Method for efficiently cultivating edible mushrooms through enzymolysis and fermentation of soybeans and highland barley

Abstract

The invention relates to the technical field of edible mushroom cultivation, in particular to a composite raw material for cultivating edible mushrooms through enzymolysis and fermentation of soybeans and highland barley, which is composed of the following components in parts by mass: 10-150 parts of main raw materials, 30-90 parts of composite enzyme, 0.3-8 parts of basic auxiliary materials and 0.3-8 parts of adjusting auxiliary materials: 10-150 parts of soybeans and 30-90 parts of highland barley as the main raw materials, the compound enzyme consists of amylase, protease, cellulase and lysozyme. According to the method for cultivating the edible mushrooms through enzymolysis and fermentation of the soybeans and the highland barley, the edible mushroom cultivation efficiency and the product quality are comprehensively improved, and many technical pain points of a traditional cultivation technology are solved.

Description

Technical Field

[0001] This invention relates to the field of edible fungi cultivation technology, specifically to a method for efficient cultivation of edible fungi using soybean and highland barley enzymatic fermentation. Background Technology

[0002] Edible fungi, as a food and medicinal biological resource with both nutritional and economic value, require optimized and upgraded artificial cultivation techniques for large-scale development. Currently, edible fungi cultivation often uses conventional raw materials such as sawdust, cottonseed hulls, and wheat bran in a single ratio. The large molecular nutrients in these raw materials are difficult for the fungal mycelium to absorb efficiently, resulting in low nutrient utilization and wasted nutrients in the cultivation substrate. Consequently, the yield and quality of fruiting bodies are limited.

[0003] Current cultivation techniques often employ single-enzyme hydrolysis of raw materials, resulting in limited enzyme types and fixed process parameters. This fails to fully degrade carbon and nitrogen sources in the raw materials. Furthermore, fermentation processes frequently utilize single strains and fixed temperature, humidity, and aeration conditions, leading to low nutrient conversion efficiency and susceptibility to microbial contamination, significantly reducing cultivation success rates. In addition, traditional cultivation methods lack the recycling of processed raw materials, relying solely on basic substrate for nutrition. Without precise nutrient supplementation during the fruiting period, slow mycelial growth and insufficient nutrient accumulation in fruiting bodies are common. Moreover, the cultivated mushroom fruiting bodies exhibit poor storage stability, rapid water loss, and a decline in sensory quality, failing to meet market demands for yield, quality, and shelf life. Therefore, a highly efficient mushroom cultivation method that improves nutrient utilization and optimizes the entire cultivation process is urgently needed. Summary of the Invention

[0004] The primary objective of this invention is to provide a method for the efficient cultivation of edible fungi using soybean and barley enzymatic fermentation.

[0005] A further objective of this invention is to provide a compound raw material for the enzymatic fermentation of soybeans and barley for the cultivation of edible fungi. By weight, it comprises a main raw material, a compound enzyme, basic auxiliary materials, and regulating auxiliary materials. The main raw material is soybeans and barley, with soybeans comprising 10 to 150 parts and barley comprising 30 to 90 parts. The compound enzyme comprises 0.3 to 8 parts and consists of amylase, protease, cellulase, and lysozyme. The basic auxiliary materials include 180 to 280 parts sawdust and organic auxiliary materials, which are one or more of peanut meal, corn cob, wheat bran, straw, peanut shells, cottonseed hulls, rapeseed meal, sugarcane bagasse, fungal residue, rice hulls, and traditional Chinese medicine residue. The regulating auxiliary materials comprise 4 to 9 parts lime powder and 2 to 7 parts gypsum powder.

[0006] Preferably, the amylase activity is 100 U / g to 2000 U / g, the protease activity is 3000 U / g to 30000 U / g, the cellulase activity is 8 U / g to 30 U / g, and the lysozyme activity is 300 U / g to 6000 U / g.

[0007] A method for cultivating edible fungi using soybean and highland barley through enzymatic hydrolysis and fermentation, comprising an enzymatic hydrolysis process, a fermentation process, and a cultivation process; the enzymatic hydrolysis process involves enzymatically hydrolyzing soybeans and highland barley to obtain enzymatic hydrolysis residue and enzymatic hydrolysis slurry, the enzymatic hydrolysis residue being sterilized and pulverized for later use; the fermentation process involves mixing the enzymatic hydrolysis slurry with organic auxiliary materials, sawdust, lime powder, and gypsum powder to ferment and prepare a cultivation substrate; the cultivation process involves sterilizing the cultivation substrate, inoculating it with liquid edible fungi spawn, and cultivating the fungi to obtain edible fungi through mycelial growth and fruiting; during the cultivation process, the enzymatic hydrolysis residue is used as a covering material, and the enzymatic hydrolysis slurry is used as a top dressing.

