and 8). During the initial storage period, LPS activity of the grains increased significantly, likely due to their strong vitality and respiration capacity. External factors such as oxygen and humidity also contributed to the increase in LPS activity, accelerating lipid metabolism and decomposition. With extended storage, the free water content and substrate concentration in the grains decreased, leading to decreased LPS activity or enzyme inactivation at high temperatures. Compared with the control samples collected during the same period, the addition of the test substance significantly inhibited the lipase activity of the grains. This shows that the test substance can slow down the oxidation of grains, extend their storage period and maintain their nutritional value.

[0265] Table 24 Lipase activity (U / g) of samples collected at different sampling times when the coarse grains were millet

[0266] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 24.32±0.24 26.54±0.24 28.11±0.25 32.56±0.25 33.42±0.58 30.75±0.58 28.07±0.36 23.49±0.29 1% Maitake mushroom polyphenols 24.15±0.38 26.38±0.38 27.18±0.39 31.63±0.39 32.53±0.62 29.86±0.62 26.14±0.30 21.56±0.20 1.5% Maitake mushroom polyphenols 24.15±0.18 26.38±0.18 26.50±0.21 30.95±0.21 32.27±0.94 29.60±0.94 25.85±0.27 21.26±0.16 0.5% Phellodendron chinense polyphenols 24.64±0.31 26.87±0.31 27.88±0.45 32.33±0.45 33.45±0.36 30.78±0.36 28.35±0.54 23.77±0.52 1% Phellodendron chinense polyphenols 24.60±0.50 26.82±0.50 27.34±0.13 31.79±0.13 31.61±0.52 28.94±0.52 27.39±0.21 22.81±0.11 1.5% Phellodendron chinense polyphenols 24.05±0.25 26.28±0.25 27.06±0.56 31.51±0.56 31.08±0.14 28.41±0.14 27.16±0.51 22.58±0.46 0.5% Agrocybe edulis polysaccharide 24.35±0.19 26.57±0.19 28.63±0.32 33.08±0.32 33.21±0.34 30.55±0.34 27.76±0.54 23.18±0.41 1% Agrocybe edulis polysaccharide 24.29±0.51 26.51±0.51 28.54±0.48 32.99±0.48 32.38±0.08 29.72±0.08 27.30±0.32 22.72±0.21 1.5% Agrocybe edulis polysaccharide 24.23±0.19 26.45±0.19 28.23±0.17 32.68±0.17 31.82±0.09 29.15±0.09 26.78±0.65 22.20±0.51 Control samples 24.67±0.48 27.08±0.67 29.08±0.32 33.53±0.32 38.39±0.27 34.45±0.48 28.57±0.36 23.99±0.23

[0267] Table 25 Lipase activity of oats at different sampling times (U / g)

[0268] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 27.09±0.36 28.29±0.40 30.19±0.45 34.06±0.55 38.02±0.18 32.09±0.23 26.45±0.48 23.67±0.41 1% Maitake mushroom polyphenols 27.06±0.20 28.26±0.22 30.16±0.12 33.41±0.47 37.99±0.31 32.06±0.55 26.42±0.66 23.64±0.18 1.5% Maitake mushroom polyphenols 26.99±0.59 28.19±0.57 30.09±0.49 32.71±0.42 36.61±0.81 31.98±1.02 25.76±0.59 22.82±0.39 0.5% Phellodendron chinense polyphenols 27.15±0.39 28.35±0.40 30.25±0.29 33.93±0.33 38.72±0.37 32.78±0.39 27.15±0.31 23.67±0.36 1% Phellodendron chinense polyphenols 26.63±0.34 27.83±0.30 29.73±0.31 33.54±0.32 38.20±0.16 32.27±0.40 26.63±0.60 23.15±0.33 1.5% Phellodendron chinense polyphenols 26.29±0.31 27.49±0.32 29.39±0.21 32.99±0.40 36.97±0.61 31.92±0.32 26.29±0.31 22.81±0.29 0.5% Agrocybe edulis polysaccharide 27.55±0.41 28.75±0.45 30.65±0.48 33.23±0.40 39.99±0.13 34.06±0.16 28.43±0.31 23.15±0.14 1% Agrocybe edulis polysaccharide 27.52±0.49 28.56±0.54 30.04±0.30 32.56±0.31 39.00±0.39 32.95±0.48 27.98±0.20 22.24±0.42 1.5% Agrocybe edulis polysaccharide 27.46±0.33 28.66±0.32 30.56±0.25 32.31±0.11 37.74±0.48 31.66±0.22 27.21±0.30 22.08±0.03 Control samples 27.73±0.07 28.93±0.10 31.21±0.17 35.14±0.12 42.41±0.33 36.48±0.43 30.84±0.41 25.42±0.45

[0269] Table 26 Lipase activity (U / g) of samples collected at different sampling times when the coarse grain was quinoa

[0270] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 26.10±0.38 27.11±0.26 28.63±0.36 32.16±0.25 33.67±0.61 31.23±0.60 31.24±0.52 25.82±0.30 1% Maitake mushroom polyphenols 26.04±0.24 26.94±0.38 27.70±0.42 31.23±0.39 32.78±0.66 30.34±0.64 29.31±0.34 23.89±0.18 1.5% Maitake mushroom polyphenols 25.98±0.46 26.94±0.16 27.02±0.40 30.55±0.21 31.63±0.15 29.77±0.65 28.87±0.16 23.44±0.18 0.5% Phellodendron chinense polyphenols 26.07±0.29 27.43±0.33 28.29±0.59 31.72±0.43 33.70±0.30 31.26±0.34 31.52±0.70 26.10±0.53 1% Phellodendron chinense polyphenols 25.90±0.22 27.39±0.49 27.76±0.66 31.18±0.12 31.87±0.55 29.42±0.54 30.56±0.35 25.13±0.12 1.5% Phellodendron chinense polyphenols 25.90±0.52 26.84±0.26 27.48±0.35 29.80±0.04 30.78±0.57 28.88±0.14 29.15±0.32 23.86±0.22 0.5% Agrocybe edulis polysaccharide 26.39±0.38 27.14±0.20 29.15±0.78 32.68±0.32 33.47±0.40 31.02±0.36 30.93±0.66 25.50±0.39 1% Agrocybe edulis polysaccharide 26.35±0.22 27.08±0.53 29.06±0.17 32.59±0.48 32.64±0.14 30.19±0.10 30.47±0.46 25.04±0.20 1.5% Agrocybe edulis polysaccharide 25.80±0.28 27.02±0.17 28.75±0.48 32.28±0.17 31.63±0.04 29.18±0.08 29.06±0.33 23.64±0.07 Control samples 26.42±0.12 27.64±0.68 29.60±0.24 33.16±0.36 38.64±0.29 34.92±0.46 31.75±0.48 26.32±0.21

