Preparation method of ganoderma lucidum powder extract rich in high-activity triterpenes and ganoderma lucidum biscuits

Abstract

The application belongs to the technical field of food, and particularly relates to a preparation method of ganoderma lucidum powder extract rich in high-activity triterpenes and ganoderma biscuit. The preparation method of the ganoderma lucidum powder extract rich in high-activity triterpenes comprises the following steps: (1) drying ganoderma lucidum entities and crushing to obtain ganoderma lucidum coarse powder; (2) adding an ethanol aqueous solution to the ganoderma lucidum coarse powder, heating and refluxing to extract, combining the extract, concentrating the extract, recovering ethanol, obtaining an alcohol extract, and collecting an extraction residue; (3) fermentation: adding glucose to the extraction residue, sterilizing, adding sterile water to obtain fermentation raw materials, continuously adding a compound bacteria, sterilizing after the fermentation is completed, centrifuging, taking supernatant to obtain a fermentation liquor; and (4) mixing the alcohol extract and the fermentation liquor, vacuum drying to obtain the ganoderma lucidum powder extract rich in high-activity triterpenes. The application uses the process of first alcohol extraction and then fermentation by using specific bacteria, and the ganoderma lucidum powder extract prepared by the process has high contents of triterpenes, polysaccharides and flavones, and excellent storage stability.

Description

Technical Field

[0001] This invention belongs to the field of food technology, specifically relating to a method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes and Ganoderma lucidum biscuits. Background Technology

[0002] Reishi mushroom (Ganoderma lucidum) is a traditional medicinal fungus widely recognized for its various biological activities, including boosting immunity, anti-oxidation, anti-tumor, anti-inflammatory, blood sugar-lowering, and liver-protecting effects. Modern scientific research shows that reishi is rich in various active ingredients, among which triterpenoids (ganodermic acids) and crude polysaccharides are considered important components contributing to its diverse pharmacological effects. Adding reishi powder extract to everyday foods not only enhances their nutritional value but also meets consumers' demand for natural, healthy, and convenient foods.

[0003] Currently, common processing forms of Ganoderma lucidum include Ganoderma lucidum spore powder, Ganoderma lucidum tablets, Ganoderma lucidum capsules, and Ganoderma lucidum tea. However, traditional processing methods often fail to effectively preserve the active triterpenoid components in Ganoderma lucidum, resulting in low bioavailability and limiting its application in functional foods and pharmaceuticals. Existing technologies often employ ethanol reflux extraction, which, while improving the extraction rate of triterpenoids to some extent, still suffers from low polysaccharide extraction efficiency. Chinese patent CN109010385B discloses a method for preparing Ganoderma lucidum powder, which improves the extraction rate of crude polysaccharides, but the extraction rate of active triterpenoid components remains unsatisfactory.

[0004] Currently, there is a lack of a method on the market that can simultaneously and efficiently extract active triterpenes and polysaccharides. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes and Ganoderma lucidum biscuits.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] A method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes includes the following steps:

[0008] (1) Dry the Ganoderma lucidum body and then pulverize it to obtain Ganoderma lucidum coarse powder;

[0009] (2) Add an ethanol-water solution to the crude Ganoderma lucidum powder, heat and reflux to extract, combine the extracts, and recover the ethanol to obtain the alcohol extract, and collect the extraction residue;

[0010] (3) Fermentation: Add glucose to the extraction residue, sterilize, add sterile water to obtain fermentation raw material, continue to add compound bacteria, after fermentation is completed, sterilize, centrifuge, and take the supernatant to obtain fermentation broth.

[0011] (4) Mix the alcohol extract and fermentation broth, and dry them to obtain Ganoderma lucidum powder extract rich in highly active triterpenes.

[0012] Preferably, in step (1), the Ganoderma lucidum body is dried and then pulverized to a size of less than 20 mesh.

[0013] Preferably, the concentration of the ethanol aqueous solution in step (2) is 50-60 wt%.

[0014] Preferably, in step (2), the heating and reflux extraction is performed 2-4 times, with each extraction lasting 1-2 hours.