[0008] Preferably, the enzymatic hydrolysis process involves the following steps: washing and soaking soybeans and barley for 4 to 12 hours, steaming for 1 to 2 hours, adding water at a material-to-liquid ratio of 1:2 to 1:5, adjusting the temperature to 52°C to 60°C and the pH to 4.5 to 5.5, adding a compound enzyme for constant-temperature enzymatic hydrolysis for 6 to 14 hours, grinding the mixture into a slurry, boiling for 2 to 4 minutes, and separating the enzymatic residue and the enzymatic slurry.

[0009] Preferably, the enzymatic hydrolysis process adopts a stepwise enzymatic hydrolysis mode, first adding amylase and protease for 6 hours of enzymatic hydrolysis, then adding cellulase and lysozyme for 5 hours of enzymatic hydrolysis, controlling the temperature at 58℃ throughout the enzymatic hydrolysis process, adjusting the pH to 5.2 in the early stage and 4.8 in the later stage, and concentrating the enzymatic hydrolysate at 60℃ to 60% of the original volume to obtain concentrated enzymatic hydrolysate.

[0010] Preferably, the fermentation process involves the following steps: pre-wetting the enzymatic hydrolysate with organic additives for 2 to 4 hours, adding sawdust, lime powder, and gypsum powder and stirring evenly, controlling the moisture content of the mixture to be 52% to 62%, and fermenting for 48 to 144 hours using segmented temperature control, dynamic pH adjustment, and quantitative aeration. During fermentation, the mixture is turned 2 to 4 times to produce the cultivation material, with an aeration rate of 0.4 vvm to 1.2 vvm and a pH maintained at 5.8 to 7.0.

[0011] Preferably, the fermentation process incorporates a mixed fermentation strain, which is one or more of Saccharomyces cerevisiae, Aspergillus oryzae, Aspergillus niger, and Lactobacillus plantarum, with an inoculation amount of 4% to 5%; the segmented temperature control is a two-stage or three-stage temperature control, wherein the three-stage temperature control is 35°C for starting fermentation, 48°C for constant temperature fermentation, and 42°C for finishing fermentation.

[0012] Preferably, the cultivation process involves the following steps: sterilizing the cultivation material at 112°C to 121°C and 0.11MPa to 0.15MPa for 3 hours, cooling it to 23°C to 25°C, and then laying it in the planting area with a thickness of 18cm to 25cm. Inoculating the substrate with liquid edible fungi culture at a rate of 4% to 9% and covering it with a mixture of enzyme hydrolysate and powder, then allowing it to grow in the dark. After mycelial growth, controlling the temperature, humidity, and ventilation conditions for fruiting cultivation, and applying diluted enzyme hydrolysate solution during the fruiting period.

[0013] Preferably, during the mycelium growth period, the temperature is controlled at 21°C to 25°C and the humidity at 78% to 83%, with regular ventilation; during the fruiting period, the temperature is controlled at 12°C to 18°C ​​and the humidity at 88% to 95%, with increased ventilation frequency; the enzyme hydrolysis residue mixture powder is a single powder of enzyme hydrolysis residue, or a mixture of enzyme hydrolysis residue with one or more of wheat bran, wood ash, vermiculite, and perlite; the enzyme hydrolysis slurry dilution is a single dilution of enzyme hydrolysis slurry, or a mixture of enzyme hydrolysis slurry with one or more of traditional Chinese medicine extracts, amino acid foliar fertilizer, humic acid, and trace elements; the topdressing cycle during the fruiting period is 3 to 5 days.

[0014] Preferably, the edible fungus is one or more of oyster mushroom, shiitake mushroom, enoki mushroom, crab mushroom, king oyster mushroom, and white lingzhi mushroom; the cultivation process adopts a three-dimensional multi-layer planting mode, with multiple planting racks set up in the greenhouse, each layer of the planting racks spaced 40cm apart, and light control during the fruiting period, with light intensity of 300 lux to 800 lux and 4 hours of light per day.

[0015] Compared with the prior art, the beneficial effects of the present invention are: 1. The method for cultivating edible fungi by enzymatic hydrolysis and fermentation of soybeans and highland barley provided by the present invention has achieved a comprehensive improvement in the cultivation efficiency and product quality of edible fungi, and solved many technical pain points of traditional cultivation processes.

[0016] 2. This invention uses soybean and highland barley as core composite raw materials to achieve complementary advantages of carbon and nitrogen sources. It is combined with multi-component composite enzymes for enzymatic hydrolysis, which can efficiently degrade macromolecular nutrients in the raw materials and release small molecule nutrient substrates that are easily absorbed by mycelia. At the same time, lysozyme can inhibit potential contaminants in the raw materials in advance, creating a clean nutrient environment for mycelial growth.