[0271] Table 27 Lipase activity (U / g) of samples collected at different sampling times when the coarse grain was buckwheat

[0272] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 24.84±0.22 27.45±0.29 28.34±0.19 33.85±0.29 33.35±0.38 30.31±0.35 26.82±0.63 22.67±0.38 1% Maitake mushroom polyphenols 24.78±0.51 27.34±0.36 28.17±0.34 33.76±0.50 32.52±0.12 29.42±0.07 26.36±0.44 22.21±0.19 1.5% Maitake mushroom polyphenols 24.69±0.36 27.34±0.18 27.97±0.21 32.00±0.63 31.87±0.14 28.91±0.09 24.36±0.05 20.80±0.05 0.5% Phellodendron chinense polyphenols 24.81±0.32 27.83±0.29 28.66±0.27 33.27±0.16 33.56±0.59 30.52±0.56 27.14±0.50 22.98±0.27 1% Phellodendron chinense polyphenols 24.64±0.51 27.79±0.48 28.62±0.46 32.40±0.38 32.67±0.64 29.63±0.60 25.21±0.32 21.06±0.20 1.5% Phellodendron chinense polyphenols 24.64±0.36 27.16±0.11 27.94±0.12 30.72±0.48 31.82±0.40 28.83±0.76 24.41±0.73 20.33±0.16 0.5% Agrocybe edulis polysaccharide 25.13±0.27 27.54±0.16 28.34±0.15 33.02±0.56 33.59±0.32 30.55±0.37 27.42±0.68 23.27±0.50 1% Agrocybe edulis polysaccharide 25.07±0.65 27.37±0.57 28.31±0.46 32.56±0.12 31.75±0.53 28.71±0.50 26.45±0.33 22.30±0.09 1.5% Agrocybe edulis polysaccharide 24.53±0.33 27.42±0.18 28.19±0.21 32.18±0.61 31.21±0.13 28.17±0.11 24.15±0.12 20.30±0.32 Control samples 25.16±0.52 28.04±0.64 29.00±0.62 34.30±0.32 38.52±0.27 34.21±0.46 27.64±0.45 23.49±0.20

[0273] The lipoxygenase activity of millet samples collected at different sampling times is shown in Table 28. The lipoxygenase activity of oats samples collected at different sampling times is shown in Table 29. The lipoxygenase activity of quinoa samples collected at different sampling times is shown in Table 30. The lipoxygenase activity of buckwheat samples collected at different sampling times is shown in Table 31. During storage, the lipoxygenase activity of the grains showed an initial decrease, followed by an increase, and then a subsequent decrease (peaking between days 6 and 10). Initially, natural antioxidants (such as phenolic compounds) present in the grains may inhibit LOX activity, or substrate concentrations may decrease with prolonged storage, leading to an increase in the concentration of LOX reaction products, causing LOX to enter a reversible inhibition state and a continuous decrease in LOX activity. Subsequently, LPS activity in the grains increased, further hydrolyzing triglycerides and releasing more FFAs, which provided more substrate for LOX and significantly increased LOX activity. Subsequently, LOX substrates (such as unsaturated fatty acids) may be gradually depleted due to the initial oxidation reaction, leading to decreased enzyme activity and loss of LOX activity due to self-inactivation or degradation by proteases. Compared with the control sample at the same time, the addition of the test substance significantly inhibited the activity of lipoxygenase in grains. This suggests that the test substance can slow the oxidation of grains, extend their storage life, and maintain their nutritional value.

[0274] Table 28 Lipoxygenase activity (U / mg) of samples collected at different sampling times when the coarse grains were millet

[0275]

[0276]

[0277] Table 29 Lipoxygenase activity of samples taken at different sampling times when the coarse grains were oats (U / mg)

[0278] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 2.17±0.17 1.94±0.19 1.89±0.10 2.39±0.35 2.44±0.25 2.00±0.33 1.94±0.19 1.67±0.17 1% Maitake mushroom polyphenols 2.17±0.00 1.94±0.10 1.83±0.00 2.22±0.25 1.83±0.00 1.83±0.33 1.83±0.00 1.56±0.10 1.5% Maitake mushroom polyphenols 2.28±0.19 2.06±0.10 1.94±0.19 1.89±0.10 2.00±0.44 1.50±0.00 1.44±0.19 1.17±0.17 0.5% Phellodendron chinense polyphenols 2.11±0.25 1.89±0.19 1.83±0.17 2.67±0.29 2.50±0.44 2.28±0.35 2.22±0.10 1.94±0.10 1% Phellodendron chinense polyphenols 2.22±0.19 2.00±0.17 1.89±0.19 2.56±0.10 2.33±0.50 2.17±0.17 2.12±0.10 1.84±0.01 1.5% Phellodendron chinense polyphenols 2.17±0.00 1.94±0.10 1.83±0.00 2.06±0.25 2.00±0.17 1.67±0.33 1.61±0.38 1.33±0.17 0.5% Agrocybe edulis polysaccharide 2.22±0.10 2.00±0.00 1.89±0.10 2.33±0.29 2.78±0.51 1.94±0.38 1.89±0.35 1.61±0.25 1% Agrocybe edulis polysaccharide 2.17±0.17 1.94±0.19 1.78±0.10 1.78±0.10 2.12±0.20 1.56±0.10 1.61±0.10 1.56±0.19 1.5% Agrocybe edulis polysaccharide 2.28±0.10 2.06±0.10 1.94±0.10 1.89±0.19 2.17±0.17 1.50±0.29 1.61±0.19 1.33±0.17 Control samples 2.28±0.10 2.06±0.10 1.94±0.10 2.89±0.19 3.00±0.29 2.50±0.17 2.44±0.10 2.17±0.17