[0015] This invention first extracts Ganoderma lucidum powder with alcohol, then ferments it. This combined alcohol extraction and fermentation process not only increases the content of active substances but also improves the storage stability of the extract. Analysis shows that the active components in the product obtained through this combined alcohol extraction and fermentation process have a synergistic protective effect, contain fewer hygroscopic components, have strong antioxidant capacity, and exhibit better product stability.

[0016] Preferably, the compound bacteria in step (3) include *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*.

[0017] Preferably, in step (3), the ratio of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in the compound bacteria is 1:(1.3-1.5):(0.4-0.7).

[0018] Preferably, in step (3), the dosage of the compound bacteria is 10. 8 -10 9 CFU / mL fermentation feedstock.

[0019] Preferably, the fermentation conditions in step (3) are: aerobic fermentation at 25-35℃ for 30-40 hours.

[0020] The strains selected in this invention can simultaneously increase the content of triterpenes and polysaccharides in the extract, while also improving the stability of Ganoderma lucidum powder extract.

[0021] This invention provides the application of the method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes in the preparation of Ganoderma lucidum biscuits.

[0022] Preferably, the Ganoderma lucidum biscuit comprises the following components in parts by weight: 45-55 parts white sugar, 90-100 parts palm oil, 0.5-1 part salt, 3-4 parts sodium bicarbonate, 0.3-1 part ammonium bicarbonate, 25-30 parts caramel, 25-30 parts brown sugar syrup, 14-16 parts water, 0.10-0.15 parts Ganoderma lucidum powder extract, and 350-400 parts low-gluten flour.

[0023] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0024] 1. Compared with traditional extraction processes, this invention employs alcohol-fermentation extraction technology, which effectively improves the extraction efficiency of triterpenoids, polysaccharides, and flavonoids in Ganoderma lucidum. The obtained Ganoderma lucidum powder extract contains significantly higher levels of triterpenoids, polysaccharides, and flavonoids than existing technologies, thereby enhancing its antioxidant, immunomodulatory, and hepatoprotective biological activities and improving the functional value of the product.

[0025] 2. The Ganoderma lucidum powder extract prepared by this invention exhibits good storage stability, effectively preventing the degradation and loss of active ingredients during storage. Simultaneously, the extract demonstrates good processing adaptability, enabling stable application in baked goods such as biscuits, extending the product's shelf life and functional properties, and showing promising prospects for industrial application. Detailed Implementation

[0026] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] All raw materials used in the following embodiments of the present invention are commercially available products:

[0028] Mature Ganoderma lucidum, Shandong Guansheng Ganoderma Co., Ltd. *Lactobacillus plantarum*, strain number: HZB220201; *Acetobacter xylose*, strain number: HZB196686; *Aspergillus niger*, strain number: HZB128480; *Lactobacillus gasseri*, strain number: HZB138217; purchased from Zhengzhou Fangjue Biotechnology Co., Ltd. *Lactobacillus rhamnosus*, product number: HZB115392; *Lactobacillus delbrueckii*, product number: HZB139506; *Lactobacillus plantarum*, product number: HZB116349; *Lactobacillus casei*, product number: HZB116347; *Lactobacillus bulgaricus*, product number: HZB116558; purchased from Hangzhou Branch of Wuhan Gray Algae Biotechnology Co., Ltd.

[0029] The ingredients used to make the biscuits are all commercially available products.

[0030] Example 1

[0031] This embodiment provides a method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes, including the following steps:

[0032] (1) After drying mature Ganoderma lucidum, it is pulverized to a finer mesh than 100 to obtain coarse Ganoderma lucidum powder.

[0033] (2) Add 6 times the mass of 55wt% ethanol aqueous solution to the crude Ganoderma lucidum powder, heat and reflux to extract 3 times, each time for 1.5h, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue.

[0034] (3) Add 1% glucose (by weight of the extraction residue) to the extraction residue, sterilize, and add 4 times the weight of sterile water to obtain the fermentation raw material. Continue to add a compound microbial culture, which includes *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*. The dosage of the compound microbial culture is 5 × 10⁻⁶. 8 The fermentation feed consisted of CFU / mL of a compound microbial culture containing *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a viable ratio of 1:1.4:0.6. Fermentation was carried out at 30℃ with aerobic conditions for 38 hours at an air flow rate of 0.6 m³ / (m³*min). After fermentation, the culture was sterilized, centrifuged, and the supernatant was collected to obtain the fermentation broth.