[0017] 3. The fermentation process of this invention adopts a segmented temperature-controlled dynamic pH adjustment and a multi-strain synergistic fermentation method, which precisely matches the metabolic characteristics of the strains, greatly improves the nutrient conversion efficiency of the cultivation material, forms a stable micro-ecological system, and further reduces the probability of contamination by miscellaneous bacteria.

[0018] 4. This invention realizes the recycling of enzymatic hydrolysis products. The enzymatic hydrolysis residue can be used as a covering material to retain water and continuously release nutrients, and the enzymatic hydrolysis slurry can be used as a top dressing material to achieve precise nutrient supplementation during the fruiting period, thus constructing a closed loop of nutrient release, conversion and utilization.

[0019] 5. The precise control of temperature, humidity, ventilation, and light throughout the entire process of this invention accelerates mycelial growth, shortens the cultivation cycle, and significantly increases the content of nutrients such as protein and polysaccharides in the fruiting bodies, enhances the water retention capacity of the fruiting bodies' cells, effectively extends the storage period, and improves the sensory quality of the stored products. Detailed Implementation

[0020] 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.

[0021] Example 1:

[0022] Raw material ratio: 10 parts soybeans, 90 parts highland barley, 0.3 parts compound enzyme, 280 parts sawdust, 40 parts peanut meal, 80 parts corn cob, 4 parts lime powder, 2 parts gypsum powder. The compound enzyme consists of 100 units per gram of amylase, 3000 units per gram of protease, 8 units per gram of cellulase, and 300 units per gram of lysozyme.

[0023] Enzymatic hydrolysis process: After washing soybeans and barley, soak them for 5 hours, steam them for 1 hour, add water at a ratio of 1:2 to adjust the temperature to 52℃ and pH to 4.8, add compound enzyme and hydrolyze at a constant temperature for 6 hours, grind them into a pulp, boil for 2 minutes to separate the enzymatic residue and enzymatic pulp. After drying the enzymatic residue, sterilize it at 121℃ for 2 hours and grind it to 80 mesh for later use.

[0024] Fermentation process: After mixing enzymatic hydrolysate with peanut meal and corn cob, pre-wet and soften for 2 hours. Add sawdust, lime powder, and gypsum powder and stir thoroughly to control the moisture content to 53%. Spread the mixture into a 30 cm thick pile in the fermentation chamber. Use a two-stage temperature control: 42℃ in the early stage to promote cell growth, and 48℃ in the later stage to accelerate nutrient conversion. Check the pH every 12 hours and dynamically adjust it to 6.1 to 6.4. The aeration rate is controlled at 0.5 vvm. Fermentation lasts for 48 hours. Turn the pile twice during the fermentation period. After the pile is free of ammonia and odor, it is ready as cultivation material.

[0025] Cultivation process: Dig planting trenches 50 cm wide and 20 cm deep inside the greenhouse. Sterilize the cultivation material at 112℃ and 0.11 MPa for 3 hours, cool to 25℃, and then lay it to a thickness of 18 cm. Inoculate with 4% liquid oyster mushroom spawn. After inoculation, cover with 0.8 cm of enzyme hydrolysis residue powder. Seal the greenhouse and control the humidity at 83% and the temperature at 21℃ to prevent light from entering the mycelium. Ventilate once every 3 days during the mycelium growth period for 30 minutes each time. After the mycelium growth is completed, adjust the humidity to 88% and the temperature to 17℃. Ventilate strongly twice a day for 50 minutes each time. During the fruiting period, apply diluted enzyme hydrolysis slurry solution every 5 days at a dilution ratio of 1:8.

[0026] Example 2:

[0027] Raw material ratio: 50 parts soybeans, 50 parts highland barley, 1.2 parts compound enzyme, 250 parts sawdust, 30 parts peanut meal, 40 parts wheat bran, 100 parts straw, 60 parts peanut shells, 6 parts lime powder, 4 parts gypsum powder. The compound enzyme consists of 500 units of amylase per gram, 8000 units of protease per gram, 15 units of cellulase per gram, and 1000 units of lysozyme per gram.

[0028] Enzymatic hydrolysis process: Continuing the core technology of compound enzymatic hydrolysis in Example 1, the soaking time of soybeans and barley was adjusted to 8 hours, steaming for 2 hours, the material-to-liquid ratio was 1:4, the temperature was 60℃, the pH was 4.5, compound enzyme hydrolysis was carried out for 10 hours, and after grinding, it was boiled for 4 minutes to separate the enzymatic hydrolysis residue and the enzymatic hydrolysis pulp. The enzymatic hydrolysis residue was processed according to the method in Example 1.

