Composite microbial enzyme synergistic fermented feed and preparation method thereof

Abstract

This invention discloses a compound microbial enzyme synergistic fermented feed and its preparation method, belonging to the field of animal feed production technology. The compound microbial enzyme synergistic fermented feed comprises corn, soybean meal, wheat bran, extruded flaxseed, compound microbial agents, multi-effect enzymatic hydrolysants, auxiliary additives, and pH-responsive coating agents. The microbial enzyme synergistic fermented feed of this invention has a high crude protein content and a low crude fiber content, making it easier for animals to digest and absorb. This invention significantly shortens the feed production cycle by optimizing raw material pretreatment methods, optimizing the microbial enzyme combination, and improving the fermentation process, thereby achieving synergistic effects of microorganisms and enzymes and rationally controlling the two-stage fermentation conditions.

Description

Technical Field

[0001] This invention discloses a compound bacterial enzyme synergistic fermentation feed and its preparation method, belonging to the field of animal feed production technology. Background Technology

[0002] With increasing emphasis on food safety and the full implementation of antibiotic-free farming policies, microbial-enzyme co-fermentation feed has become a core direction for improving the quality and efficiency of livestock farming. Microbial-enzyme fermented feed uses microorganisms and compound enzymes as fermentation agents to transform feed raw materials into a bio-enzyme feed that integrates microbial cell protein, bioactive small peptides and amino acids, active probiotics, and compound enzyme preparations. After fermentation, the feed becomes homogeneous and fluffy, improving palatability, stimulating animals to increase feed intake, and helping livestock grow rapidly to market weight, thus improving economic benefits. However, existing microbial-enzyme feed production methods still have many technical defects, restricting their large-scale application. 1. Low Synergistic Efficiency of Bacterial-Enzyme Combinations: Existing technologies often employ combinations of "Bacillus subtilis-Saccharomyces cerevisiae" and "cellulase-protease-xylanase," leading to substrate competition among bacterial species and overlapping enzyme targets. This results in a crude fiber degradation rate of less than 50% and limited protein hydrolysis efficiency. Furthermore, the acid production by lactic acid bacteria easily inhibits the proliferation of Bacillus subtilis, failing to fully realize the synergistic effect of bacteria and enzymes. Existing technology CN103355488B discloses a livestock and poultry feed additive, characterized by being made from the following raw materials in specific weight fractions: Astragalus membranaceus, Angelica sinensis, Glycyrrhiza uralensis, Leonurus japonicus, Codonopsis pilosula, Eucommia ulmoides, black sesame, ginseng leaf, Cuscuta chinensis, Ligustrum lucidum, Bacillus subtilis, Lactobacillus acidophilus, and Saccharomyces cerevisiae, formulated according to a certain weight ratio. However, this technology does not consider the differences in metabolic types among bacterial species, resulting in inhibited anaerobic bacterial proliferation, insufficient enzyme production efficiency of aerobic bacteria, and incomplete degradation of anti-nutritional factors. This leads to a low crude fiber degradation rate and easily causes diarrhea in young livestock.

[0003] 2. Low utilization rate of unconventional raw materials: Agricultural by-products such as stevia residue, palm meal, and flaxseed are rich in nutrients and have the potential for use in feed. However, due to their high content of anti-nutritional factors such as lignin and mannan, existing processing methods are costly, resulting in a feed addition ratio of less than 8%. This not only wastes resources but also increases environmental pressure. Existing technology with publication number CN101248835A discloses a method for enzymatic degradation of palm meal and palm kernel meal. This method mainly involves mixing dried palm meal or palm kernel meal with an enzyme solution containing mannanase and placing it at a temperature of 40–100°C for enzymatic hydrolysis. The feed raw material is obtained after 80 hours. However, this method requires pretreatment of palm meal using specialized equipment and specific processes, resulting in a long production cycle, high energy consumption, and high cost.

