Method for improving quality of ganoderma lucidum mushroom residues through synergistic fermentation of bacteria and enzymes and application of method

Abstract

The invention discloses a method for improving the quality of ganoderma lucidum mushroom residues through synergistic fermentation of bacteria and enzymes and application of the method. According to the method disclosed by the invention, cellulase, beta-galactosidase and xylanase with proper concentrations and lactobacillus plantarum, bacillus subtilis and saccharomyces cerevisiae with proper proportions are adopted, and enzymes and bacteria are mutually cooperated, so that the characteristics of the raw materials are well balanced, the finally prepared ganoderma lucidum mushroom bran is high in protein content and low in fiber content, and the nutritional value of a feed is improved; the total amount of amino acids and essential amino acids are remarkably increased, and the feed protein quality can be improved; the rumen dry matter digestibility and energy metabolites are remarkably improved, and ruminant absorption and utilization are facilitated; the ganoderma lucidum mushroom bran fermented feed is added into the daily ration, so that the daily gain, the carcass rate and the dressing percentage of fed animals can be remarkably improved, the levels of serum GLU, TG, ALT and AST are reduced, the expression of key inflammatory cytokines (IL-1beta and IL-6) is inhibited, and the ganoderma lucidum mushroom bran fermented feed has the comprehensive effects of regulating glycolipid metabolism and relieving liver load and systemic inflammation, so that the immune function of animal bodies is improved.

Description

Technical Field

[0001] This invention belongs to the field of microbial preparation technology, specifically relating to a method for improving the quality of Ganoderma lucidum fungus residue through synergistic fermentation of bacteria and enzymes, and its application. Background Technology

[0002] Feed is the material foundation for the development of animal husbandry. With the increasing number of livestock and poultry and the annual output of livestock and poultry products, as well as the rapid population growth, the contradiction between humans and livestock competing for food has become increasingly prominent. Feed shortages have become a key bottleneck restricting the sustainable development of my country's animal husbandry. my country has a vast territory and abundant resources, possessing a wide variety of non-grain feed resources. However, these non-grain feed resources have complex compositions, unbalanced nutrition, and often contain various anti-nutritional factors and mycotoxins. They cannot be used directly without treatment, and even with simple processing, their use must be strictly limited. Furthermore, they have poor palatability and low feed value, and their nutritional composition varies greatly due to the combined effects of factors such as origin, processing, and storage conditions, making it difficult to maintain stable quality. Therefore, they have not been fully and effectively utilized in animal husbandry production.

[0003] Ganoderma lucidum mycelial residue refers to the culture left after cultivating Ganoderma lucidum using materials such as mushroom straw, wheat bran, corn cob, sugarcane bagasse, peanut vines, and sweet potato vines. It is the mycelial residue and metabolic products of Ganoderma lucidum and is an important non-grain feed resource. In addition to being rich in nutrients such as crude protein, amino acids, and sugars, Ganoderma lucidum mycelial residue also contains a large amount of cellulose, hemicellulose, lignin, and other substances that are difficult for livestock and poultry to digest and absorb. Simply enzymatically hydrolyzing Ganoderma lucidum mycelial residue results in incomplete degradation of lignocellulose, a lack of functional active substances, and a lack of subsequent biotransformation, leading to low release of reducing sugars. The oligosaccharides and monosaccharides produced cannot be further utilized and instead provide a breeding ground for miscellaneous bacteria. The product lacks acidity and aroma, has poor palatability, and is deficient in organic acids, small peptides, vitamins, and extracellular polysaccharides produced by microbial metabolism, resulting in significantly low antioxidant and antibacterial biological activities. Furthermore, the fermentation of Ganoderma lucidum fungus residue using only compound microorganisms is hampered by the cell wall barrier: lignocellulose is difficult for microbial enzymes to effectively break down, limiting the improvement in cellulose degradation rate; fermentable sugars are released slowly, carbon sources are trapped in the cell wall, probiotic proliferation is delayed, pH decrease is postponed, and the risk of contamination by other microorganisms increases accordingly; phytic acid and other anti-nutritional factors remain in high levels, animal utilization remains low, and the increase in feed value is limited, resulting in an overall predicament of "incomplete fermentation and difficulty in improving quality." Therefore, it is necessary to develop a method for improving the quality of Ganoderma lucidum fungus residue through the synergistic fermentation of compound enzymes and compound microorganisms, in order to improve fermentation efficiency, inhibit the growth of other microorganisms, improve feed quality, and facilitate the application of Ganoderma lucidum fungus residue in feed. Summary of the Invention

[0004] Based on this, the present invention provides a method for improving the quality of Ganoderma lucidum mycelium residue through synergistic fermentation of bacteria and enzymes, thus solving the problems mentioned in the background art.