[0279] Table 30 Lipoxygenase activity (U / mg) of samples taken at different sampling times when the coarse grain was quinoa

[0280] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 2.00±0.29 1.83±0.00 1.67±0.17 1.83±0.50 2.44±0.35 2.33±0.44 1.89±0.42 1.61±0.25 1% Maitake mushroom polyphenols 2.00±0.17 1.78±0.10 1.61±0.10 1.78±0.54 2.56±0.63 2.17±0.29 1.72±0.10 1.50±0.17 1.5% Maitake mushroom polyphenols 1.94±0.10 1.78±0.10 1.61±0.10 1.89±0.38 1.72±0.48 1.83±0.17 1.61±0.35 1.11±0.10 0.5% Phellodendron chinense polyphenols 2.00±0.17 1.89±0.10 1.61±0.10 2.06±0.42 2.72±0.42 2.61±0.35 2.22±0.25 1.89±0.10 1% Phellodendron chinense polyphenols 2.00±0.17 1.83±0.00 1.67±0.17 1.99±0.29 2.50±0.44 2.50±0.17 2.12±0.20 1.78±0.09 1.5% Phellodendron chinense polyphenols 1.94±0.25 1.72±0.10 1.56±0.19 1.89±0.10 1.89±0.10 1.72±0.25 1.61±0.42 1.28±0.19 0.5% Agrocybe edulis polysaccharide 2.00±0.00 1.83±0.17 1.72±0.10 1.94±0.48 2.22±0.59 2.28±0.25 1.94±0.10 1.56±0.19 1% Agrocybe edulis polysaccharide 1.94±0.10 1.83±0.17 1.72±0.19 1.78±0.75 2.33±0.33 2.22±0.10 1.83±0.17 1.39±0.19 1.5% Agrocybe edulis polysaccharide 1.94±0.10 1.78±0.10 1.67±0.17 1.83±0.17 1.94±0.38 1.83±0.17 1.44±0.35 1.28±0.10 Control samples 2.00±0.00 1.89±0.10 1.78±0.10 2.11±0.35 2.94±0.19 2.83±0.29 2.44±0.19 2.11±0.19

[0281] Table 31 Lipoxygenase activity (U / mg) of samples collected at different sampling times when the coarse grain was buckwheat

[0282] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 1.67±0.00 1.56±0.10 1.33±0.17 1.83±0.17 2.00±0.29 2.17±0.00 1.72±0.10 1.33±0.17 1% Maitake mushroom polyphenols 1.72±0.35 1.50±0.17 1.28±0.19 1.72±0.35 1.94±0.10 2.17±0.29 1.67±0.17 1.17±0.17 1.5% Maitake mushroom polyphenols 1.67±0.17 1.50±0.17 1.28±0.10 1.61±0.10 1.89±0.51 1.83±0.33 1.56±0.10 1.11±0.10 0.5% Phellodendron chinense polyphenols 1.78±0.25 1.61±0.10 1.28±0.10 1.89±0.35 2.00±0.17 2.11±0.19 1.67±0.17 1.22±0.10 1% Phellodendron chinense polyphenols 1.78±0.25 1.56±0.10 1.33±0.17 1.78±0.25 1.94±0.35 2.11±0.19 1.67±0.33 1.17±0.17 1.5% Phellodendron chinense polyphenols 1.72±0.10 1.44±0.19 1.28±0.25 1.61±0.10 1.67±0.17 1.72±0.19 1.56±0.25 1.06±0.10 0.5% Agrocybe edulis polysaccharide 1.78±0.10 1.56±0.19 1.39±0.10 1.78±0.35 2.00±0.33 2.11±0.19 1.67±0.33 1.39±0.19 1% Agrocybe edulis polysaccharide 1.72±0.19 1.56±0.25 1.39±0.19 1.72±0.19 1.89±0.25 2.11±0.10 1.61±0.35 1.33±0.00 1.5% Agrocybe edulis polysaccharide 1.72±0.19 1.50±0.17 1.33±0.17 1.67±0.00 1.89±0.35 2.00±0.29 1.56±0.10 1.17±0.00 Control samples 1.78±0.10 1.61±0.10 1.44±0.10 1.94±0.19 2.50±0.29 2.28±0.10 1.78±0.10 1.44±0.10

[0283] The peroxidase activity of millet samples collected at different sampling times is shown in Table 32. The peroxidase activity of oat samples collected at different sampling times is shown in Table 33. The peroxidase activity of quinoa samples collected at different sampling times is shown in Table 34. The peroxidase activity of buckwheat samples collected at different sampling times is shown in Table 35. During storage, the peroxidase activity of the grains showed an initial upward and then downward trend. During the initial storage of grains, the lipid peroxidation reaction rate is rapid, resulting in relatively high peroxide production and an increase in POD. With extended storage, the substrate concentration decreases, the reaction rate slows, the amount of peroxide produced decreases, and POD activity decreases. Compared with the control samples collected during the same period, the addition of the test substance significantly inhibited the peroxidase activity of the grains. This suggests that the test substance can slow the oxidation of the grains, extend their storage life, and maintain their nutritional value.

[0284] Table 32 Peroxidase activity of samples taken at different sampling times when the coarse grains were millet (U / mg)

[0285] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 6.04±0.16 6.39±0.43 7.38±0.13 8.77±0.07 10.30±0.21 10.02±0.03 8.36±0.18 7.87±0.11 1% Maitake mushroom polyphenols 6.03±0.16 6.38±0.35 7.22±0.12 8.86±0.11 9.95±0.11 9.25±0.09 8.20±0.16 7.61±0.04 1.5% Maitake mushroom polyphenols 6.04±0.16 6.36±0.22 7.05±0.04 8.16±0.11 9.14±0.14 8.65±0.08 8.00±0.12 7.41±0.08 0.5% Phellodendron chinense polyphenols 6.04±0.16 6.35±0.22 7.21±0.00 8.90±0.21 10.01±0.11 9.64±0.05 8.08±0.09 7.66±0.09 1% Phellodendron chinense polyphenols 6.04±0.16 6.50±0.31 7.49±0.03 9.18±0.12 10.49±0.21 9.89±0.05 8.47±0.13 7.75±0.06 1.5% Phellodendron chinense polyphenols 6.04±0.13 6.47±0.24 7.02±0.11 8.27±0.10 9.12±0.08 8.70±0.05 8.00±0.10 7.13±0.04 0.5% Agrocybe edulis polysaccharide 5.99±0.34 6.43±0.36 7.42±0.04 8.89±0.27 10.09±0.21 9.59±0.06 8.40±0.14 7.81±0.08 1% Agrocybe edulis polysaccharide 6.05±0.16 6.54±0.44 7.09±0.02 8.24±0.20 9.04±0.07 9.01±0.02 8.08±0.07 7.60±0.12 1.5% Agrocybe edulis polysaccharide 6.08±0.42 6.04±0.27 7.00±0.05 7.70±0.06 9.04±0.06 8.88±0.04 8.09±0.09 7.06±0.10 Control samples 6.05±0.14 6.62±0.03 7.61±0.04 9.41±0.31 10.93±0.15 10.55±0.16 8.81±0.11 8.22±0.08

