Enzymolysis-fermentation synergistic biological peptide feed additive and preparation method thereof
By using an enzymatic hydrolysis-fermentation synergistic biotransformation process, combined with compound microbial agents and traditional Chinese medicine decoctions, the problem of single additives being unable to meet multiple needs has been solved, achieving efficient nutrition and immune function, and improving feed utilization and animal health.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- JIANGSU FUHAI BIOLOGICAL TECH CO LTD
- Filing Date
- 2026-03-18
- Publication Date
- 2026-05-12
AI Technical Summary
Existing single enzymatic hydrolysis or fermentation technologies are difficult to meet the multiple needs of feed additives in terms of nutrition, immunity, etc., and microbial fermentation has low efficiency, is easily contaminated by miscellaneous bacteria, and the product has limited functions.
The enzymatic hydrolysis-fermentation synergistic biotransformation process uses components such as rapeseed meal, cellulase, neutral protease, and astragalus polysaccharide, combined with a compound bacterial agent of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis, and adds traditional Chinese medicine decoction and zeolite powder to achieve multi-strain synergistic fermentation and solid-state treatment.
It improves the nutritional value and utilization efficiency of feed ingredients, enhances animal immunity and disease resistance, improves intestinal microecology, and improves the physical and storage stability of products.
Smart Images
Figure CN122004351A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of feed additive technology, specifically to an enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive and its preparation method. Background Technology
[0002] To ensure animal food safety and public health, the use of antibiotics in feed is strictly limited globally, prompting the livestock industry to demand green feed additives with functions such as growth promotion and diarrhea prevention. Soybean meal and other protein raw materials are expensive and their supply fluctuates. Rapeseed meal contains anti-nutritional factors, limiting its application and necessitating its degradation and transformation into safe, high-value feed ingredients.
[0003] Single enzymatic hydrolysis technology efficiently degrades proteins to produce peptides, but the fiber destruction is limited and the product has a single function; single microbial fermentation technology produces probiotics and organic acids, improving palatability, but its efficiency is greatly affected by pretreatment, the conversion is incomplete, and the function is insufficient; through the synergy of enzymatic hydrolysis and fermentation, enzymatic hydrolysis provides precursor substances, and fermentation further transforms and modifies them to produce active substances, which meets the current industry development needs of green farming and efficient resource utilization.
[0004] Patent CN102948621B discloses a probiotic peptide biological feed additive, its preparation method and application. The above patent improves the gastrointestinal microenvironment of animals, has the dual biological effects of beneficial bacteria and active peptides, and increases the feed intake of animals.
[0005] The active peptides in the probiotic feed additives of the above-mentioned patents can stimulate animal immune function and improve animal immunity. They solve the problems of high bitter peptide content and poor palatability caused by acid, alkali and protease hydrolysis processes. However, there is still room for optimization in terms of enzymatic hydrolysis-fermentation synergy. This application solves the problem that a single additive cannot meet multiple needs such as nutrition and immunity at the same time.
[0006] Therefore, this application proposes an enzymatic hydrolysis-fermentation synergistic biotransformation process for biopeptide feed additives and its preparation method. Summary of the Invention
[0007] The purpose of this invention is to provide an enzymatic hydrolysis-fermentation synergistic biopeptide feed additive and its preparation method, so as to solve the technical problem mentioned in the background art that a single additive is difficult to meet multiple needs such as nutrition and immunity at the same time.
[0008] To achieve the above objectives, the present invention provides the following technical solution: an enzymatic hydrolysis-fermentation synergistic biopeptide feed additive, comprising fermented wet feed, wherein the fermented wet feed comprises, by weight: 100 parts rapeseed meal, 0.06-0.08 parts cellulase, 0.04-0.06 parts pectinase, 0.03-0.05 parts neutral protease, 1-3 parts compound microbial agent, 0.5-2 parts astragalus polysaccharide, 0.1-0.5 parts xylooligosaccharide, 1-3 parts linoleic acid, 100-110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid;
[0009] The compound microbial agent is a combination of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis.
[0010] Preferably, the method for preparing the fermented wet material includes the following steps:
[0011] The rapeseed meal was crushed using a pulverizer and sieved through a 40-60 mesh screen. The crushed and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added and stirred. Hydrochloric acid was added to adjust the pH value to 6-7 to obtain the mixture.
[0012] The mixed materials are placed in a fermenter and kept at 45-50℃ for 4-6 hours to obtain the enzymatically hydrolyzed material.
[0013] Cool the mixture to 30-37℃, add compound microbial agent to the enzymatically hydrolyzed material, stir, seal the fermentation tank, and ferment for 48-72 hours to obtain fermented wet material.
[0014] Preferably, the ratio of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis is 2:2:3.