[0005] A method for improving the quality of Ganoderma lucidum mycelium residue through synergistic fermentation of microorganisms and enzymes includes the following steps: S1. Take clean, mold-free Ganoderma lucidum spore sticks after harvesting, crush them, air dry them naturally, pulverize them, and sieve them to obtain Ganoderma lucidum spore powder.

[0006] S2. Mix 1000~3000 U / g of cellulase, 80~160 U / g of β-galactosidase and 20000~40000 U / g of xylanase evenly, add water to dissolve completely, and activate at 40~55℃ for 35~45 min.

[0007] S3. Add the Ganoderma lucidum fungus powder obtained in step S1 to the compound enzyme preparation activated in step S2, with a material-to-liquid ratio of (0.8~1.4):(0.6~1.2), and mix thoroughly to obtain a mixture of Ganoderma lucidum fungus powder and compound enzyme preparation.

[0008] S4. Add 15-25 g / kg of carbon source and activated Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae with mass fractions of 8%-12%, 8%-12% and 2%-12% respectively to the mixture of Ganoderma lucidum fungus bran and compound enzyme preparation obtained in step S3. After stirring evenly, anaerobic ferment at 30-37℃ for 40-96 hours.

[0009] S5. Dry, crush, and sieve the Ganoderma lucidum mycelium residue obtained from fermentation in step S4 to obtain Ganoderma lucidum mycelium residue feed with synergistic fermentation of mycelium and enzyme.

[0010] Preferably, the sieving in step S1 is sieving through a 40-80 mesh sieve.

[0011] Preferably, in step S2, the contents of cellulase, β-galactosidase, and xylanase are 3000 U / g, 160 U / g, and 40000 U / g, respectively.

[0012] Preferably, in step S4, the mass fractions of Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae are 8%, 8%, and 12%, respectively.

[0013] Preferably, the fermentation temperature in step S4 is 35°C and the fermentation time is 96 hours.

[0014] Preferably, the carbon source in step S4 is corn flour.

[0015] The Ganoderma lucidum mycelium bran prepared by any of the above preparation methods.

[0016] The above-mentioned application of Ganoderma lucidum fungus residue in the preparation of ruminant animal feed.

[0017] The present invention has the following beneficial effects: The method of this invention cleverly combines the advantages of enzymatic hydrolysis and microbial fermentation. It overcomes the problems of "incomplete degradation of lignocellulose, lack of functional active substances and lack of subsequent biotransformation, resulting in low release of reducing sugars" in simple enzymatic hydrolysis, and solves the technical problems of "lignocellulose is difficult to be effectively broken down by the microbial enzyme system itself, resulting in limited improvement in cellulose degradation rate; slow release of fermentable sugars and increased risk of contamination by miscellaneous bacteria" in simple compound microbial fermentation.

[0018] The combination and dosage of enzyme preparations, the inoculation amount and ratio of synergistic fermentation strains, and the fermentation process of this invention all affect the nutritional composition of Ganoderma lucidum fungus bran fermented feed: First, Ganoderma lucidum fungus bran itself contains residual functional active substances or immune enhancers such as Ganoderma lucidum polysaccharides, triterpenes, and sterols, and the polysaccharide content can still remain high after fermentation; Second, the natural antibacterial factors in Ganoderma lucidum fungus bran synergistically reduce the toxins in the finished fermented feed with the acid production of Lactobacillus; At the same time, a large number of pharmacologically active ingredients such as Ganoderma lucidum polysaccharides remain after fermentation of Ganoderma lucidum fungus bran, which can enhance animal immune function and improve animal production performance.