[0286] Table 33 Peroxidase activity of samples taken at different sampling times when the coarse grains were oats (U / mg)

[0287] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 0.35±0.02 0.43±0.02 0.54±0.03 0.73±0.02 0.75±0.00 0.60±0.01 0.49±0.01 0.43±0.00 1% Maitake mushroom polyphenols 0.35±0.02 0.41±0.01 0.50±0.04 0.68±0.01 0.73±0.01 0.58±0.01 0.56±0.01 0.49±0.01 1.5% Maitake mushroom polyphenols 0.35±0.01 0.43±0.01 0.48±0.03 0.65±0.00 0.69±0.00 0.55±0.01 0.51±0.02 0.42±0.01 0.5% Phellodendron chinense polyphenols 0.36±0.01 0.44±0.02 0.54±0.02 0.71±0.01 0.71±0.00 0.66±0.01 0.56±0.01 0.48±0.02 1% Phellodendron chinense polyphenols 0.38±0.04 0.44±0.04 0.45±0.06 0.64±0.01 0.68±0.02 0.62±0.02 0.51±0.02 0.43±0.02 1.5% Phellodendron chinense polyphenols 0.38±0.02 0.43±0.04 0.51±0.00 0.61±0.01 0.66±0.01 0.61±0.00 0.50±0.01 0.40±0.01 0.5% Agrocybe edulis polysaccharide 0.37±0.04 0.45±0.04 0.55±0.02 0.74±0.01 0.71±0.01 0.69±0.02 0.58±0.02 0.50±0.01 1% Agrocybe edulis polysaccharide 0.34±0.01 0.44±0.03 0.48±0.03 0.65±0.01 0.69±0.01 0.64±0.01 0.52±0.00 0.47±0.01 1.5% Agrocybe edulis polysaccharide 0.36±0.03 0.43±0.01 0.47±0.00 0.64±0.00 0.68±0.00 0.61±0.00 0.51±0.00 0.41±0.01 Control samples 0.37±0.07 0.45±0.06 0.56±0.01 0.75±0.01 0.80±0.00 0.74±0.00 0.64±0.02 0.56±0.04

[0288] Table 34 Peroxidase activity of samples taken at different sampling times when the coarse grain was quinoa (U / mg)

[0289] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 5.90±0.32 6.03±0.31 6.29±0.17 7.12±0.15 8.93±0.16 7.12±0.16 6.56±0.18 6.20±0.12 1% Maitake mushroom polyphenols 5.89±0.32 6.03±0.31 6.11±0.10 7.11±0.19 8.59±0.10 6.45±0.03 6.40±0.16 6.04±0.09 1.5% Maitake mushroom polyphenols 5.89±0.32 6.03±0.32 5.79±0.19 6.72±0.06 7.88±0.20 6.18±0.18 6.16±0.04 5.73±0.07 0.5% Phellodendron chinense polyphenols 5.90±0.32 6.03±0.31 6.02±0.10 7.24±0.14 8.64±0.11 6.83±0.14 6.29±0.08 5.92±0.11 1% Phellodendron chinense polyphenols 5.88±0.34 6.02±0.34 6.20±0.14 7.09±0.23 8.90±0.21 7.09±0.53 6.13±0.05 5.77±0.03 1.5% Phellodendron chinense polyphenols 5.89±0.28 6.03±0.27 5.76±0.08 6.72±0.19 8.10±0.13 5.95±0.09 5.85±0.15 5.59±0.17 0.5% Agrocybe edulis polysaccharide 5.84±0.50 5.98±0.49 6.17±0.13 7.23±0.21 8.72±0.16 6.80±0.06 6.60±0.06 6.24±0.01 1% Agrocybe edulis polysaccharide 5.90±0.32 6.04±0.31 5.70±0.11 6.04±0.14 7.67±0.16 5.85±0.10 6.17±0.12 5.81±0.18 1.5% Agrocybe edulis polysaccharide 5.93±0.56 6.07±0.55 5.61±0.06 5.99±0.13 7.65±0.13 5.72±0.14 5.64±0.16 5.58±0.15 Control samples 5.97±0.19 6.11±0.18 6.53±0.07 7.75±0.13 9.57±0.08 7.75±0.05 6.90±0.09 6.54±0.06

[0290] Table 35 Peroxidase activity of samples taken at different sampling times when the coarse grain was buckwheat (U / mg)

[0291] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 0.57±0.01 0.62±0.01 0.63±0.01 0.67±0.01 1.08±0.02 0.66±0.02 0.63±0.03 0.60±0.01 1% Maitake mushroom polyphenols 0.57±0.02 0.61±0.00 0.62±0.01 0.66±0.05 1.07±0.03 0.61±0.02 0.59±0.03 0.55±0.00 1.5% Maitake mushroom polyphenols 0.57±0.01 0.60±0.01 0.59±0.01 0.62±0.01 1.07±0.04 0.61±0.00 0.58±0.01 0.54±0.01 0.5% Phellodendron chinense polyphenols 0.56±0.02 0.61±0.00 0.67±0.01 0.66±0.02 1.05±0.06 0.61±0.03 0.59±0.01 0.56±0.00 1% Phellodendron chinense polyphenols 0.57±0.01 0.62±0.01 0.60±0.04 0.65±0.02 1.08±0.02 0.60±0.01 0.58±0.01 0.55±0.01 1.5% Phellodendron chinense polyphenols 0.57±0.00 0.62±0.02 0.59±0.00 0.61±0.01 1.07±0.04 0.56±0.00 0.54±0.01 0.51±0.01 0.5% Agrocybe edulis polysaccharide 0.54±0.03 0.59±0.02 0.65±0.01 0.66±0.01 1.02±0.10 0.63±0.01 0.60±0.01 0.57±0.02 1% Agrocybe edulis polysaccharide 0.59±0.00 0.64±0.03 0.67±0.01 0.66±0.00 1.12±0.06 0.62±0.01 0.59±0.01 0.56±0.02 1.5% Agrocybe edulis polysaccharide 0.57±0.00 0.60±0.01 0.63±0.00 0.65±0.01 1.08±0.04 0.57±0.01 0.56±0.01 0.52±0.01 Control samples 0.59±0.01 0.63±0.01 0.72±0.01 0.73±0.01 1.11±0.02 0.69±0.01 0.67±0.00 0.63±0.04

