Mushroom compost and method of making same

Abstract

This invention provides a mushroom culture medium and its preparation method: A compound biological agent is prepared by mixing a compound enzyme preparation with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50-1:100 to obtain the biological agent. The compound enzyme preparation includes 50,000-100,000 U / g of protease, 10,000-20,000 U / g of acidic cellulase, 10,000-20,000 U / g of neutral cellulase, 50,000-100,000 U / g of xylanase, laccase, 1,000-5,000 U / g of aminopeptidase, and 10,000-20,000 U / g of glucose oxidase. The durian shells are crushed and mixed with the biological agent at a mass ratio of 3:7-1:1. Solid-state fermentation is then performed on the uniformly mixed durian shells at a fermentation temperature of 30-37℃ for 48-96 hours. After fermentation, the durian shells are used as a mushroom culture medium for mushroom cultivation, thus solving the problem of disposing of durian shells and Bacillus subtilis fermentation waste liquid.

Description

Technical Field

[0001] This invention belongs to the field of mushroom culture medium technology, specifically relating to a method for preparing mushroom culture medium by solid-state fermentation of durian shells using Bacillus subtilis fermentation waste liquid. Background Technology

[0002] Durian is a tropical fruit. Its peel is rich in protein, fat, and cellulose, and also contains folic acid, niacin, and inorganic salts. The main components of durian shell are 60.45% cellulose, 13.09% hemicellulose, and 15.45% lignin. Currently, the main methods for disposing of durian shells are incineration or landfill, which pollutes the environment and wastes the organic matter in the shells.

[0003] Mushrooms are saprophytic fungi that rely entirely on the nutrients in their substrate for growth and development. Mushrooms can utilize carbon sources such as glucose, sucrose, maltose, starch, cellulose, hemicellulose, and lignin, but these must be broken down into simple carbohydrates by other microorganisms and enzymes secreted by the mushroom mycelium before they can be absorbed and utilized. Mushrooms can utilize nitrogen sources such as urea, ammonium salts, peptone, and amino acids. In addition, mushroom growth requires certain amounts of minerals such as phosphorus, potassium, and calcium, as well as trace elements such as iron and molybdenum. Currently, the raw materials for mushroom substrate include rice straw, wheat straw, poultry and livestock manure, superphosphate, and gypsum powder. During preparation, the rice straw is dried and chopped, and the manure is dried, crushed, and then moistened with water. The mixture is layered, alternating between straw and manure, and then covered with straw mats. After 3-4 days, the pile is turned over once; if the pile is dry, appropriate amounts of water can be added. After 7-8 days, the pile is turned over a second time, and gypsum powder is added. After 11-12 days, turn the compost for the third time and add superphosphate. On the 13th day, while it is still hot, move the substrate into the mushroom house, spread it evenly on the mushroom bed, and seal the room. Then raise the indoor temperature to about 60℃ and maintain it for 2-3 hours. Then lower the temperature to 52℃ and maintain it for 4-7 days. When the temperature of the substrate drops to 28℃, start sowing. The production process requires many types of raw materials and involves complex steps.

[0004] Bacillus subtilis is a common probiotic widely used in animal husbandry and plant cultivation. Currently, the preparation of Bacillus subtilis mainly relies on high-density liquid fermentation. During production, the fermentation broth undergoes solid-liquid separation and further purification to produce the final product. This process generates a large amount of fermentation waste liquid. During fermentation, Bacillus subtilis secretes large amounts of digestive enzymes, bacteriostatic agents, γ-polyglutamic acid, and other substances, along with residual trace elements from the culture medium and a small amount of unseparated bacterial cells. These substances accumulate in the supernatant. Direct discharge of this waste liquid along with the fermentation waste liquid causes both environmental pollution and resource waste.

[0005] The research direction of this invention is to combine these three elements to fully utilize the useful components in durian shells and fermentation waste liquid, thereby avoiding environmental pollution and transforming them into products with market value. Summary of the Invention

[0006] Based on the above description, the present invention provides a method for preparing mushroom culture medium by solid-state fermentation of Bacillus subtilis fermentation waste liquid and durian shells together. This method not only solves the problem of disposing of durian shells and Bacillus subtilis fermentation waste liquid, but also transforms environmentally polluting waste into products that can be sold on the market.