[0029] Fermentation process: Continuing the two-stage temperature control logic and upgrading to a three-stage regulation: 35℃ to start fermentation, 48℃ constant temperature for 50 hours, and 42℃ to finish. Enzymatic hydrolysis pulp is mixed with peanut meal, bran, straw, and peanut shells, pre-wetted for 3 hours, and then sawdust, lime powder, and gypsum powder are added and stirred evenly. The moisture content is controlled at 58%. The pH is tested every 8 hours and dynamically maintained between 5.8 and 6.2. The aeration rate is 0.8 vvm. During the 96-hour fermentation period, the pile is turned 3 times to form a long stack of cultivation material with ventilation holes. The stack is 50 cm long, 80 cm wide at the bottom, and 18 cm high. The diameter of the ventilation holes is 2 cm and the hole spacing is 10 cm.

[0030] Cultivation process: Based on the cultivation mode of Example 1, the planting trench width is upgraded to 60 cm, the depth is 25 cm, the thickness of the cultivation material is 25 cm, the inoculation amount of liquid shiitake mushroom spawn is 6%, and the weight ratio of the enzymatic hydrolysis residue and wheat bran powder is 3:1. During the mycelium growth period, the temperature is 25℃ and the humidity is 80%, and ventilation is carried out once every 2 days for 40 minutes each time. During the fruiting period, the temperature is 15℃ and the humidity is 95%, and the ventilation frequency is increased to 4 times a day for 45 minutes each time. Topdressing is carried out with a mixture of diluted enzymatic hydrolysis pulp and Chinese herbal medicine extract, with the total proportion of mulberry leaf, dandelion and honeysuckle extract being 20%, and topdressing is carried out once every 4 days.

[0031] Example 3:

[0032] Raw material ratio: 120 parts soybeans, 30 parts highland barley, 4 parts compound enzyme, 200 parts sawdust, 150 parts cottonseed hulls, 40 parts rapeseed meal, 120 parts sugarcane bagasse, 80 parts mushroom residue, 7 parts lime powder, 5 parts gypsum powder. The compound enzyme consists of 1200 units per gram of amylase, 20000 units per gram of protease, 22 units per gram of cellulase, and 4000 units per gram of lysozyme.

[0033] Enzymatic hydrolysis process: Continuing with the enzymatic hydrolysis parameter adaptation logic of Example 2, the soaking time of soybeans and barley was adjusted to 12 hours, the cooking time was 1.5 hours, the material-to-liquid ratio was 1:5, the temperature was 55℃, the pH was 5.5, the enzymatic hydrolysis was carried out for 14 hours, the pulp was ground and boiled for 3 minutes, the enzymatic hydrolysis residue was separated, crushed and mixed with wood ash at a mass ratio of 4:1, and sterilized at 115℃ for 1.5 hours before use.

[0034] Fermentation process: Based on a three-stage control system, a multi-strain mixed fermentation culture of Saccharomyces cerevisiae, Aspergillus oryzae, and Aspergillus niger (mass ratio 1:2:1) is introduced. The inoculum is 4%. Enzymatic hydrolysate is mixed with cottonseed hulls, rapeseed meal, sugarcane bagasse, and bacterial residue. The mixture is pre-wetted for 4 hours, and sawdust, lime powder, and gypsum powder are added and stirred evenly to achieve a moisture content of 62%. During fermentation, dissolved oxygen is monitored in real time using a dissolved oxygen detector and controlled at 20% to 35% saturation. High aeration (1.2 vvm) is used in the early stage to promote cell proliferation. Medium aeration (0.7 vvm) is used in the middle stage to balance growth and metabolism. Low aeration (0.4 vvm) is used in the later stage to accumulate nutrients. The pH is adjusted in stages: growth stage 6.5 to 7.0, product stage 5.5 to 6.0. Fermentation lasts 144 hours. The pile is turned 4 times during the fermentation period. After each turning, an appropriate amount of sterile water is added to maintain the moisture content.

[0035] Cultivation process: Continuing the ventilation and temperature / humidity synergistic control logic of Example 2, the sterilization parameters for the cultivation material are 120℃, 0.15MPa for 3 hours, cooled to 23℃, and then laid to a thickness of 22 cm. Inoculate with a mixture of enoki mushrooms and crab mushrooms in liquid spawn, with an inoculation amount of 7% and a mixing ratio of 1:1. Cover with a mixture of enzymatic hydrolysis residue and wood ash powder. During the mycelial growth period, the temperature is 23℃ and the humidity is 82%, with ventilation every 2 days for 50 minutes each time. During the fruiting period, the temperature is 12℃ and the humidity is 92%, using intermittent ventilation for 30 minutes each time with a 3-hour interval. Top-dress with a mixture of amino acid foliar fertilizer and enzymatic hydrolysis slurry. The amino acid foliar fertilizer accounts for 15% of the diluted enzymatic hydrolysis slurry and is applied every 3 days.