[0292] The peroxidase acyl-CoA oxidase activities of millet samples collected at different sampling times are shown in Table 36. The peroxidase acyl-CoA oxidase activities of oats samples collected at different sampling times are shown in Table 37. The peroxidase acyl-CoA oxidase activities of quinoa samples collected at different sampling times are shown in Table 38. The peroxidase acyl-CoA oxidase activities of buckwheat samples collected at different sampling times are shown in Table 39. During storage, the peroxidase acyl-CoA oxidase activities of the grains showed a trend of first increasing and then decreasing (peaking on days 4-8). During the initial storage of grains, A-COX activity is low, as fatty acid oxidation has not yet fully initiated, and antioxidants in the grains may inhibit its activity. As storage progresses, fatty acids in the grains begin to oxidize, and A-COX activity gradually increases. This accelerated oxidation of fatty acids activates A-COX, a key enzyme, to meet metabolic needs. Depletion of fatty acid substrates leads to decreased enzyme activity. Compared with the control sample, the addition of the test substance significantly inhibited the activity of the peroxidase acyl-CoA oxidase in the grains. This suggests that the test substance can slow oxidation of the grains, extend their storage life, and maintain their nutritional value.

[0293] Table 36 Peroxidase acyl-CoA oxidase activity of samples collected at different sampling times when the coarse grains were millet (ng / g)

[0294]

[0295] Table 37 Peroxidase Acyl-CoA Oxidase Activity of Samples Taken at Different Sampling Times When the Cereals are Oats (ng / g)

[0296]

[0297] Table 38 Peroxidase acyl-CoA oxidase activity of samples collected at different sampling times when the coarse grain was quinoa (ng / g)

[0298]

[0299]

[0300] Table 39 Peroxidase acyl-CoA oxidase activity of samples collected at different sampling times when the coarse grain was buckwheat (ng / g)

[0301]

[0302] The free fatty acid content of millet samples collected at different sampling times is shown in Table 40. The free fatty acid content of oat samples collected at different sampling times is shown in Table 41. The free fatty acid content of quinoa samples collected at different sampling times is shown in Table 42. The free fatty acid content of buckwheat samples collected at different sampling times is shown in Table 43. The free fatty acid content initially increased and then decreased with storage time (peaking on the eighth day). During the initial storage period, FFA content was relatively low, primarily in the form of triglycerides. LPS activity was not yet fully activated, resulting in low FFA production. With extended storage, LPS activity gradually increased, and triglycerides were hydrolyzed into free fatty acids and glycerol, leading to a significant increase in FFA content. Unsaturated fatty acids in the grains were more susceptible to oxidation, further promoting FFA accumulation. In the later stages of storage, FFAs were further decomposed into small molecules such as aldehydes, ketones, and acids during oxidation, resulting in a decrease in their content. Compared with the control sample of the same period, the addition of the test substance significantly reduced the free fatty acid content, which indicates that the test substance helps maintain the quality of grains.

[0303] Table 40 Free fatty acid content of samples taken at different sampling times when the coarse grains were millet (μmol / mg)

[0304]

[0305]

[0306] Table 41 Free fatty acid content of oats in samples taken at different sampling times (μmol / mg)

[0307] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 2.56±0.05 2.82±0.04 2.81±0.09 3.18±0.08 3.66±0.09 3.30±0.11 3.03±0.09 2.70±0.08 1% Maitake mushroom polyphenols 2.56±0.05 2.82±0.04 2.81±0.09 3.17±0.09 3.66±0.09 3.30±0.11 3.03±0.09 2.66±0.12 1.5% Maitake mushroom polyphenols 2.56±0.05 2.81±0.04 2.60±0.11 2.97±0.11 3.44±0.10 3.09±0.08 2.80±0.09 2.49±0.20 0.5% Phellodendron chinense polyphenols 2.60±0.03 2.48±0.02 2.58±0.10 2.95±0.09 3.43±0.09 3.07±0.11 2.80±0.09 2.50±0.04 1% Phellodendron chinense polyphenols 2.60±0.03 2.48±0.02 2.58±0.10 2.95±0.09 3.43±0.10 3.05±0.10 2.79±0.09 2.49±0.05 1.5% Phellodendron chinense polyphenols 2.60±0.03 2.48±0.02 2.58±0.10 2.94±0.10 3.42±0.10 3.03±0.10 2.80±0.10 2.49±0.05 0.5% Agrocybe edulis polysaccharide 2.57±0.04 2.71±0.03 2.79±0.10 3.15±0.09 3.64±0.10 3.27±0.14 3.01±0.10 2.69±0.07 1% Agrocybe edulis polysaccharide 2.57±0.03 2.70±0.04 2.78±0.11 3.06±0.08 3.57±0.08 3.20±0.05 2.66±0.17 2.51±0.02 1.5% Agrocybe edulis polysaccharide 2.57±0.04 2.71±0.03 2.79±0.10 2.94±0.01 3.47±0.01 3.09±0.02 2.66±0.16 2.48±0.06 Control samples 2.64±0.29 2.85±0.12 2.96±0.08 3.40±0.02 3.89±0.03 3.53±0.05 3.26±0.03 2.96±0.13

[0308] Table 42 Free fatty acid content of samples taken at different sampling times when quinoa is used as coarse grain (μmol / mg)