[0007] This invention first provides a method for preparing mushroom culture medium, and the technical solution adopted to solve its technical problem is as follows:

[0008] A method for preparing mushroom culture medium.

[0009] S1: Compound biological agent; The compound enzyme preparation is compounded with Bacillus subtilis fermentation waste liquid to obtain the biological agent;

[0010] S2: Mixing; Crush the durian shell and mix it evenly with the biological agent;

[0011] S3: Solid-state fermentation; The durian shells, after being mixed evenly, are fermented in a solid state. After fermentation, the durian shells are used as a mushroom culture medium for mushroom cultivation.

[0012] Based on the above technical solution, the present invention can be further improved as follows.

[0013] Furthermore, in the above-mentioned method for preparing a mushroom culture medium, the compound enzyme preparation includes: protease, acidic cellulase, neutral cellulase, xylanase, laccase, aminopeptidase, and glucose oxidase.

[0014] Furthermore, in the above-mentioned method for preparing a mushroom culture medium, the compound enzyme preparation formula comprises: protease 50,000-100,000 U / g, acidic cellulase 10,000-20,000 U / g, neutral cellulase 10,000-20,000 U / g, xylanase 50,000-100,000 U / g, laccase 1,000-2,000 U / g, aminopeptidase 1,000-5,000 U / g, and glucose oxidase 10,000-20,000 U / g.

[0015] Furthermore, in the above-mentioned method for preparing a mushroom culture medium, the compound enzyme preparation is mixed with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50-1:100 to obtain a biological agent.

[0016] Furthermore, in the above-mentioned method for preparing a mushroom culture medium, the durian shell and the biological agent are mixed evenly in a mass ratio of 3:7 to 1:1.

[0017] Furthermore, in the above-mentioned method for preparing a mushroom culture medium, in step S3, the fermentation temperature is 30-37℃ and the fermentation time is 48-96h.

[0018] The present invention also provides a mushroom culture medium prepared according to any one of the above preparation methods.

[0019] Furthermore, the mushroom culture medium described above produces mushrooms with three fruiting flushes.

[0020] Furthermore, in the aforementioned mushroom culture medium, the mushroom is oyster mushroom, king oyster mushroom, or shimeji mushroom.

[0021] Furthermore, when the above-mentioned mushroom culture medium is used to cultivate oyster mushrooms, the mushroom yield ratio is 0.90 to 1.05.

[0022] Compared with the prior art, the technical solution of the present invention has the following beneficial technical effects:

[0023] This invention provides a method for producing mushroom culture medium by co-fermenting Bacillus subtilis fermentation waste liquid with durian shells. The biological agent utilizes the Bacillus subtilis fermentation waste liquid to ferment durian shells, and the fermented durian shells are then used as mushroom culture medium. This fully utilizes both the fermentation waste liquid and durian shells, achieving waste utilization and reducing environmental pollution. Simultaneously, it fully utilizes the useful components in both durian shells and fermentation waste liquid, improving mushroom yield and quality.

[0024] Cellulase promotes the degradation of cellulose, hemicellulase hydrolyzes polysaccharides (except cellulose and pectin) in plant cell walls, and laccase accelerates the decomposition of aromatic macromolecules such as lignin. The synergistic effect of cellulase and laccase can accelerate the degradation of plant cellulose and lignin. The main function of proteases is to promote the degradation of proteins into small peptides. Aminopeptidases are exoproteases that can cleave amino acid residues from the amino terminus of proteins and polypeptides to produce free amino acids. The combined use of proteases and aminopeptidases can further promote the conversion of proteins into small peptides and amino acids. This invention utilizes Bacillus subtilis fermentation waste liquid to ferment durian shells, and combines it with laccase, cellulase, hemicellulase, xylanase, protease, and aminopeptidases to achieve a synergistic effect.

[0025] In the process of edible fungi spawn production, adding laccase, cellulase, and xylanase can accelerate the decomposition of lignin and cellulose in durian shells, providing nutrients for mycelium. At the same time, laccase can oxidize phenols into quinones, which act as toxins to resist contamination by other fungi. Adding protease and aminopeptidase can degrade proteins in durian shells and form small peptides and amino acids, providing the nitrogen source needed for rapid mycelial growth.