[0004] In view of the shortcomings of the existing technologies, developing a method for preparing compound bacterial enzyme synergistic fermentation feed with low production cost and high nutritional value has become an urgent technical problem to be solved. Summary of the Invention

[0005] To address the aforementioned problems in the existing technology, this invention provides a compound microbial enzyme synergistic fermented feed and its preparation method. By optimizing the raw material pretreatment method, optimizing the microbial enzyme combination, and improving the fermentation process, the invention achieves the objective of preparing a microbial enzyme synergistic fermented feed with a short production cycle, high nutritional value, and easy absorption by animals.

[0006] To achieve the above objectives, the following technical solution is adopted: A compound microbial enzyme synergistic fermented feed comprises, by weight, the following ingredients: 35-45 parts corn, 20-28 parts soybean meal, 6-9 parts wheat bran, 14-17 parts extruded flaxseed, 0.2-0.6 parts compound microbial agent, 0.04-0.1 parts multi-effect enzymatic hydrolysant, 0.25-0.35 parts auxiliary additives, and 1.2-2.5 parts pH-responsive coating agent.

[0007] Furthermore, the compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii.

[0008] Furthermore, the Lactobacillus reuteri has the accession number ACCC 03959 and a total viable count ≥1.2×10⁻⁶. 9 CFU / g.

[0009] Furthermore, the *Lactobacillus plantarum* has the accession number ACCC 11118 and a total viable count ≥1.2 × 10⁻⁶. 9 CFU / g.

[0010] Furthermore, the Bacillus amyloliquefaciens has the accession number ACCC 19746 and a total viable count ≥1.2 × 10⁻⁶. 9 CFU / g.

[0011] Furthermore, the Kluyveromyces martensii strain has the accession number ACCC 21190 and a total viable count ≥1.2 × 10⁻⁶. 9 CFU / g.

[0012] Furthermore, the pleotropic enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase.

[0013] Furthermore, the mannanase has an enzyme activity ≥8000 U / g.

[0014] Furthermore, the β-glucosidase has an enzyme activity ≥10000 U / g.

[0015] Furthermore, the transglutaminase has an enzyme activity ≥5000 U / g.

[0016] Furthermore, the pectinase has an enzyme activity ≥6000 U / g.

[0017] Furthermore, the α-galactosidase has an enzyme activity ≥6000 U / g.

[0018] Furthermore, the auxiliary additive is obtained by compounding L-glutamine, taurine, and tea polyphenols in a mass ratio of (2~3):(1~2):1.

[0019] Furthermore, the pH-responsive coating agent is obtained by compounding trehalose and sodium alginate in a weight ratio of (2~4):(1~3).

[0020] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are crushed and mixed with water. The mixture is then extruded and puffed before being steam-exploded to obtain a pre-treated material.

[0021] The amount of water used is 10-15% of the total weight of the raw materials.

[0022] The extrusion puffing process is carried out at a temperature of 110~125℃.

[0023] The steam explosion is described as follows: steam pressure 1.0~1.3MPa, pressure holding time 20~35s, and pressure release time <0.5s.

[0024] The weight ratio of each raw material is as follows: corn 35-45 parts, soybean meal 20-28 parts, wheat bran 6-9 parts, and puffed flaxseed 14-17 parts.

[0025] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material is put into an enzymatic hydrolysis tank, sterile water is added, the pH of the system is adjusted to 5.0~6.5, a multi-effect enzymatic hydrolysant is added and mixed evenly, and the mixture is kept warm for enzymatic hydrolysis to obtain the enzymatically hydrolyzed material.

[0026] The enzymatic hydrolysis is carried out at a controlled temperature of 43~51℃ for 5~7 hours.

[0027] The amount of sterile purified water used is 15-20% of the mass of the pretreated material.

[0028] The pH adjuster of the system is a citrate-disodium hydrogen phosphate buffer solution, which is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution; the volume ratio of 0.2M disodium hydrogen phosphate solution to 0.1M citric acid solution is 3:4.