[0019] The various process steps in this invention are inseparable and synergistic, forming an indispensable whole. Only under the raw material ratios and process conditions of this invention, namely, appropriate concentrations of cellulase, β-galactosidase, and xylanase, and appropriate ratios of Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae, can the enzymes and bacteria work synergistically to effectively balance the characteristics of each raw material. This results in a final product of Ganoderma lucidum bran with high protein content and low fiber content, improving the nutritional value of the feed. The total amino acid content and essential amino acids are significantly increased, which helps improve the quality of feed protein. It also significantly improves rumen dry matter digestibility and energy metabolites (VFA), which are beneficial for ruminants to absorb and utilize, making it a potential feed for ruminants. Adding fermented Ganoderma lucidum bran to the diet can significantly improve the daily weight gain, carcass rate, and slaughter rate of the animals, improving overall production performance. At the same time, it downregulates serum GLU, TG, ALT, and AST levels and inhibits the expression of key inflammatory cytokines (IL-1β, IL-6), exhibiting a comprehensive effect of regulating glucose and lipid metabolism, reducing liver burden and systemic inflammation, thereby improving the animal's immune function.

[0020] The core function of corn flour in this invention is to act as a "quick-acting carbon source," which is preferentially utilized by microorganisms to rapidly increase the carbon / nitrogen (C / N) ratio of the culture medium, thereby significantly promoting mycelial growth and enzyme secretion. The specific benefits are as follows: 1. Rapidly provides fermentable sugars: Corn flour is rich in starch, which, after being hydrolyzed by added or secreted amylase from Ganoderma lucidum itself, can be continuously converted into reducing sugars such as glucose and maltose, providing fast-acting energy for Ganoderma lucidum mycelium and synergistic strains (such as yeast and lactic acid bacteria), shortening the fermentation lag period, and increasing biomass; 2. Optimizes the C / N ratio: After supplementing with corn flour, the C / N ratio can be increased from about 15:1 to 25~30:1, which is closer to the optimal range for high production of polysaccharides, triterpenes, and other secondary metabolites in Ganoderma lucidum, thus significantly increasing the content of polysaccharides, soluble proteins, and total amino acids in Ganoderma lucidum; 3. Balances economy and palatability: Corn flour is inexpensive and widely available; adding 5%~15% can significantly shorten the fermentation cycle by 20%~30%, and give the fermented material a pleasant wine aroma, improving its palatability and commercial value as a functional feed or fertilizer. Detailed Implementation

[0021] To better understand the present invention, the following detailed description is provided in conjunction with embodiments. However, those skilled in the art will understand that the following embodiments are not intended to limit the scope of protection of the present invention, and any changes and modifications made based on the present invention are within the scope of protection of the present invention.

[0022] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0023] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0024] Experimental materials: The Ganoderma lucidum fungus residue used in this invention was provided by the National Engineering Research Center for Juncao (a type of fungus).

[0025] Cellulase (100,000 U / g), β-galactosidase (50,000 U / g), and xylanase (100,000 U / g) used in the experiment were purchased from Nanning Dongheng Huadao Biotechnology Co., Ltd.

[0026] The test strains Bacillus subtilis CGMCC 1.1086 and Lactobacillus plantarum CGMCC 1.557 were purchased from the Guangdong Provincial Microbial Culture Collection Center.

[0027] The brewing yeast used in this invention is Angel Yeast Co., Ltd.'s Fubang Private High-Activity Dry Yeast (for Livestock and Poultry).

[0028] The experimental method is as follows: Step 1: Take clean, mold-free Ganoderma lucidum spawn after harvesting Ganoderma lucidum once, mechanically crush it, air dry it naturally, pulverize it through an 80-mesh sieve, and store it for later use.

[0029] Step 2: Mix cellulase, β-galactosidase and xylanase in different proportions, add water to dissolve the compound enzyme preparation, stir thoroughly and mix well, and activate at 50℃ for 45 minutes.

[0030] Step 3: Add the Ganoderma lucidum mycelium powder from Step 1 to the activated compound enzyme solution at a material-to-liquid ratio of 1.2:1. After thorough mixing, pack the mixture into an anaerobic fermentation bag and incubate at 50°C for 24 hours.

[0031] Step 4: Determine the content of acid detergent fiber (ADF) and neutral detergent fiber (NDF) in the Ganoderma lucidum mycelium residue after enzymatic hydrolysis.

[0032] Step 5: Process the enzymatically hydrolyzed Ganoderma lucidum mycelium residue from Step 3 and determine its polysaccharide and reducing sugar content.