[0309] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 2.14±0.04 2.31±0.04 2.43±0.09 2.80±0.09 3.10±0.10 2.99±0.12 2.84±0.09 2.59±0.08 1% Maitake mushroom polyphenols 2.13±0.04 2.31±0.04 2.43±0.10 2.80±0.09 3.11±0.11 2.99±0.12 2.84±0.09 2.55±0.12 1.5% Maitake mushroom polyphenols 2.13±0.04 2.31±0.04 2.25±0.16 2.59±0.11 3.10±0.11 2.69±0.01 2.55±0.03 2.38±0.20 0.5% Phellodendron chinense polyphenols 2.14±0.03 2.21±0.02 2.20±0.10 2.56±0.09 3.34±0.10 2.76±0.12 2.61±0.09 2.38±0.06 1% Phellodendron chinense polyphenols 2.14±0.03 2.20±0.01 2.20±0.10 2.57±0.10 3.33±0.10 2.74±0.10 2.60±0.09 2.38±0.05 1.5% Phellodendron chinense polyphenols 2.14±0.03 2.20±0.01 2.16±0.13 2.56±0.11 3.02±0.06 2.72±0.10 2.59±0.02 2.38±0.05 0.5% Agrocybe edulis polysaccharide 2.15±0.03 2.21±0.03 2.40±0.10 2.77±0.11 3.28±0.07 2.95±0.14 2.82±0.10 2.59±0.05 1% Agrocybe edulis polysaccharide 2.15±0.03 2.20±0.03 2.39±0.12 2.78±0.10 3.29±0.08 2.95±0.13 2.82±0.10 2.57±0.03 1.5% Agrocybe edulis polysaccharide 2.15±0.03 2.20±0.03 2.39±0.10 2.72±0.05 3.06±0.01 2.72±0.01 2.54±0.02 2.40±0.01 Control samples 2.22±0.29 2.35±0.12 2.58±0.09 3.03±0.02 3.57±0.04 3.21±0.06 3.07±0.03 2.85±0.13

[0310] Table 43 Free fatty acid content of samples taken at different sampling times when the coarse grain is buckwheat (μmol / mg)

[0311] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 1.69±0.04 1.85±0.04 1.89±0.07 2.41±0.08 2.82±0.09 2.53±0.11 2.18±0.09 2.06±0.08 1% Maitake mushroom polyphenols 1.69±0.04 1.84±0.04 1.88±0.07 2.40±0.09 2.82±0.09 2.53±0.12 2.18±0.09 2.02±0.12 1.5% Maitake mushroom polyphenols 1.69±0.04 1.84±0.04 1.68±0.13 2.16±0.06 2.56±0.03 2.28±0.02 1.91±0.03 1.83±0.02 0.5% Phellodendron chinense polyphenols 1.47±0.02 1.51±0.02 1.66±0.08 2.18±0.09 2.59±0.09 2.30±0.12 1.95±0.09 1.86±0.04 1% Phellodendron chinense polyphenols 1.47±0.02 1.51±0.02 1.65±0.08 2.17±0.09 2.59±0.10 2.28±0.10 1.94±0.09 1.84±0.05 1.5% Phellodendron chinense polyphenols 1.47±0.02 1.51±0.02 1.66±0.08 2.16±0.09 2.57±0.09 2.26±0.10 1.91±0.03 1.84±0.01 0.5% Agrocybe edulis polysaccharide 1.70±0.03 1.74±0.03 1.87±0.07 2.38±0.09 2.80±0.09 2.50±0.14 2.16±0.10 2.06±0.06 1% Agrocybe edulis polysaccharide 1.70±0.03 1.74±0.03 1.86±0.08 2.38±0.09 2.80±0.10 2.50±0.12 2.15±0.10 2.04±0.02 1.5% Agrocybe edulis polysaccharide 1.70±0.03 1.74±0.03 1.86±0.08 2.18±0.01 2.50±0.00 2.29±0.03 1.90±0.01 1.84±0.01 Control samples 1.77±0.28 1.88±0.12 2.04±0.07 2.63±0.01 3.05±0.03 2.76±0.06 2.40±0.03 2.31±0.13

[0312] The malondialdehyde content of millet samples collected at different sampling times is shown in Table 44. The malondialdehyde content of oat samples collected at different sampling times is shown in Table 45. The malondialdehyde content of quinoa samples collected at different sampling times is shown in Table 46. The malondialdehyde content of buckwheat samples collected at different sampling times is shown in Table 47. Malondialdehyde content increases with storage time. With extended storage, lipids in the grains begin to oxidize, and FFA, under the action of LOX, forms hydroperoxides, which are further broken down into small molecules such as MDA. Compared with the control samples collected during the same period, the addition of the test substance significantly reduced the malondialdehyde content. This indicates that the test substance can effectively inhibit fat oxidation, reduce the formation of harmful substances such as MDA, and thus protect the quality of the grains.

[0313] Table 44 Malondialdehyde content (nmol / g) of samples taken at different sampling times when the coarse grains were millet

[0314]

[0315]

[0316] Table 45 Malondialdehyde content of samples taken at different sampling times when the coarse grains are oats (nmol / g)

[0317] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 9.73±0.25 10.55±0.87 12.08±0.70 17.38±0.70 17.71±0.22 20.70±1.04 21.92±0.81 23.62±0.94 1% Maitake mushroom polyphenols 9.66±0.17 10.41±0.81 11.97±0.50 17.20±0.88 17.44±0.32 20.52±1.03 21.74±0.64 23.39±0.70 1.5% Maitake mushroom polyphenols 9.71±0.30 10.41±0.41 12.11±0.84 15.82±0.53 16.70±0.32 18.87±0.28 20.11±0.65 21.13±0.05 0.5% Phellodendron chinense polyphenols 9.73±0.25 11.20±0.60 12.29±0.49 17.35±0.34 19.12±0.87 20.66±0.76 21.88±0.70 23.53±0.73 1% Phellodendron chinense polyphenols 9.73±0.35 11.21±0.78 11.97±0.62 16.68±1.32 18.26±0.37 19.80±0.62 21.02±1.01 22.67±0.95 1.5% Phellodendron chinense polyphenols 9.71±0.31 11.20±0.87 12.01±0.75 15.93±0.36 16.52±0.30 18.60±0.14 19.82±0.31 20.86±0.43 0.5% Agrocybe edulis polysaccharide 9.75±0.28 10.41±0.86 12.06±0.43 17.78±0.91 18.33±0.89 20.68±0.90 21.90±0.59 23.55±0.65 1% Agrocybe edulis polysaccharide 9.53±0.42 10.48±0.88 11.47±0.28 16.20±0.54 17.13±0.59 17.56±0.36 19.14±0.34 21.88±0.55 1.5% Agrocybe edulis polysaccharide 9.61±0.40 10.39±0.68 12.01±0.77 15.88±0.46 16.31±0.08 17.53±0.30 19.77±0.27 21.08±0.19 Control samples 9.73±0.30 11.72±0.42 12.85±0.55 19.50±0.08 21.51±0.14 23.37±0.20 24.32±0.11 25.75±0.09