[0026] The durian shells of this invention can be directly used as mushroom culture medium after solid-state fermentation, without the need for other steps.

[0027] The mushroom culture medium prepared by the method of the present invention can be used directly as a complete culture medium without the need to add wheat straw, poultry and livestock manure or other ingredients. Detailed Implementation

[0028] To facilitate understanding of this application, a more complete description will be provided below. This application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that the disclosure of this application will be thorough and complete.

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0031] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0032] The enzymes used in this invention are all from Wuhan Xinhua Yang Biotechnology Co., Ltd.

[0033] A method for preparing mushroom culture medium.

[0034] S1: Compound biological agent; The compound enzyme preparation is compounded with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50-1:100 to obtain the biological agent; The compound enzyme preparation formula is as follows: protease 50000-100000U / g, acidic cellulase 10000-20000U / g, neutral cellulase 10000-20000U / g, xylanase 50000-100000U / g, laccase 1000-2000U / g, aminopeptidase 1000-5000U / g, glucose oxidase 10000-20000U / g.

[0035] S2: Mixing; Crush the durian shell and mix it evenly with the biological agent at a mass ratio of 3:7-1:1.

[0036] S3: Solid-state fermentation; The durian shells are mixed evenly and fermented in a solid state at a temperature of 30-37℃ for 48-96 hours. After fermentation, the durian shells are used as a mushroom culture medium for mushroom cultivation.

[0037] Example 1

[0038] The preparation method of mushroom culture medium is as follows: the compound enzyme preparation is added in the following proportions: protease 50000U / g, acidic cellulase 20000U / g, neutral cellulase 20000U / g, xylanase 50000U / g, laccase 2000U / g, aminopeptidase 1000U / g, and glucose oxidase 10000U / g. It is mixed with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50, and the mass ratio of durian shell to biological agent is 3:7. The fermentation temperature is 30℃, and the fermentation time is 96 hours.

[0039] Example 2

[0040] The preparation method of mushroom culture medium is as follows: the compound enzyme preparation is added in the following proportions: protease 80000U / g, acidic cellulase 15000U / g, neutral cellulase 15000U / g, xylanase 80000U / g, laccase 1500U / g, aminopeptidase 2000U / g, and glucose oxidase 15000U / g. It is mixed with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:75, and the mass ratio of durian shell to biological agent is 4:6. The fermentation temperature is 35℃, and the fermentation time is 72 hours.

[0041] Example 3

[0042] The preparation method of mushroom culture medium is as follows: the compound enzyme preparation is added in the following proportions: protease 100,000 U / g, acidic cellulase 10,000 U / g, neutral cellulase 10,000 U / g, xylanase 100,000 U / g, laccase 2,000 U / g, aminopeptidase 3,000 U / g, and glucose oxidase 20,000 U / g. It is mixed with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:100, and the mass ratio of durian shell to biological agent is 1:1. The fermentation temperature is 37℃, and the fermentation time is 48 hours.

[0043] Comparative Example 1

[0044] Comparative Example 1 was the same as Example 1 in all other conditions except that the Bacillus subtilis fermentation waste liquid was replaced with water.

[0045] Comparative Example 2

[0046] Comparative Example 2 was conducted under the same conditions as Example 2, except that the Bacillus subtilis fermentation waste liquid was replaced with water.

[0047] Comparative Example 3

[0048] Comparative Example 3 was conducted under the same conditions as Example 3, except that the Bacillus subtilis fermentation waste liquid was replaced with water.

[0049] Comparative Example 4

[0050] Comparative Example 4 was the same as Example 1 in all other conditions except that the Bacillus subtilis fermentation waste liquid was replaced with distillers' grains fermentation waste liquid.

[0051] Compare with Example 1

[0052] Comparative Example 1 is basically the same as Example 1, except that no compound enzyme preparation is added, the mass ratio of durian shell to Bacillus subtilis fermentation waste liquid is 1:1, the fermentation temperature is 37℃, and the fermentation time is 48h.