[0033] Table 1 L9 (3) 4 Orthogonal experimental design table for enzymatic hydrolysis of Ganoderma lucidum mycelium residue using compound enzymes

[0034] By designing an orthogonal experimental design for complex enzymes (see Table 1 for details), the optimal combination for enzymatic hydrolysis of Ganoderma lucidum mycelium residue was selected.

[0035] Using the degradation rate of acidic detergent fiber (A), the degradation rate of neutral detergent fiber (N), and the content of reducing sugar (S) in the Ganoderma lucidum mycelium residue after enzymatic hydrolysis as the main indicators, the overall enzymatic hydrolysis effect E of the mycelium residue was calculated by weighted method (E=0.35×A+0.35×N+0.30×S) (see Table 2 for details).

[0036] Table 2 L9 (3) 4 Results of orthogonal experiment on enzymatic hydrolysis of Ganoderma lucidum fungus residue with compound enzyme

[0037] Table 3. Analysis of the results of the orthogonal experiment on the enzymatic hydrolysis of Ganoderma lucidum mycelium residue by compound enzymes.

[0038] Using the effect of compound enzymatic hydrolysis of Ganoderma lucidum mycelium residue as the response value, the optimal enzymatic hydrolysis combination was obtained based on the orthogonal experimental results (see Table 3 for details): cellulase 3,000 U / g, β-galactosidase 160 U / g, xylanase 40,000 U / g; under the conditions of enzymatic hydrolysis time of 24 h, material-to-liquid ratio of 1.2:1, and temperature of 50℃, the highest degradation rate of acid detergent fiber was 11.30%, the highest degradation rate of neutral detergent fiber was 17.08%, and the highest reducing sugar content was 209.31 mg / g.

[0039] Based on the above results, the optimal enzymatic combination and conditions for the combined enzymatic hydrolysis of Ganoderma lucidum mycelium residue were determined to be the best response value for subsequent co-fermentation experiments on Ganoderma lucidum mycelium residue. The conditions for combined enzymatic hydrolysis of Ganoderma lucidum mycelium residue were: hydrolysis time of 24 h, material-to-liquid ratio of 1.2:1, cellulase 3,000 U / g, β-galactosidase 160 U / g, and xylanase 40,000 U / g.

[0040] In this experiment, Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae were used as compound microbial fermentation agents for solid-state fermentation of Ganoderma lucidum fungus bran. An orthogonal experimental design with three factors and three levels was adopted. The three factors were Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae, and the three levels were different inoculum amounts of the three bacteria: 2%, 8%, and 12%. The experimental design is shown in Table 4. Each experiment was conducted in triplicate.

[0041] Table 4 Orthogonal Table of Compound Microbial Combinations

[0042] Fermenting Ganoderma lucidum fungus residue using compound microbial agents includes the following steps.

[0043] Step 1: Take clean, mold-free Ganoderma lucidum spawn after harvesting Ganoderma lucidum once, mechanically crush it, air dry it naturally, pulverize it through an 80-mesh sieve, and store it for later use.

[0044] Step 2: Add 20g of corn flour to each kilogram of Ganoderma lucidum mycelium residue and mix thoroughly. Then add activated Lactobacillus plantarum (live count > 10). 9 CFU / mL) 20 mL~60 mL, activated Bacillus subtilis (>10) 9 CFU / mL) 60 mL~180 mL, activated brewer's yeast (>10) 9 Mix 25 mL to 75 mL of (CFU / mL) thoroughly, place in an anaerobic fermentation bag, and carry out anaerobic fermentation at 35℃ for 4 days.

[0045] Step 3: Collect and dry the fermented residue obtained in Step 2, and test its crude protein, neutral detergent fiber and acid detergent fiber content, and then calculate the acid detergent degradation rate and neutral detergent fiber degradation rate.

[0046] Through orthogonal experimental design of compound microorganisms, a better compound microbial combination for fermenting Ganoderma lucidum spores was selected (see Table 5 for details).

[0047] The optimal combination was selected based on the increase in crude protein content (P), acid detergent fiber degradation rate (A), and neutral detergent fiber degradation rate (N) in Ganoderma lucidum fungus bran after compound microbial fermentation (see Table 6 for details).