[0035] (4) Mix the alcohol extract and fermentation broth, and vacuum dry to a water content of 1.97 wt% to obtain Ganoderma lucidum powder extract rich in highly active triterpenes.

[0036] This embodiment provides a Ganoderma lucidum biscuit prepared using the Ganoderma lucidum powder extract rich in highly active triterpenes obtained by the above preparation method, comprising the following components in parts by weight: 50 parts white sugar, 100 parts palm oil, 1 part salt, 3 parts sodium bicarbonate, 1 part ammonium bicarbonate, 30 parts caramel, 30 parts brown sugar syrup, 16 parts water, 0.13 parts Ganoderma lucidum powder extract, and 350 parts low-gluten flour.

[0037] Example 2

[0038] This embodiment provides a method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes, including the following steps:

[0039] (1) After drying mature Ganoderma lucidum, it is pulverized to a finer mesh than 100 to obtain coarse Ganoderma lucidum powder.

[0040] (2) Add 6 times the mass of 50wt% ethanol aqueous solution to the crude Ganoderma lucidum powder, heat and reflux to extract 4 times, 1 hour each time, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue.

[0041] (3) Add 1% glucose (by weight of the extraction residue) to the extraction residue, sterilize, and add 3 times the weight of sterile water to obtain the fermentation raw material. Continue to add a compound microbial culture, including *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*. The dosage of the compound microbial culture is 10... 8The fermentation feedstock consisted of CFU / mL of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a viable ratio of 1:1.3:0.4. Fermentation was carried out at 30℃ with aerobic conditions for 35 hours at an air flow rate of 0.6 m³ / (m³*min). After fermentation, the mixture was sterilized, centrifuged, and the supernatant was collected to obtain the fermentation broth.

[0042] (4) Mix the alcohol extract and fermentation broth, and vacuum dry to a water content of 1.97 wt% to obtain Ganoderma lucidum powder extract rich in highly active triterpenes.

[0043] This embodiment provides a Ganoderma lucidum biscuit prepared using the Ganoderma lucidum powder extract rich in highly active triterpenes obtained by the above preparation method, comprising the following components in parts by weight: 50 parts white sugar, 100 parts palm oil, 1 part salt, 3 parts sodium bicarbonate, 1 part ammonium bicarbonate, 30 parts caramel, 30 parts brown sugar syrup, 16 parts water, 0.13 parts Ganoderma lucidum powder extract, and 350 parts low-gluten flour.

[0044] Example 3

[0045] This embodiment provides a method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes, including the following steps:

[0046] (1) After drying mature Ganoderma lucidum, it is pulverized to a finer mesh than 100 to obtain coarse Ganoderma lucidum powder.

[0047] (2) Add 6 times the mass of 60wt% ethanol aqueous solution to the crude Ganoderma lucidum powder, heat and reflux twice, extract for 2 hours each time, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue.

[0048] (3) Add 1% glucose (by weight of the extraction residue) to the extraction residue, sterilize, and add 5 times the weight of sterile water to obtain the fermentation raw material. Continue to add a compound microbial culture, which includes *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*. The dosage of the compound microbial culture is 10... 9 The fermentation feedstock consisted of CFU / mL of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a viable ratio of 1:1.5:0.7. Fermentation was carried out at 30℃ with aerobic conditions for 40 hours at an air flow rate of 0.6 m³ / (m³*min). After fermentation, the mixture was sterilized, centrifuged, and the supernatant was collected to obtain the fermentation broth.

[0049] (4) Mix the alcohol extract and fermentation broth, and vacuum dry to a water content of 1.97 wt% to obtain Ganoderma lucidum powder extract rich in highly active triterpenes.