[0029] The dosage of the multi-effect enzymatic hydrolysant is 0.04~0.1 parts.

[0030] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with the mass ratio of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase being (3~4):(2~3):(2~3):(1~2):(2~3).

[0031] Step 3: Segmented fermentation The enzymatically hydrolyzed material is transferred to a fermenter, a compound microbial agent is added, and microaerobic fermentation is carried out after the gas is replaced. After the microaerobic fermentation is completed, the gas is replaced again for anaerobic fermentation to obtain the fermented material.

[0032] During the microaerobic fermentation process, the oxygen content in the fermenter is 7% to 10%.

[0033] The microaerobic fermentation is carried out at a pressure of 0.02~0.03MPa, a temperature of 28~35℃, and a fermentation time of 6~8h.

[0034] During the anaerobic fermentation process, the oxygen content in the fermenter is <0.5%.

[0035] The anaerobic fermentation is carried out at a pressure of 0.02~0.03MPa, a temperature of 36~39℃, and a fermentation time of 8~10h.

[0036] The dosage of the compound microbial agent is 0.2 to 0.6 parts.

[0037] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with the mass ratio of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii being (2~3):(3~4):(1~2):(1~2).

[0038] The replacement gas is a mixture of N2 and CO2, wherein the volume ratio of N2 to CO2 is 9:1.

[0039] Step 4: Coating and granulation Additives are added to the fermentation feed, which is then vacuum dried and pulverized to obtain a dried feed. A pH-responsive coating agent is prepared into a coating solution, sprayed onto the surface of the dried feed, and air-dried to obtain a compound microbial enzyme synergistic fermentation feed.

[0040] The auxiliary additive is prepared by L-glutamine, taurine, and tea polyphenols in a mass ratio of (2~3):(1~2):1.

[0041] The dosage of the auxiliary additive is 0.25~0.35 parts.

[0042] The vacuum drying process involves drying at a temperature of 45~50℃.

[0043] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of (2~4):(1~3).

[0044] The amount of the pH-responsive coating agent is 1.2 to 2.5 parts.

[0045] The coating solution is prepared by dissolving a pH-responsive coating agent in pure water, and the mass concentration of the coating solution is 10-15%.

[0046] Air drying: air dry until the moisture content of the material is <10%.

[0047] The beneficial effects of this invention are as follows: 1. The compound bacterial enzyme synergistic fermented feed prepared by this invention has a crude protein content of 22.5%~25.6% and a crude fiber content of 17.1%~19.5%. The high crude protein content and low crude fiber content make it easier for animals to digest and absorb, thus improving the problem of diarrhea in animals.

[0048] 2. This invention employs a pretreatment method of extrusion puffing-steam explosion, which effectively destroys the cell walls of raw materials. Through the synergistic effect of multi-effect enzymatic hydrolysants and compound microbial agents, the fermentation conditions in the two stages are rationally controlled, effectively improving fermentation efficiency and significantly shortening the production cycle. Detailed Implementation

[0049] To make the objectives, technical solutions, and advantages of the present invention clearer, the embodiments of the present invention will be described in further detail below. It should be understood that the specific embodiments described herein are for illustrative and explanatory purposes only and are not intended to limit the scope of the invention.

[0050] Example 1: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, 35 parts corn, 25 parts soybean meal, 6 parts wheat bran, 15 parts extruded flaxseed, 0.2 parts compound microbial agent, 0.04 parts multi-effect enzymatic hydrolysant, 0.3 parts auxiliary additives, and 2 parts pH-responsive coating agent.

[0051] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water is then evenly sprayed onto the mixture using a spraying device, with the amount of pure water being 15% of the raw material weight. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 110℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.0 MPa and held for 35 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0052] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 15% of the mass of the pretreated material. The pH of the system was adjusted to 5.0 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 45°C, and 0.04 parts of pleuronectin hydrolysant were added and mixed evenly. The mixture was kept at this temperature for 6 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0053] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0054] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 3:2:2:1:2.