[0048] Table 5. Results of the orthogonal experiment on the fermentation of Ganoderma lucidum residue by compound microorganisms

[0049] Table 6. Analysis of the results of the orthogonal experiment on the fermentation of Ganoderma lucidum fungus residue by compound microorganisms.

[0050] Based on the orthogonal experiment results of compound enzyme fermentation of Ganoderma lucidum spores, three optimal microbial combinations were obtained: 8% inoculum of Lactobacillus plantarum, 12% inoculum of Bacillus subtilis, and 12% inoculum of Saccharomyces cerevisiae; 12% inoculum of Lactobacillus plantarum, 12% inoculum of Bacillus subtilis, and 12% inoculum of Saccharomyces cerevisiae; and 2% inoculum of Lactobacillus plantarum, 8% inoculum of Bacillus subtilis, and 2% inoculum of Saccharomyces cerevisiae; solid-state fermentation at 35℃ for 4 days.

[0051] Example 1 uses a compound enzyme to enzymatically hydrolyze Ganoderma lucidum mycelium residue, and obtains the best combination effect of enzymatic hydrolysis of Ganoderma lucidum mycelium residue; Example 2 is the screening of the optimal combination of compound microbial fermentation of Ganoderma lucidum mycelium residue.

[0052] This experiment utilizes the enzymatic hydrolysis combination in Example 1 and the compound microbial combination in Example 2 to conduct a synergistic fermentation experiment of Ganoderma lucidum mycelium residue by microorganisms and enzymes, including the following steps.

[0053] Step 1: Take clean, mold-free Ganoderma lucidum spawn after harvesting Ganoderma lucidum once, mechanically crush it, air dry it naturally, pulverize it through an 80-mesh sieve, and store it for later use.

[0054] Step 2: Mix cellulase, β-galactosidase and xylanase at concentrations of 3,000 U / g, 160 U / g and 40,000 U / g respectively, add water to dissolve them completely, and then activate them at 50°C for 45 min.

[0055] Step 3: Add the Ganoderma lucidum fungus powder from Step 1 to the activated compound enzyme preparation at a ratio of 1.2:1, mix thoroughly to obtain a mixture of Ganoderma lucidum fungus powder and compound enzyme preparation.

[0056] Step 4: Add 20g of corn flour, activated Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae to each kilogram of Ganoderma lucidum fungus and compound enzyme preparation obtained in Step 3. The inoculation amount is added according to the three ratios in Example 2 (Table 7). After thorough mixing, put it into a fermentation bag and carry out anaerobic fermentation at 35℃ for 4 days.

[0057] Step 5: Dry, crush, and sieve the Ganoderma lucidum mycelium residue obtained from step 4 fermentation to obtain Ganoderma lucidum mycelium residue feed with synergistic fermentation of mycelium and enzyme.

[0058] Table 7. Inoculum Size of Compound Microorganisms

[0059] Table 8. Co-fermentation of Ganoderma lucidum mycelium residue by microorganisms and enzymes

[0060] Nutritional composition comparison between raw Ganoderma lucidum mycelium residue (control group) that was neither enzymatically hydrolyzed nor fermented by microorganisms and using a microbial-enzyme co-fermentation method: The optimal enzymatic hydrolysis combination in Example 1, along with three levels of microbial fermentation with good results (see Table 7), was used to conduct synergistic fermentation of Ganoderma lucidum bran feed. The nutritional components, amino acid content, crude polysaccharide content, and in vitro fermentation performance were determined.

[0061] According to the experimental results (see Tables 8, 9, 10, and 11): synergistic fermentation of bacteria and enzymes significantly increases protein and reduces fiber, thus improving the nutritional value of feed; the total amount of amino acids and essential amino acids are significantly increased, which helps to improve the protein quality of feed; synergistic fermentation of bacteria and enzymes significantly improves rumen dry matter digestibility and energy metabolites (VFA), which is beneficial for ruminants to absorb and utilize, and has the potential to be used as ruminant feed.

[0062] Table 9. Analysis of conventional nutritional components of fermented Ganoderma lucidum mycelium residue under different treatments.