[0036] Example 4:

[0037] Raw material ratio: 150 parts soybeans, 50 parts highland barley, 8 parts compound enzyme, 180 parts sawdust, 100 parts corn cob, 20 parts peanut meal, 150 parts rice husk, 60 parts Chinese herbal medicine residue, 9 parts lime powder, 7 parts gypsum powder. The compound enzyme consists of 2000 units per gram of amylase, 30000 units per gram of protease, 30 units per gram of cellulase, and 6000 units per gram of lysozyme.

[0038] Enzymatic hydrolysis process: Continuing the enzymatic hydrolysis system of Example 3, an innovative step-by-step enzymatic hydrolysis mode is adopted. After washing and soaking soybeans and barley for 4 hours, they are steamed for 2 hours. The material-to-liquid ratio is 1:3.5. First, amylase and protease are added for enzymatic hydrolysis for 6 hours, and then cellulase and lysozyme are added for enzymatic hydrolysis for 5 hours. The temperature is controlled at 58℃ throughout the process. The pH is adjusted to 5.2 in the early stage and 4.8 in the later stage. Stirring is performed once every 2 hours. After grinding, the concentration is increased to 60% of the original volume by low temperature at 60℃. The enzymatic hydrolysis residue and concentrated enzymatic hydrolysis pulp are separated. The enzymatic hydrolysis residue is mixed with vermiculite and perlite at a mass ratio of 2:1:1 and sterilized at 121℃ for 2 hours before use.

[0039] Fermentation process: Integrating the advantages of multi-strain mixed microorganisms and segmented regulation from Example 3, the mixed microorganisms Saccharomyces cerevisiae, Aspergillus oryzae, Lactobacillus plantarum, and Aspergillus niger were introduced in a mass ratio of 1:1:1:0.5. The inoculum amount was 5%. Concentrated enzymatic hydrolysate was mixed with corn cob, peanut meal, rice husk, and traditional Chinese medicine residue. After pre-wetting for 3 hours, sawdust, lime powder, and gypsum powder were added and stirred evenly to a moisture content of 52%. During the fermentation process, intelligent temperature control was combined with a conventional PID algorithm to adjust the temperature with an accuracy of ±0.1℃. An online pH feedback adjustment system was used to adjust the aeration rate in real time based on dissolved oxygen data, controlling it to 40% to 60% saturation. After 60 hours of fermentation, the pile was turned over again and 8% fresh sawdust and 5% cottonseed hulls were added to continue fermentation for 36 hours, during which the pile was turned over twice.

[0040] Cultivation process: Upgraded to a three-dimensional multi-layer planting mode. Three-layer planting racks are set up in the greenhouse, with each layer spaced 40 cm apart. The cultivation material is laid to a thickness of 18 cm. Inoculation with a mixed liquid spawn of king oyster mushroom and white lingzhi mushroom, with an inoculation amount of 9% and a mixing ratio of 2:1. Cover with a mixture of enzymatic hydrolysis residue, vermiculite, and perlite powder. During the mycelial growth period, the plant is protected from light and ventilated weakly twice a day for 1 hour each time. The temperature is 24℃ and the humidity is 78%. Ventilation is carried out once every 3 days for 60 minutes each time. During the fruiting period, the temperature is controlled at 18℃ and the humidity is 93%. Ventilation and light are synergistically controlled. The light intensity is 300 lux to 800 lux and 4 hours of light per day. Topdressing is done with a concentrated enzymatic hydrolysis slurry and humic acid mixed dilution solution at a dilution ratio of 1:8. After each flush of mushrooms is harvested, the enzymatic hydrolysis residue mixed powder is added to cover the plant to a thickness of 0.5 cm. At the same time, a mixture of enzymatic hydrolysis slurry and trace elements is sprayed. The trace elements account for 0.3% of the total trace elements, which are iron, zinc, and manganese complex elements.

[0041] Comparative Example 1: The raw materials are 300 parts sawdust, 200 parts cottonseed hulls, 50 parts wheat bran, 20 parts corn flour, 5 parts lime powder, and 3 parts gypsum powder. The sterilization is carried out using conventional methods at a temperature of 121℃, a pressure of 0.15MPa, and a time of 4 hours. The inoculation is 5% of the liquid spawn for oyster mushrooms. Natural mycelium growth and fruiting management are carried out without the addition of any enzymatic fermentation products. All other conditions are the same as in Example 1.