[0055] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.2 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 7% for microaerobic fermentation. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.02 MPa, and the temperature was adjusted to 28℃ for 8 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5% for anaerobic fermentation. The temperature was adjusted to 38℃ for 8 hours of fermentation to obtain the fermented material.

[0056] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 2:3:2:1.

[0057] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 45℃ until the moisture content is 15%, then pulverize to 80 mesh to obtain dried material. Dissolve the pH-responsive coating agent in pure water to prepare a 10% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 55℃ and the outlet air temperature to 40℃. After the coating solution is sprayed, continue to air dry in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 2:1:1.

[0058] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 3:2.

[0059] Example 2: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, 35 parts corn, 26 parts soybean meal, 9 parts wheat bran, 16 parts extruded flaxseed, 0.6 parts compound microbial agent, 0.07 parts multi-effect enzymatic hydrolysant, 0.3 parts auxiliary additives, and 1.5 parts pH-responsive coating agent.

[0060] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water is then evenly sprayed onto the mixture using a spraying device, with the amount of pure water being 15% of the raw material weight. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 120℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.3 MPa and held for 20 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0061] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 15% of the mass of the pretreated material. The pH of the system was adjusted to 6.0 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 45°C, and 0.07 parts of pleuronectin hydrolysant were added and mixed evenly. The mixture was kept at this temperature for 6 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0062] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0063] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 4:3:3:2:3.

[0064] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.6 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 7%, and microaerobic fermentation was carried out. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.03 MPa, and the temperature was adjusted to 35℃ for 7 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5%, and anaerobic fermentation was carried out. The temperature was adjusted to 37℃ for 9 hours of fermentation to obtain the fermented material.

[0065] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 2:3:2:1.

[0066] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 45℃ until the moisture content is 10%, then pulverize to 60 mesh to obtain dried material. Dissolve the pH-responsive coating agent in pure water to prepare a 10% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 55℃ and the outlet air temperature to 45℃. After the coating solution is sprayed, continue to air dry in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 3:1:1.

[0067] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 4:3.

[0068] Example 3: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, 45 parts corn, 20 parts soybean meal, 9 parts wheat bran, 17 parts extruded flaxseed, 0.5 parts compound microbial agent, 0.1 parts multi-effect enzymatic hydrolysant, 0.25 parts auxiliary additives, and 1.2 parts pH-responsive coating agent.

[0069] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water (10% of the raw material weight) is then evenly sprayed and mixed using a spraying device. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 120℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.3 MPa and held for 20 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0070] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 20% of the mass of the pretreated material. The pH of the system was adjusted to 5.0 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 43°C, and 0.1 parts of pleuronectin hydrolysant were added and mixed evenly. The mixture was kept at this temperature for 6 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0071] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0072] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 3:2:2:2:3.

[0073] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.5 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 10% for microaerobic fermentation. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.02 MPa, and the temperature was adjusted to 30℃ for 6 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5% for anaerobic fermentation. The temperature was adjusted to 39℃ for 8 hours of fermentation to obtain the fermented material.

[0074] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 2:3:2:2.

[0075] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 45℃ until the moisture content is 10%, then pulverize to 60 mesh to obtain dried material. Dissolve the pH-responsive coating agent in pure water to prepare a 10% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 54℃ and the outlet air temperature to 40℃. After the coating solution is sprayed, continue to air dry in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 3:2:1.

[0076] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 3.5:2.

[0077] Example 4: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, the following ingredients: 40 parts corn, 28 parts soybean meal, 6 parts wheat bran, 14 parts extruded flaxseed, 0.3 parts compound microbial agent, 0.05 parts multi-effect enzymatic hydrolysant, 0.35 parts auxiliary additives, and 1.5 parts pH-responsive coating agent.

[0078] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water (10% of the raw material weight) is then evenly sprayed and mixed using a spraying device. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 125℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.0 MPa and held for 30 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0079] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 15% of the mass of the pretreated material. The pH of the system was adjusted to 6.0 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 50°C, and 0.05 parts of pleuronectin hydrolysant were added and mixed evenly. The mixture was kept at this temperature for 5 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0080] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0081] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 4:3:3:1:2.