[0318] Table 46 Malondialdehyde content of samples taken at different sampling times when quinoa is used as coarse grain (nmol / g)

[0319] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 10.75±0.28 11.88±0.53 12.16±0.65 17.19±1.16 17.90±0.28 20.84±1.01 22.31±0.78 24.18±0.97 1% Maitake mushroom polyphenols 10.68±0.19 11.77±0.41 12.02±0.28 17.01±1.29 17.63±0.33 20.66±1.00 22.13±0.62 23.94±0.73 1.5% Maitake mushroom polyphenols 10.73±0.32 11.92±0.64 12.02±0.17 16.20±1.18 17.17±0.24 19.01±0.27 20.07±0.44 21.31±0.36 0.5% Phellodendron chinense polyphenols 10.75±0.27 12.10±0.30 12.81±0.30 17.15±0.80 19.32±0.81 20.81±0.73 22.28±0.67 24.09±0.75 1% Phellodendron chinense polyphenols 10.75±0.39 11.77±0.34 12.83±0.50 16.49±1.02 18.46±0.34 19.95±0.65 21.42±1.02 23.23±0.92 1.5% Phellodendron chinense polyphenols 10.73±0.35 11.81±0.81 12.81±0.33 16.31±0.53 17.28±0.44 19.09±0.62 20.14±0.34 22.06±0.87 0.5% Agrocybe edulis polysaccharide 10.77±0.32 11.86±0.24 12.02±1.36 17.58±1.37 19.34±0.76 20.82±0.87 22.29±0.57 24.10±0.68 1% Agrocybe edulis polysaccharide 10.56±0.47 11.27±0.57 12.09±1.35 16.00±0.78 17.96±0.43 19.44±0.75 20.91±0.33 22.44±0.57 1.5% Agrocybe edulis polysaccharide 10.63±0.44 11.81±0.73 12.00±1.17 16.09±0.22 17.54±0.11 19.25±0.19 20.54±0.39 21.63±0.32 Control samples 10.75±0.34 12.65±0.36 13.33±0.56 19.30±0.47 21.70±0.16 23.51±0.17 24.71±0.12 26.31±0.06

[0320] Table 47 Malondialdehyde content of samples taken at different sampling times when the coarse grain is buckwheat (nmol / g)

[0321] Day 0 Day 2 Day 4 Day 6 Day 8 Day 10 Day 12 Day 14 0.5% Maitake mushroom polyphenols 10.97±0.05 11.97±0.08 13.32±0.49 19.39±0.91 21.20±0.22 23.44±0.98 24.71±0.86 25.21±0.91 1% Maitake mushroom polyphenols 10.95±0.08 11.86±0.33 13.21±0.45 19.21±0.93 20.93±0.32 23.26±1.01 24.53±0.73 24.98±0.67 1.5% Maitake mushroom polyphenols 10.95±0.06 11.79±0.35 13.35±0.64 18.40±1.45 20.48±0.81 21.20±0.32 22.47±0.32 22.90±0.34 0.5% Phellodendron chinense polyphenols 10.97±0.11 12.59±0.09 13.53±0.32 19.35±0.62 22.62±0.87 23.40±0.68 24.68±0.70 25.13±0.71 1% Phellodendron chinense polyphenols 10.97±0.05 12.59±0.14 13.15±0.50 18.69±1.00 21.76±0.37 22.54±0.68 23.82±0.90 24.27±0.98 1.5% Phellodendron chinense polyphenols 10.95±0.03 12.59±0.09 13.12±0.95 18.44±0.56 20.68±0.12 20.97±0.79 22.72±0.27 23.53±1.28 0.5% Agrocybe edulis polysaccharide 10.97±0.00 11.91±0.09 13.30±0.28 19.78±1.12 22.63±0.78 23.42±0.86 24.70±0.66 25.14±0.62 1% Agrocybe edulis polysaccharide 10.95±0.06 11.86±0.14 12.71±0.66 17.99±0.44 21.00±0.62 22.04±0.80 23.32±0.42 23.48±0.52 1.5% Agrocybe edulis polysaccharide 10.95±0.03 11.88±0.06 13.24±0.72 19.07±1.48 20.88±0.08 20.95±0.08 22.22±0.35 22.67±0.17 Control samples 10.97±0.00 12.24±0.09 14.09±0.32 21.51±0.42 25.00±0.14 26.11±0.16 27.11±0.22 27.35±0.12

[0322] The results of this example show that 1.5% maitake mushroom polyphenol extract or 1.5% phellodendron polyphenol extract or 1.5% Agrocybe aegerita polysaccharide extract has the most significant inhibitory effect on the activities of key oxidases (LPS, LOX, POD and A-COX) and secondary oxidation products (FFA and MDA) during the storage of grains.

[0323] Comparative Example

[0324] A parallel experiment was conducted using tea polyphenols as a positive control for the test substance in Example 6. Hydroxyl radical scavenging rate (%): 68.44 ± 0.28, superoxide anion radical scavenging rate (%): 30.64 ± 0.42, and ABTS radical scavenging rate (%): 57.14 ± 0.09.