[0050] This embodiment provides a Ganoderma lucidum biscuit prepared using the Ganoderma lucidum powder extract rich in highly active triterpenes obtained by the above preparation method, comprising the following components in parts by weight: 50 parts white sugar, 100 parts palm oil, 1 part salt, 3 parts sodium bicarbonate, 1 part ammonium bicarbonate, 30 parts caramel, 30 parts brown sugar syrup, 16 parts water, 0.13 parts Ganoderma lucidum powder extract, and 350 parts low-gluten flour.

[0051] Comparative Example 1

[0052] The difference between this comparative example and Example 1 is that fermentation is performed first, followed by alcohol extraction.

[0053] A method for preparing Ganoderma lucidum powder extract rich in highly active triterpenes includes the following steps:

[0054] (1) After drying mature Ganoderma lucidum, it is pulverized to a finer mesh than 100 to obtain coarse Ganoderma lucidum powder;

[0055] (2) Add 1% glucose (by weight of the crude Ganoderma lucidum powder) to the crude Ganoderma lucidum powder, sterilize, and add sterile water at 4 times the weight of the crude Ganoderma lucidum powder to obtain the fermentation raw material. Continue to add compound bacteria, which include *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*. The dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was prepared at CFU / mL. The ratio of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in the compound bacteria was 1:1.4:0.6. The fermentation was carried out at 30℃ for 38 hours with an air flow rate of 0.6 cubic meters per minute. After fermentation, the mixture was sterilized, centrifuged, and the supernatant was collected to obtain the fermentation broth. The fermentation residue was also collected.

[0056] (3) Add 6 times the mass of 55wt% ethanol to the fermentation residue, heat and reflux to extract 3 times, each time for 1.5h, combine the extracts, recover the ethanol, and obtain the alcohol extract.

[0057] (4) Mix the alcohol extract and fermentation broth, and vacuum dry to a water content of 1.97 wt% to obtain Ganoderma lucidum powder extract rich in highly active triterpenes.

[0058] Comparative Example 2

[0059] The difference between this comparative example and Example 1 is that the compound bacteria include Lactobacillus bulgaricus, Lactobacillus casei, and Lactobacillus delbrueckii, and the dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was prepared at CFU / mL, with the ratio of viable Lactobacillus bulgaricus, Lactobacillus casei, and Lactobacillus delbrueckii in the compound bacteria being 1:1.3:0.4; and anaerobic fermentation was carried out at 37℃ for 38 hours.

[0060] Comparative Example 3

[0061] The difference between this comparative example and Example 1 is that the compound bacteria include Lactobacillus plantarum, Lactobacillus gasseri, and Lactobacillus rhamnosus, and the dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was CFU / mL, and the ratio of viable Lactobacillus plantarum, Lactobacillus gasseri, and Lactobacillus rhamnosus in the compound bacteria was 1:1.3:0.4; anaerobic fermentation was carried out at 37℃ for 38 hours.

[0062] Comparative Example 4

[0063] The difference between this comparative example and Example 1 is that the compound bacteria were replaced with Aspergillus niger, and the amount of Aspergillus niger was 5 × 10⁻⁶. 8 CFU / mL fermentation feedstock; aerobic fermentation at 30℃ for 38 hours.

[0064] Comparative Example 5

[0065] The difference between this comparative example and Example 1 is that the compound bacteria include *Lactobacillus plantarum* and *Aspergillus niger*, and the dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was CFU / mL, and the ratio of viable Lactobacillus plantarum to Aspergillus niger in the compound bacteria was 1.4:0.6; aerobic fermentation was carried out at 30℃ for 38 hours.

[0066] Comparative Example 6

[0067] The difference between this comparative example and Example 1 is that the compound bacteria include *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*, and the dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was prepared at CFU / mL, with the ratio of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in the complex culture being 0.6:1:1.4; fermentation was carried out at 30℃ for 38 hours under aerobic conditions.

[0068] Comparative Example 7

[0069] The compound bacteria include *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger*, and the dosage of the compound bacteria is 5 × 10⁻⁶. 8 The fermentation feedstock was prepared at CFU / mL, with the ratio of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in the complex culture being 1.4:0.6:1; fermentation was carried out at 30℃ for 38 hours under aerobic conditions.