[0082] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.3 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 7%, and microaerobic fermentation was carried out. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.02 MPa, and the temperature was adjusted to 35℃ for 7 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5%, and anaerobic fermentation was carried out. The temperature was adjusted to 38℃ for 8 hours of fermentation to obtain the fermented material.

[0083] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 3:4:1:2.

[0084] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 50℃ until the moisture content is 10%, then pulverize to 80 mesh to obtain dried material. Dissolve a pH-responsive coating agent in pure water to prepare a 15% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 54℃ and the outlet air temperature to 45℃. After the coating solution is sprayed, continue air drying in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 2:1:1.

[0085] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 2:1.

[0086] Example 5: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, the following ingredients: 40 parts corn, 25 parts soybean meal, 6 parts wheat bran, 15 parts extruded flaxseed, 0.5 parts compound microbial agent, 0.08 parts multi-effect enzymatic hydrolysant, 0.25 parts auxiliary additives, and 2.5 parts pH-responsive coating agent.

[0087] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water (10% of the raw material weight) is then evenly sprayed and mixed using a spraying device. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 125℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.0 MPa and held for 30 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0088] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 20% of the mass of the pretreated material. The pH of the system was adjusted to 6.0 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 51°C, and 0.08 parts of pleuronectin hydrolysant were added and mixed evenly. The mixture was kept at this temperature for 6 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0089] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0090] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 3:2:3:1:2.

[0091] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.5 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 7% for microaerobic fermentation. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.02 MPa, and the temperature was adjusted to 35℃ for 6 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5% for anaerobic fermentation. The temperature was adjusted to 39℃ for 8 hours of fermentation to obtain the fermented material.

[0092] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 2:3:2:1.

[0093] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 45℃ until the moisture content is 15%, then pulverize to 60 mesh to obtain dried material. Dissolve the pH-responsive coating agent in pure water to prepare a 10% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 58℃ and the outlet air temperature to 45℃. After the coating solution is sprayed, continue to air dry in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 3:1:1.

[0094] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 2:1.5.

[0095] Example 6: A compound microbial enzyme synergistic fermentation feed A compound microbial enzyme synergistic fermented feed comprises, by weight, 45 parts corn, 20 parts soybean meal, 9 parts wheat bran, 14 parts extruded flaxseed, 0.6 parts compound microbial agent, 0.08 parts multi-effect enzymatic hydrolysant, 0.3 parts auxiliary additives, and 2 parts pH-responsive coating agent.

[0096] A method for preparing a compound microbial enzyme synergistic fermentation feed includes the following steps: Step 1: Raw material pretreatment Corn, soybean meal, wheat bran, and puffed flaxseed are pulverized to below 20 mesh using a pulverizer. Pure water is then evenly sprayed onto the mixture using a spraying device, with the amount of pure water being 15% of the raw material weight. The uniformly mixed material is fed into the feed hopper of a twin-screw extruder, with the extruder temperature controlled at 110℃. After discharge from the twin-screw extruder, the material is sent to a steam explosion device. The steam pressure is set to 1.0 MPa and held for 30 seconds. Depressurization is then performed using an instantaneous depressurization method, controlling the depressurization time to <0.5 seconds. After depressurization and discharge, the material is further pulverized to below 30 mesh using a pulverizer to obtain pre-treated material.

[0097] Step 2: Enzymatic hydrolysis of pretreated material The pretreated material obtained in step 1 was placed into an enzymatic hydrolysis tank, and sterile purified water was added. The amount of sterile purified water was 15% of the mass of the pretreated material. The pH of the system was adjusted to 6.5 with citrate-disodium hydrogen phosphate buffer. The temperature was raised to 45°C, and 0.08 parts of pleurodesis agent were added and mixed evenly. The mixture was kept at this temperature for 7 hours to obtain the enzymatically hydrolyzed material. During the incubation process, the mixture was stirred for 1 minute every 1.5 hours.