[0053] Compare with Example 2

[0054] Comparative Example 2 is basically the same as Example 1, except that no compound enzyme preparation and Bacillus subtilis fermentation waste liquid are added, the mass ratio of durian shell to water is 1:1, the fermentation temperature is 37℃, and the fermentation time is 48h.

[0055] Application Examples

[0056] Fermented durian shells were used as culture medium, bagged, and inoculated for cultivation. Ten replicates were performed per group. The growth performance of oyster mushrooms was compared based on the yield ratio. The yield ratio was calculated as: actual weight of oyster mushrooms produced (catties) / weight of durian shells used (catties). The increase in the yield ratio was used to more intuitively evaluate the growth performance of oyster mushrooms. The increase in the yield ratio was relative to Control Example 1, and was calculated as: (Yield ratio of each group - Yield ratio of Control Example 1) / Yield ratio of Control Example 1 * 100%.

[0057] Application of comparative examples

[0058] Oyster mushroom culture medium was purchased from the market, bagged and inoculated, and fermented in bags for cultivation. Each group had 10 replicates to compare the growth performance of oyster mushrooms and the increase in mushroom production ratio compared with control 1.

[0059] Table 1. Growth performance of oyster mushrooms

[0060] Group Mushroom production ratio Mushroom production ratio increase Mushroom season Application of comparative examples 0.78 25.81％ 3 Compare with Example 1 0.62 / 1 Compare with Example 2 0.58 -6.45％ 1 Comparative Example 1 0.63 1.61％ 1 Comparative Example 2 0.71 14.52％ 1 Comparative Example 3 0.68 9.68％ 2 Comparative Example 4 0.58 -6.45％ 1 Example 1 1.02 64.52％ 3 Example 2 1.05 69.35％ 3 Example 3 0.96 54.84％ 3

[0061] The results in Table 1 show that the effect of not using Bacillus subtilis fermentation waste liquid, or using only Bacillus subtilis fermentation waste liquid without adding biological agents, is not as good as using both in combination. Meanwhile, with other conditions unchanged, replacing the Bacillus subtilis fermentation waste liquid with distiller's grains fermentation waste liquid resulted in a significantly lower mushroom yield than in the embodiments of this invention.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the invention by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the invention should be included within the scope of protection of the invention.

Claims

1. A method for preparing a mushroom culture medium, characterized in that, Includes the following steps: S1: Compound biological agent; The compound enzyme preparation is compounded with Bacillus subtilis fermentation waste liquid to obtain the biological agent; S2: Mixing; Crush the durian shell and mix it evenly with the biological agent; S3: Solid-state fermentation; The durian shells, after being mixed evenly, are fermented in a solid state. After fermentation, the durian shells are used as a mushroom culture medium for mushroom cultivation. The compound enzyme preparation includes: protease, acidic cellulase, neutral cellulase, xylanase, laccase, aminopeptidase, and glucose oxidase. The compound enzyme preparation is mixed with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50-1:100 to obtain a biological agent; The durian shell and biological agent are mixed evenly in a mass ratio of 3:7 to 1:

1.

2. The method for preparing a mushroom culture medium according to claim 1, characterized in that, The compound enzyme preparation formulation consists of 50,000-100,000 U / g of protease, 10,000-20,000 U / g of acidic cellulase, 10,000-20,000 U / g of neutral cellulase, 50,000-100,000 U / g of xylanase, 1,000-2,000 U / g of laccase, 1,000-5,000 U / g of aminopeptidase, and 10,000-20,000 U / g of glucose oxidase.

3. The method for preparing a mushroom culture medium according to claim 1, characterized in that, In step S3, the fermentation temperature is 30-37℃ and the fermentation time is 48-96h.

4. Mushroom culture medium prepared by the preparation method according to any one of claims 1 to 3.

5. The mushroom culture medium according to claim 4, characterized in that, Mushrooms cultivated using this substrate produce three flushes of fruiting.

6. The mushroom culture medium according to claim 4, characterized in that, The mushrooms mentioned are oyster mushrooms, king oyster mushrooms, or shimeji mushrooms.

7. The mushroom culture medium according to claim 6, characterized in that, When oyster mushrooms are cultivated using this mushroom substrate, the yield ratio is 0.90~1.05.