[0070] Performance testing

[0071] The contents of triterpenoids, polysaccharides and flavonoids in the Ganoderma lucidum powder extracts prepared in Examples 1-3 and Comparative Examples 1-7 were determined according to the method in "Changes in the Main Active Substances of Fermented Ganoderma lucidum and its Inhibitory Effect on HepG2 Cells".

[0072] The Ganoderma lucidum powder extracts prepared in Examples 1-3 and Comparative Examples 1-7 were stored at 40℃±2℃ / 75%RH±5%RH in a light-proof and sealed manner for 3 months, and it was observed whether clumping occurred.

[0073] The results are shown in Table 1-2.

[0074] Table 1 Test Results

[0075]

[0076]

[0077] Table 2 Storage Stability

[0078] Example 2 Uniform and free of lumps Example 3 Uniform and free of lumps Comparative Example 1 Large clumps, difficult to disperse Comparative Example 2 Light clumps easily disperse Comparative Example 3 Light clumps easily disperse Comparative Example 4 Large clumps, difficult to disperse Comparative Example 5 Light clumps easily disperse Comparative Example 6 Uniform and free of lumps Comparative Example 7 Uniform and free of lumps

[0079] As shown in Table 1, the Ganoderma lucidum powders in Examples 1-3 have high contents of triterpenes, polysaccharides and flavonoids, and good storage stability.

[0080] In Comparative Example 1, the process of fermentation followed by alcohol extraction resulted in a decrease in the content of triterpenes, polysaccharides, and flavonoids, as well as a decrease in storage stability. This indicates that the order of alcohol extraction and fermentation has a significant impact on the composition of the active components of the product.

[0081] Comparative Examples 2 and 3 used common probiotics for fermentation, and the results were worse than those of the strains used in this invention, especially the polysaccharide content was significantly reduced, indicating that the fermentation strain has an important influence on the polysaccharide content.

[0082] Comparative Examples 4-5 illustrate that the three strains selected in this invention, when combined and fermented, achieve better results.

[0083] In Comparative Examples 6-7, fermentation with different proportions of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* resulted in a decrease in the contents of triterpenes, polysaccharides, and flavonoids, especially a significant decrease in polysaccharides. This indicates that the type and proportion of microbial strains have a significant impact on the polysaccharide content in *Ganoderma lucidum* powder extract. Analysis shows that fermentation with the microbial strains formulated according to this invention is more effective in disrupting the mycelial structure and surface structure of *Ganoderma lucidum* after alcohol extraction, resulting in the release of more active substances, while simultaneously protecting the active polysaccharide chains from damage.

[0084] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The application of the combination of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in enhancing the triterpenoid content in *Ganoderma lucidum* powder extract, characterized by: The preparation methods for Ganoderma lucidum powder extract include: (1) Dry the Ganoderma lucidum body and then pulverize it to obtain Ganoderma lucidum coarse powder; (2) Add an aqueous ethanol solution to the crude Ganoderma lucidum powder, heat and reflux to extract, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue; (3) Add glucose and water to the extraction residue to obtain fermentation raw material, continue to add compound bacteria, carry out aerobic fermentation, sterilize after fermentation, centrifuge, take the supernatant to obtain fermentation liquid; (4) Mix the alcohol extract and fermentation broth, dry them, and obtain Ganoderma lucidum powder extract; The compound bacteria consist of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a ratio of 1:(1.3-1.5):(0.4-0.7). Lactobacillus plantarum, strain number: HZB220201; Acetobacter xylose, strain number: HZB196686; Aspergillus niger, strain number: HZB128480; all were obtained from Zhengzhou Fangjue Biotechnology Co., Ltd.

2. The application of the combination of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in increasing the polysaccharide content in *Ganoderma lucidum* powder extract, characterized by: The preparation methods for Ganoderma lucidum powder extract include: (1) Dry the Ganoderma lucidum body and then pulverize it to obtain Ganoderma lucidum coarse powder; (2) Add an aqueous ethanol solution to the crude Ganoderma lucidum powder, heat and reflux to extract, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue; (3) Add glucose and water to the extraction residue to obtain fermentation raw material, continue to add compound bacteria, carry out aerobic fermentation, sterilize after fermentation, centrifuge, take the supernatant to obtain fermentation liquid; (4) Mix the alcohol extract and fermentation broth, dry them, and obtain Ganoderma lucidum powder extract; The compound bacteria consist of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a ratio of 1:(1.3-1.5):(0.4-0.7). Lactobacillus plantarum, strain number: HZB220201; Acetobacter xylose, strain number: HZB196686; Aspergillus niger, strain number: HZB128480; all were obtained from Zhengzhou Fangjue Biotechnology Co., Ltd.