[0042] Comparative Example 2: The barley components were removed, and only 20 parts of soybeans were retained. A single amylase was used for enzymatic hydrolysis without the addition of protease, cellulase, or lysozyme. A single brewer's yeast was used for fermentation at a fixed temperature of 45℃ and a fixed pH of 6.0. The remaining raw material ratios and cultivation process were completely consistent with those in Example 1.

[0043] Comparative Example 3: 20 parts soybeans and 80 parts highland barley were washed, steamed, and then directly crushed without enzymatic hydrolysis. Fermentation was carried out using a single Aspergillus oryzae without segmented control. The remaining raw material ratios and cultivation process were the same as in Example 1.

[0044] Comparative Example 4: The cultivation process, which used 50 parts of soybeans with a single enzymatic hydrolysis and a single brewer's yeast with fixed process parameters, did not use the enzymatic hydrolysis products for topdressing and mulching. The other conditions were the same as in Example 2.

[0045] Performance testing and results analysis: Test metrics and methods: Average daily growth rate of mycelium: The length of mycelial spread 24 hours after inoculation was measured with vernier calipers. Each group had 15 replicates, and the average value was taken, in millimeters per day.

[0046] Bioconversion rate: the ratio of fresh weight of fruiting bodies to dry weight of cultivation substrate, with 10 replicates per group and the average value taken. The calculation method is: fresh weight of fruiting bodies ÷ dry weight of cultivation substrate × 100%, unit.

[0047] Fruiting body yield: The cumulative fresh weight of 4 fresh mushrooms per square meter of cultivation area, with 5 replicate plots of 10 square meters per group, and the average value is taken, in kilograms per square meter.

[0048] Nutritional composition: Protein was determined by Kjeldahl method, polysaccharide by anthrone colorimetric method, and total amino acid by high performance liquid chromatography. Each item was repeated three times and the average value was taken.

[0049] Contamination resistance rate: The percentage of cultivation area contaminated with miscellaneous bacteria is statistically analyzed. The contamination standard is the appearance of non-target mycelia or mold. Each group has 10 replicates.

[0050] Growth cycle: Record the time from inoculation to the first flush of mushrooms harvest, with 15 replicates per group, and take the average value in days.

[0051] Storage stability: After 7 days of storage at 4℃, the water loss rate and sensory evaluation (out of 10, including color, texture and odor) were determined. 20 samples were collected in each group, and the average value was taken.

[0052] The test results are shown in Table 1 below: Table 1:

[0053] The results are analyzed as follows: The performance indicators of the four embodiments of this invention are significantly better than those of the comparative examples, and show a clear progressive optimization effect. Its core mechanism and technical advantages are reflected in multi-dimensional synergistic effects. From the perspective of mycelial growth characteristics, the average daily growth rate of mycelia in Examples 1 to 4 reached 6.5 mm to 9.2 mm per day, respectively, which is up to 119.0% higher than that of the comparative examples. This improvement is due to the synergistic degradation mechanism of the compound raw materials and the compound enzyme system. The ratio of soybean to barley from 10:90 to 150:50 achieves carbon and nitrogen source complementarity. Among the compound enzymes, amylase decomposes starch into directly usable sugars, protease degrades proteins into amino acids, and cellulase breaks down plant cell walls to release bound nutrients. The three work together to provide the mycelia with comprehensive and easily absorbed nutrient substrates. Lysozyme inhibits potential contaminants in the raw materials in advance, creating a pure environment for mycelial growth. Compared with the single enzymatic hydrolysis of Comparative Example 2 and the unenzymatic treatment of Comparative Example 3, the nutrient supply efficiency has achieved a qualitative leap.

[0054] The optimization of bioconversion rate and fruiting body yield also relies on the core mechanism: the bioconversion rate of Example 4 reached 172% and the fruiting body yield reached 17.5 kg per square meter, which is 129.3% and 124.4% higher than the highest of the comparative proportions, respectively. This is the result of the synergistic effect of the entire process of enzymatic hydrolysis, fermentation and cultivation.

[0055] The temperature gradient of the staged fermentation is precisely matched to the metabolic characteristics of the mixed strains. The optimal temperature in the early stage promotes the proliferation of microorganisms such as brewer's yeast and Aspergillus oryzae. The high-oxygen environment in the middle stage accelerates nutrient conversion. The low-oxygen conditions in the later stage accumulate secondary metabolites. The diverse strains work together: brewer's yeast produces alcohol to improve the flavor of the material, Aspergillus oryzae strengthens cellulose degradation, Lactobacillus plantarum regulates the microecological balance, and Aspergillus niger synthesizes a variety of hydrolytic enzymes, forming a highly efficient nutrient conversion chain. The enzymatic residue covering layer retains water and continuously releases nutrients. The enzymatic pulp top dressing achieves precise nutrient supplementation. Compared with the non-enzymatic fermentation of Comparative Example 1 and the simplified process of Comparative Example 4, this significantly improves nutrient utilization and yield potential.