[0098] The citrate-disodium hydrogen phosphate buffer solution is prepared by mixing 0.2M disodium hydrogen phosphate solution and 0.1M citric acid solution, with a volume ratio of 3:4.

[0099] The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with a mass ratio of 4:3:2:2:3.

[0100] Step 3: Segmented fermentation The enzymatically hydrolyzed material obtained in step 2 was transferred to a fermenter, and 0.6 parts of compound bacterial agent were added and stirred evenly. The air inside the fermenter was replaced with a N2-CO2 mixed gas until the oxygen content reached 7% for microaerobic fermentation. The pressure inside the fermenter was maintained at a slightly positive pressure of 0.03 MPa, and the temperature was adjusted to 30℃ for 7 hours of fermentation. Then, the air inside the fermenter was replaced with an N2-CO2 mixed gas until the oxygen content was <0.5% for anaerobic fermentation. The temperature was adjusted to 36℃ for 10 hours of fermentation to obtain the fermented material.

[0101] The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with a mass ratio of 2:3:1:2.

[0102] Step 4: Coating and granulation Add auxiliary additives to the fermentation material obtained in step 3, stir and mix well, vacuum dry at 50℃ until the moisture content is 10%, then pulverize to 60 mesh to obtain dried material. Dissolve the pH-responsive coating agent in pure water to prepare a 10% (w / w) coating solution. Feed the dried material into a fluidized bed coating machine, controlling the atomization pressure to 0.25 MPa to ensure the coating solution is evenly sprayed onto the surface of the dried material. Control the inlet air temperature of the fluidized bed coating machine to 55℃ and the outlet air temperature to 40℃. After the coating solution is sprayed, continue air drying in the fluidized bed coating machine until the material moisture content is <10%. Cool the discharged material to room temperature to obtain a compound microbial enzyme synergistic fermented feed. The auxiliary additive is formulated from L-glutamine, taurine, and tea polyphenols in a mass ratio of 2:1:1.

[0103] The pH-responsive coating agent is prepared by mixing trehalose and sodium alginate in a mass ratio of 3:2.

[0104] Performance testing 1. The crude fiber and crude protein content of the compound bacterial enzyme co-fermentation feeds in Examples 1-6 were detected by the following methods; the results are shown in Table 1.

[0105] Crude fiber content: The crude fiber content in the feed was determined according to GB / T 6434-2022 "Determination of Crude Fiber Content in Feed - Filtration Method"; Crude protein content: The crude protein content in the feed was determined according to GB / T 6432-2018 "Determination of crude protein in feed - Kjeldahl method". Table 1. Results of crude fiber and crude protein content detection in feed co-fermented with compound microorganisms and enzymes. Analysis of the test results in Table 1 shows that the compound bacterial enzyme synergistic fermented feed prepared by the present invention has a crude protein content of 22.5%~25.6% and a crude fiber content of 17.1%~19.5%, indicating that the fermented feed prepared by the present invention has a high crude protein content and its preparation method has a higher crude fiber degradation efficiency.

[0106] 2. Animal production performance: (1) Piglet feeding experiment: 220 weaned piglets aged 21 days were randomly divided into 4 groups: experimental group 1, experimental group 2, experimental group 3, and control group 1, with 55 piglets in each group; experimental group 1 was fed the feed of Example 1, experimental group 2 was fed the feed of Example 2, experimental group 3 was fed the feed of Example 3, and control group 1 was fed commercially available ordinary fermented feed for piglets; the experiment lasted for 48 days, and the daily weight gain and diarrhea rate were recorded; and the feed conversion ratio was calculated based on the daily feed intake. The experimental results are shown in Table 2.