3. The application of the combination of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in enhancing the flavonoid content in *Ganoderma lucidum* powder extract, characterized by: The preparation methods for Ganoderma lucidum powder extract include: (1) Dry the Ganoderma lucidum body and then pulverize it to obtain Ganoderma lucidum coarse powder; (2) Add an aqueous ethanol solution to the crude Ganoderma lucidum powder, heat and reflux to extract, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue; (3) Add glucose and water to the extraction residue to obtain fermentation raw material, continue to add compound bacteria, carry out aerobic fermentation, sterilize after fermentation, centrifuge, take the supernatant to obtain fermentation liquid; (4) Mix the alcohol extract and fermentation broth, dry them, and obtain Ganoderma lucidum powder extract; The compound bacteria consist of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a ratio of 1:(1.3-1.5):(0.4-0.7). Lactobacillus plantarum, strain number: HZB220201; Acetobacter xylose, strain number: HZB196686; Aspergillus niger, strain number: HZB128480; all were obtained from Zhengzhou Fangjue Biotechnology Co., Ltd.

4. The application of the combination of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in enhancing the prevention of clumping in Ganoderma lucidum powder extract, characterized by: The preparation methods for Ganoderma lucidum powder extract include: (1) Dry the Ganoderma lucidum body and then pulverize it to obtain Ganoderma lucidum coarse powder; (2) Add an aqueous ethanol solution to the crude Ganoderma lucidum powder, heat and reflux to extract, combine the extracts, recover the ethanol to obtain the alcohol extract, and collect the extraction residue; (3) Add glucose and water to the extraction residue to obtain fermentation raw material, continue to add compound bacteria, carry out aerobic fermentation, sterilize after fermentation, centrifuge, take the supernatant to obtain fermentation liquid; (4) Mix the alcohol extract and fermentation broth, dry them, and obtain Ganoderma lucidum powder extract; The compound bacteria consist of viable bacteria of *Acetobacter xylose*, *Lactobacillus plantarum*, and *Aspergillus niger* in a ratio of 1:(1.3-1.5):(0.4-0.7). Lactobacillus plantarum, strain number: HZB220201; Acetobacter xylose, strain number: HZB196686; Aspergillus niger, strain number: HZB128480; all were obtained from Zhengzhou Fangjue Biotechnology Co., Ltd.

5. The application according to any one of claims 1-4, characterized in that: In step (1), the Ganoderma lucidum body is dried and then pulverized to a size of less than 20 mesh.

6. The application according to any one of claims 1-4, characterized in that: The concentration of the ethanol aqueous solution in step (2) is 50-60 wt%.

7. The application according to any one of claims 1-4, characterized in that: In step (2), the extraction is performed by heating and reflux 2-4 times, with each extraction lasting 1-2 hours.

8. The application according to any one of claims 1-4, characterized in that: In step (3), the dosage of the compound bacteria is 10. 8 -10 9 CFU / mL fermentation feedstock.

9. The application according to any one of claims 1-4, characterized in that: The fermentation conditions in step (3) are: aerobic fermentation at 25-35℃ for 30-40 hours.

10. The application according to any one of claims 1-4, characterized in that: The Ganoderma lucidum powder extract is used to prepare Ganoderma lucidum biscuits.

11. The application according to claim 10, characterized in that, The Ganoderma lucidum biscuits comprise the following components in parts by weight: 45-55 parts white sugar, 90-100 parts palm oil, 0.5-1 part salt, 3-4 parts sodium bicarbonate, 0.3-1 part ammonium bicarbonate, 25-30 parts caramel, 25-30 parts brown sugar syrup, 14-16 parts water, 0.10-0.15 parts Ganoderma lucidum powder extract, and 350-400 parts low-gluten flour.

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