[0056] The improved nutritional composition and anti-pollution performance highlight the substantial value of the innovative mechanism: Example 4 showed a protein content of 35.6%, a polysaccharide content of 9.8%, and a total amino acid content of 24.9 grams per 100 grams, representing increases of 75.4%, 206.2%, and 99.2% respectively compared to the comparative proportions. This is due to the synergistic effect of the nutritional advantages of the compound raw materials and the biosynthetic metabolism of the multi-strain microorganisms. The high-quality protein in soybeans and the dietary fiber and trace elements in barley, after enzymatic fermentation, are partially converted into small-molecule nutrients absorbable by the mycelium of edible fungi. Furthermore, the mixed strains synthesize during the metabolic process... The enzymes are converted into polysaccharides, amino acids, and other bioactive substances, further enhancing the nutritional quality of the fruiting bodies. In terms of anti-pollution rate, all examples were controlled below 3.5%, with Example 4 at only 1.2%, a reduction of up to 93.7% compared to the comparative examples. The mechanism lies in the synergistic effect of the direct antibacterial action of lysozyme and the environmental regulation of staged fermentation. During fermentation, dynamic pH and dissolved oxygen regulation inhibit the growth of intolerant bacteria, while enzymatic hydrolysis reduces the nutrient substrates available to bacteria in the raw materials, forming a dual protection of active antibacterial action and environmental screening, effectively solving the problem of bacterial contamination that is common in the comparative examples.

[0057] The optimization of the growth cycle and storage stability demonstrates the systematic advantages of whole-process control: the growth cycle of Example 4 is only 30 days, which is 45.5% shorter than that of the comparative examples. The core mechanism is that the efficient nutrient supply and precise growth environment reduce the mycelial adaptation period and stagnation period. After 7 days of storage, the water loss rate is as low as 5.3% and the sensory score reaches 9.3 points. This is due to the sufficient accumulation of nutrients in the fruiting bodies (proteins, polysaccharides, and other components enhance the cell's water retention capacity) and the protection of the cell membrane structure by active substances in the enzymatic hydrolysis products. Compared with the high water loss rate and low sensory quality of the comparative examples, the shelf life is significantly extended. Overall, the technical solution of this invention is not a simple superposition of various links, but an organic integration of synergistic enzymatic hydrolysis of compound raw materials, segmented precise fermentation, recycling of enzymatic hydrolysis products, and whole-process parameter control. It constructs a closed-loop mechanism of nutrient release, transformation, utilization, and optimization, ultimately achieving a comprehensive improvement in cultivation efficiency, yield, quality, and stress resistance, demonstrating outstanding substantive features and significant progress.

[0058] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention.

Claims

1. A compound raw material for the enzymatic hydrolysis and fermentation of soybeans and barley for the cultivation of edible fungi, comprising, by weight parts, main raw materials, compound enzymes, basic auxiliary materials, and regulating auxiliary materials, characterized in that, The main raw materials are soybeans and highland barley, with soybeans comprising 10 to 150 parts and highland barley comprising 30 to 90 parts; the compound enzyme comprises 0.3 to 8 parts, and the compound enzyme is composed of amylase, protease, cellulase, and lysozyme; the basic auxiliary materials include 180 to 280 parts of sawdust and organic auxiliary materials, the organic auxiliary materials being one or more of peanut meal, corn cob, wheat bran, straw, peanut shells, cottonseed hulls, rapeseed meal, sugarcane bagasse, mushroom residue, rice hulls, and traditional Chinese medicine residue; the regulating auxiliary materials include 4 to 9 parts of lime powder and 2 to 7 parts of gypsum powder.

2. The composite raw material according to claim 1, characterized in that, The amylase activity is 100 U / g to 2000 U / g, the protease activity is 3000 U / g to 30000 U / g, the cellulase activity is 8 U / g to 30 U / g, and the lysozyme activity is 300 U / g to 6000 U / g.