[0107] Table 2 Results of piglet feeding trial Analysis of the test results in Table 2 shows that, compared with piglets fed commercial feed, piglets fed the fermented feed of this invention have a lower diarrhea rate, only 3.63%~5.45%; the average daily weight gain of piglets is 485g~535g, and the feed conversion ratio is 1.94~2.03.

[0108] In summary, in animal experiments, animals fed with the fermented feed of this invention were less prone to diarrhea, grew faster, and had a lower feed conversion ratio, which helped save on breeding costs.

[0109] Obviously, there are many other possible implementation methods under the concept of this invention. It should be stated here that any changes made under the inventive concept of this invention will fall within the protection scope of this invention.

Claims

1. A method for preparing a compound microbial enzyme synergistic fermentation feed, characterized in that: Includes the following steps: Step 1: Crush corn, soybean meal, wheat bran, and puffed flaxseed and mix them with water. Extrude and puff the mixture and then steam-explode it to obtain the pre-treated material. The weight ratio of each raw material is as follows: corn 35-45 parts, soybean meal 20-28 parts, wheat bran 6-9 parts, and puffed flaxseed 14-17 parts; Step 2: Put the pretreated material into the enzymatic hydrolysis tank, add sterile water, adjust the pH of the system to 5.0~6.5, add multi-effect enzymatic hydrolysant, mix evenly, keep warm and enzymatically hydrolyze to obtain enzymatically hydrolyzed material; The multi-effect enzymatic hydrolysate is composed of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase, with the mass ratio of mannanase, β-glucosidase, transglutaminase, pectinase, and α-galactosidase being (3~4):(2~3):(2~3):(1~2):(2~3). Step 3: Transfer the enzymatically hydrolyzed material into a fermenter, add compound microbial agents, replace the gas, and carry out microaerobic fermentation; after the microaerobic fermentation is completed, replace the gas again and carry out anaerobic fermentation to obtain fermented material; The compound microbial agent is composed of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii, with the mass ratio of Lactobacillus reuteri, Lactobacillus plantarum, Bacillus amyloliquefaciens, and Kluyveromyces martensii being (2~3):(3~4):(1~2):(1~2). Step 4: Add auxiliary additives to the fermentation material, vacuum dry and then pulverize to obtain dried material. Prepare a pH-responsive coating agent into a coating solution, spray it onto the surface of the dried material, and air dry to obtain a compound microbial enzyme synergistic fermentation feed; The auxiliary additive is prepared by L-glutamine, taurine, and tea polyphenols in a mass ratio of (2~3):(1~2):

1.

2. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The amount of water used in step 1 is 10-15% of the total weight of the raw materials.

3. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The extrusion puffing in step 1: extrusion puffing temperature 110~125℃; the steam explosion: steam pressure 1.0~1.3MPa, pressure holding 20~35s, pressure release time <0.5s.

4. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The dosage of the multi-effect enzymatic hydrolysant in step 2 is 0.04~0.1 parts; the incubation enzymatic hydrolysis is carried out at a controlled temperature of 43~51℃ for 5~7 hours.

5. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The amount of sterile water used in step 2 is 15-20% of the mass of the pretreated material.

6. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The dosage of the compound microbial agent in step 3 is 0.2 to 0.6 parts.

7. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: Step 3: Microaerobic fermentation: internal pressure 0.02~0.03MPa, temperature 28~35℃, fermentation time 6~8h; during microaerobic fermentation, the oxygen content in the fermentation tank is 7%~10%.

8. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The anaerobic fermentation described in step 3: internal pressure 0.02~0.03MPa, temperature 36~39℃, fermentation time 8~10h.

9. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The pH-responsive coating agent described in step 4 is prepared by mixing trehalose and sodium alginate in a mass ratio of (2~4):(1~3); the amount of the pH-responsive coating agent is 1.2~2.5 parts.

10. The method for preparing a compound microbial enzyme synergistic fermentation feed as described in claim 1, characterized in that: The coating solution described in step 4 is prepared by dissolving a pH-responsive coating agent in water to form a coating solution with a mass concentration of 10-15%.

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