[0325] Tea polyphenols were used as a positive control for the test substances in Example 8, and parallel tests were conducted. The inhibition rate of the test substance on millet lipase (%) was 31.51±0.37. The inhibition rate of the test substance on oat lipase (%) was 49.48±0.38. The inhibition rate of the test substance on quinoa lipase (%) was 35.74±0.02. The inhibition rate of the test substance on buckwheat lipase (%) was 28.34±0.15. The inhibition rate of the test substance on millet lipoxygenase (%) was 59.37±0.00. The inhibition rate of the test substance on oat lipoxygenase (%) was 68.18±3.93. The inhibition rate of the test substance on quinoa lipoxygenase (%) was 52.17±0.00. The inhibition rate of the test substance on buckwheat lipoxygenase (%) was 51.16±4.03. The inhibition rate of the test substance on millet polyphenol oxidase (%): 65.08±5.19. The inhibition rate of the test substance on oat polyphenol oxidase (%): 65.81±2.77. The inhibition rate of the test substance on quinoa polyphenol oxidase (%): 48.56±1.41. The inhibition rate of the test substance on buckwheat polyphenol oxidase (%): 60.30±8.33. The inhibition rate of the test substance on millet peroxidase (%): 29.68±0.51. The inhibition rate of the test substance on oat peroxidase (%): 32.00±0.94. The inhibition rate of the test substance on quinoa peroxidase (%): 33.76±0.32. The inhibition rate of the test substance on buckwheat peroxidase (%): 28.01±0.75. The inhibition rate of the test substance on millet peroxidase acyl-CoA oxidase (%) was 31.72±0.42. The inhibition rate of the test substance on oat peroxidase acyl-CoA oxidase (%) was 27.44±0.26. The inhibition rate of the test substance on quinoa peroxidase acyl-CoA oxidase (%) was 22.73±0.90. The inhibition rate of the test substance on buckwheat peroxidase acyl-CoA oxidase (%) was 21.62±0.44.

[0326] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. Application of edible fungus extracts; the application is as follows (I) or (II) or (III) or (IV) or (V): (I) Application of edible fungus extracts as antioxidants; (II) Application of edible fungus extracts in the preparation of antioxidants; (III) Application of edible fungus extracts in the preparation of additives for grain storage; (IV) Application of edible fungus extracts as additives in grain storage; (V) Application of edible fungus extracts as additives with antioxidant function in grain storage; The edible fungus extract is a polysaccharide extract of Agrocybe tumefaciens, a polyphenol extract of Pholiota adiposa, or a polyphenol extract of Grifola frondosa.

2. An antioxidant comprising an edible fungus extract; the edible fungus extract is a tea tree agaric polysaccharide extract, a phellodendron polyphenol extract, or a maitake mushroom polyphenol extract.

3. An additive for storing grains, comprising an edible fungus extract; the edible fungus extract is a tea tree agaric polysaccharide extract, a pholiota cap polyphenol extract, or a maitake mushroom polyphenol extract.

4. The use according to claim 1 or the antioxidant according to claim 2 or the additive according to claim 3, characterized in that: The antioxidant has the following functions: scavenging hydroxyl radicals and / or scavenging superoxide anion radicals and / or scavenging ABTS radicals; during storage of cereals, the additive plays the following function: inhibiting the accumulation of free fatty acids and / or malondialdehyde in the cereals.

5. The use according to claim 1 or the antioxidant according to claim 2 or the additive according to claim 3, characterized in that: The Agrocybe tumefaciens polysaccharide extract is prepared by using Agrocybe tumefaciens as the raw material through a water extraction-ethanol precipitation method; the Pholiota adiposa polyphenol extract is prepared by using Pholiota adiposa as the raw material through an ethanol aqueous solution extraction method; and the Grifola frondosa polyphenol extract is prepared by using Grifola frondosa as the raw material through an ethanol aqueous solution extraction method.

6. The use according to claim 1 or the antioxidant according to claim 2 or the additive according to claim 3, characterized in that: The Agrocybe fasciata polysaccharide extract is the Agrocybe fasciata polysaccharide extract described in claim 7; the Phellodendron umbellifer polyphenol extract is the Phellodendron umbellifer polyphenol extract described in claim 7; and the Grifola frondosa polyphenol extract is the Grifola frondosa polyphenol extract described in claim 7.

7. Edible fungus extract, characterized in that: The edible fungus extract is a polysaccharide extract of Agrocybe tumefaciens, a polyphenol extract of Pholiota adiposa, or a polyphenol extract of Grifola frondosa. The preparation method of the Agrocybe oleifera polysaccharide extract comprises the following steps: (1) Mixing Agrocybe oleifera with water, performing extraction, filtering, and collecting the filtrate; (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 60-70° C. to obtain a concentrated solution; (3) Add 3-5 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let stand at 20-25°C for 10-14 hours, then centrifuge and discard the supernatant, collect the precipitate and dry it at 50-60°C to constant weight to obtain a polysaccharide extract of Agrocybe oleifera; The preparation method of the Pholiota adiposa polyphenol extract comprises the following steps: (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation; (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution; (3) drying the concentrated solution obtained in step (2) at 50-60° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract; The preparation method of the Grifola frondosa polyphenol extract comprises the following steps: (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant; (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution; (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

8. A method for preparing a polysaccharide extract of Agrocybe tumefaciens, comprising the following steps: (1) Mixing Agrocybe oleifera with water, performing extraction, filtering, and collecting the filtrate; (2) The filtrate obtained in step (1) was concentrated by rotary evaporation at 60-70° C. to obtain a concentrated solution; (3) Add 3-5 times the volume of anhydrous ethanol to the concentrated solution obtained in step (2), mix well, and let it stand at 20-25°C for 10-14 hours. Then, centrifuge and discard the supernatant. Collect the precipitate and dry it at 50-60°C to constant weight to obtain a polysaccharide extract of Agrocybe oleifera.

9. A method for preparing a Pholiota adiposa polyphenol extract, comprising the following steps: (1) extracting by mixing Pholiota adzuki and ethanol aqueous solution, and then collecting the supernatant by centrifugation; (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution; (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Pholiota adiposa polyphenol extract.

10. A method for preparing a Grifola frondosa polyphenol extract, comprising the following steps: (1) Mixing the maitake mushroom and ethanol aqueous solution for extraction, and then centrifuging to collect the supernatant; (2) The supernatant obtained in step (1) was concentrated by rotary evaporation at 50-60° C. to obtain a concentrated solution; (3) The concentrated solution obtained in step (2) is dried at 50-60° C. to a constant weight to obtain a Grifola frondosa polyphenol extract.

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