3. A method for enzymatic fermentation of soybean and barley using the composite raw materials described in any one of claims 1 to 2 to cultivate edible fungi, characterized in that, The process includes enzymatic hydrolysis, fermentation, and cultivation. The enzymatic hydrolysis process involves treating soybeans and barley with enzymes to obtain enzymatic residue and enzymatic slurry. The enzymatic residue is sterilized and pulverized for later use. The fermentation process involves mixing the enzymatic slurry with organic additives, sawdust, lime powder, and gypsum powder to produce a cultivation substrate. The cultivation process involves sterilizing the cultivation substrate, inoculating it with liquid edible fungi spawn, and cultivating the fungi to produce fruiting bodies. During cultivation, the enzymatic residue is used as a covering material, and the enzymatic slurry is used as a top dressing.

4. The method according to claim 3, characterized in that, The enzymatic hydrolysis process is as follows: after washing soybeans and barley, soak them for 4 to 12 hours, steam them for 1 to 2 hours, add water at a material-to-liquid ratio of 1:2 to 1:5, adjust the temperature to 52°C to 60°C and the pH to 4.5 to 5.5, add a compound enzyme and hydrolyze at a constant temperature for 6 to 14 hours, grind the mixture into a pulp, boil it for 2 to 4 minutes, and separate the enzymatic residue and the enzymatic pulp.

5. The method according to claim 4, characterized in that, The enzymatic hydrolysis process adopts a stepwise enzymatic hydrolysis mode. First, amylase and protease are added for 6 hours of enzymatic hydrolysis, and then cellulase and lysozyme are added for 5 hours of enzymatic hydrolysis. The temperature is controlled at 58℃ throughout the enzymatic hydrolysis process. The pH is adjusted to 5.2 in the early stage and to 4.8 in the later stage. The enzymatic hydrolysate is concentrated to 60% of its original volume at a low temperature of 60℃ to obtain concentrated enzymatic hydrolysate.

6. The method according to claim 3, characterized in that, The fermentation process involves the following steps: pre-wetting the enzymatic hydrolysate with organic additives for 2 to 4 hours, adding sawdust, lime powder, and gypsum powder and stirring evenly, controlling the moisture content of the mixture to be 52% to 62%, and fermenting for 48 to 144 hours using segmented temperature control, dynamic pH adjustment, and quantitative aeration. During fermentation, the mixture is turned 2 to 4 times to produce the cultivation material, with an aeration rate of 0.4 vvm to 1.2 vvm and a pH maintained at 5.8 to 7.

0.

7. The method according to claim 6, characterized in that, The fermentation process involves introducing a mixed fermentation strain, which is one or more of the following: brewer's yeast, Aspergillus oryzae, Aspergillus niger, and Lactobacillus plantarum. The inoculation amount of the strain is 4% to 5%. The segmented temperature control is either a two-stage or a three-stage temperature control. The three-stage temperature control involves starting fermentation at 35°C, constant temperature fermentation at 48°C, and ending fermentation at 42°C.

8. The method according to claim 3, characterized in that, The cultivation process involves the following steps: sterilizing the cultivation material at 112℃ to 121℃ and 0.11MPa to 0.15MPa for 3 hours, cooling it to 23℃ to 25℃, and then laying it in the planting area with a thickness of 18cm to 25cm. Inoculating it with liquid edible fungi spawn at an inoculation rate of 4% to 9%, covering it with a mixture of enzyme hydrolysate and powder, and then allowing it to grow in the dark. After the mycelium has grown, the temperature, humidity, and ventilation conditions are adjusted for fruiting cultivation. During the fruiting period, diluted enzyme hydrolysate is applied as a top dressing.

9. The method according to claim 8, characterized in that, During the mycelium growth period, the temperature should be controlled at 21℃ to 25℃ and the humidity at 78% to 83%, with regular ventilation. During the fruiting period, the temperature should be controlled at 12℃ to 18℃ and the humidity at 88% to 95%, with increased ventilation frequency. The enzyme hydrolysis residue mixture powder is either a single powder of enzyme hydrolysis residue or a mixture of enzyme hydrolysis residue with one or more of wheat bran, wood ash, vermiculite, and perlite. The enzyme hydrolysis slurry dilution is either a single dilution of enzyme hydrolysis slurry or a mixture of enzyme hydrolysis slurry with one or more of traditional Chinese medicine extracts, amino acid foliar fertilizer, humic acid, and trace elements. The topdressing cycle during the fruiting period is 3 to 5 days.

10. The method according to claim 3, characterized in that, The edible fungi are one or more of the following: oyster mushrooms, shiitake mushrooms, enoki mushrooms, crab mushrooms, king oyster mushrooms, and white lingzhi mushrooms; the cultivation process adopts a three-dimensional multi-layer planting mode, with multiple planting racks set up in the greenhouse, and the spacing between each layer of the planting racks is 40cm. During the fruiting period, the light intensity is controlled in conjunction with the light regulation, with the light intensity being 300 lux to 800 lux and 4 hours of light per day.

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