[0063] Note: Different letters in the same row indicate significant differences (p<0.05). Table 10. Amino acid and crude polysaccharide content of fermented Ganoderma lucidum mycelium residue under different treatments

[0064] Table 11. In vitro fermentation performance analysis of Ganoderma lucidum mycelium residue under different treatments

[0065] Example 4: Effect of feeding animals with Ganoderma lucidum mycelium residue fermented with enzymes and bacteria. Three-month-old Tibetan sheep were fed with Ganoderma lucidum mycelium bran feed obtained by synergistic fermentation of Ganoderma lucidum mycelium bran feed with inoculation amounts of 8% Lactobacillus plantarum, 12% Bacillus subtilis, and 12% Saccharomyces cerevisiae, as described in Example 3. The control group did not receive any Ganoderma lucidum mycelium bran fermented feed. The experimental group received 100g / head / day of the feed, and the experimental group received 200g / head / day of the feed. The experiment lasted for 60 days.

[0066] The experimental results (Tables 12, 13, and 14) show that adding Ganoderma lucidum bran fermented feed to the diet can significantly improve the daily weight gain, carcass rate, and slaughter rate of sheep, and improve overall production performance. At the same time, it downregulates serum GLU, TG, ALT, and AST levels and inhibits the expression of key inflammatory cytokines (IL-1β and IL-6), suggesting that it may have a comprehensive effect of regulating glucose and lipid metabolism, reducing liver burden and systemic inflammation, thereby improving the immune function of animals.

[0067] Table 12. Changes in body weight and production performance of experimental sheep during the experiment.

[0068] Note: Different letters in the same row indicate significant differences (p<0.05). Table 13 Physiological and biochemical indicators of experimental sheep serum

[0069] Note: Different letters in the same row indicate significant differences (p<0.05). Table 14 Cytokines in the serum of the experimental sheep

[0070] Note: Different letters in the same row indicate significant differences (p<0.05). While specific embodiments of the present invention have been described above, those skilled in the art should understand that the specific embodiments described are merely illustrative and not intended to limit the scope of the present invention. Equivalent modifications and variations made by those skilled in the art in accordance with the spirit of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A method for improving the quality of Ganoderma lucidum mycelium residue through synergistic fermentation of bacteria and enzymes, and its application, characterized in that, Includes the following steps: S1. Take clean, mold-free Ganoderma lucidum spawn after harvesting Ganoderma lucidum, crush it, air dry it naturally, pulverize it and sieve it to obtain Ganoderma lucidum spawn powder; S2. Mix 1000~3000 U / g of cellulase, 80~160 U / g of β-galactosidase and 20000~40000 U / g of xylanase evenly, add water to dissolve completely, and activate at 40~55℃ for 35~45 min. S3. Add the Ganoderma lucidum fungus powder obtained in step S1 to the compound enzyme preparation activated in step S2, with a material-to-liquid ratio of (0.8~1.4):(0.6~1.2), mix thoroughly to obtain a mixture of Ganoderma lucidum fungus powder and compound enzyme preparation; S4. Add 15-25 g / kg of carbon source and activated Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae with mass fractions of 8%-12%, 8%-12% and 2%-12% respectively to the mixture of Ganoderma lucidum fungus bran and compound enzyme preparation obtained in step S3. After stirring evenly, anaerobic ferment at 30-37℃ for 40-96 hours. S5. Dry, crush, and sieve the Ganoderma lucidum mycelium residue obtained from fermentation in step S4 to obtain Ganoderma lucidum mycelium residue feed with synergistic fermentation of mycelium and enzyme.

2. The method according to claim 1, characterized in that, The sieving process described in step S1 is sieving through a 40-80 mesh sieve.

3. The method according to claim 1, characterized in that, In step S2, the contents of cellulase, β-galactosidase, and xylanase were 3000 U / g, 160 U / g, and 40000 U / g, respectively.

4. The method according to claim 1, characterized in that, In step S4, the mass fractions of Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae are 8%, 8%, and 12%, respectively.

5. The method according to claim 1, characterized in that, The fermentation temperature in step S4 is 35℃, and the fermentation time is 96 hours.

6. The method according to claim 1, characterized in that, The carbon source in step S4 is corn flour.

7. Ganoderma lucidum mycelium bran prepared by the preparation method according to any one of claims 1-6.

8. The application of the Ganoderma lucidum fungus residue as described in claim 7 in the preparation of ruminant animal feed.

Images