[0015] Preferably, the nutritional indicators of the rapeseed meal are: crude protein content ≥36%, crude fiber ≤12%, glucosinolate content ≤25μmol / g, and moisture content ≤12%.
[0016] Preferably, the cellulase has an enzyme activity ≥1.5 × 10⁻⁶. 5 FPU / g, pectinase ≥2×10 5 U / g, neutral protease ≥1×10 5 U / g.
[0017] Preferably, the polysaccharide content of the Astragalus polysaccharide is ≥70%, and the purity of xylooligosaccharide is ≥95%.
[0018] Preferably, the method for preparing the traditional Chinese medicine decoction includes the following steps:
[0019] Mix houttuynia cordata, mugwort, licorice, vaccaria segetalis, tung oil, and amaranth, put them in a container, add 10 times the amount of distilled water, soak for 30 minutes, and then heat the soaked herbs to a boil for 30 minutes.
[0020] Use gauze to collect the first decoction. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter again using gauze.
[0021] The decoctions obtained from the two decoctions were mixed and then heated and concentrated to obtain a traditional Chinese medicine decoction containing 1g / ml of raw medicinal material.
[0022] Preferably, the mass ratio of Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata is 6:4:6:3:3:7.
[0023] Preferably, the method for preparing the feed additive includes the following steps:
[0024] S1. Dry the fermented wet material at 50℃ until the moisture content is ≤12%, crush the dried material with a pulverizer, and sieve it through a 40-60 mesh screen to obtain fermented dry powder.
[0025] S2. Place the sieved fermented dry powder and zeolite powder into a three-dimensional mixer and mix for 15-45 minutes to obtain the feed additive.
[0026] Preferably, the mass ratio of the fermented dry powder to the zeolite powder is 1:0.2.
[0027] Compared with the prior art, the beneficial effects of the present invention are:
[0028] 1. This invention prepares feed additives from components such as rapeseed meal, cellulase, neutral protease, and astragalus polysaccharide, realizing a biotransformation process of synergistic enzymatic hydrolysis and fermentation, establishing a process path of enzymatic hydrolysis followed by fermentation, organically combining the rapid and targeted degradation of exogenous enzymes with the deep fermentation and transformation of endogenous microorganisms, solving the problem that a single additive cannot simultaneously meet multiple needs such as nutrition and immunity, improving the nutritional value and utilization efficiency of feed raw materials, and enhancing the immunity and disease resistance of animals;
[0029] 2. This invention introduces a compound strain of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis to achieve a multi-strain spatiotemporal synergistic fermentation mode, which solves the problems of low fermentation efficiency and susceptibility to contamination by miscellaneous bacteria of single strains, improves the overall bioconversion efficiency of rapeseed meal substrate, and increases the abundance and diversity of active ingredients in the final product.
[0030] 3. By introducing traditional Chinese medicine decoction to replace conventional sterile water, this invention achieves the substitution of functional solvents and the direct implantation of active ingredients, solving the problem of ordinary fermented feed lacking specific antibacterial and anti-inflammatory functions, and improving the functional level and reliability of the product in disease prevention and animal health improvement.
[0031] 4. This invention achieves solidification and carrier fixation of active fermentation products by blending fermentation dry powder with zeolite powder, loading the fermentation powder onto zeolite powder particles with a porous structure, solving the problems of easy moisture absorption, clumping, and poor flowability of high-activity fermentation dry powder, and improving the physical stability and storage stability of the final product. Attached Figure Description
[0032] Figure 1 This is a schematic diagram of the fermentation wet material preparation process of the present invention;
[0033] Figure 2 This is a schematic diagram of the traditional Chinese medicine decoction preparation process of the present invention;
[0034] Figure 3 This is a schematic diagram of the feed additive preparation process of the present invention;
[0035] Figure 4 This is a schematic diagram of the compound microbial agent of the present invention. Detailed Implementation
[0036] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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.
[0037] Example 1, please refer to Figure 1 , Figure 2 , Figure 3 and Figure 4 A bioactive peptide feed additive based on enzymatic hydrolysis and fermentation includes fermented wet feed, which by weight comprises: 100 parts rapeseed meal, 0.08 parts cellulase, 0.06 parts pectinase, 0.05 parts neutral protease, 3 parts compound microbial agent, 2 parts astragalus polysaccharide, 0.5 parts xylooligosaccharide, 3 parts linoleic acid, 110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a ratio of 2:2:3. The feed additive preparation method includes the following steps:
[0038] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0039] Rapeseed meal was pulverized using a pulverizer and sieved through a 460-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0040] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0041] Furthermore, rapeseed meal, as a core substrate and protein source, provides the protein, carbohydrates, and a small amount of fat required for fermentation, which can be used to produce bioactive peptides. Rapeseed meal is rich in crude protein, making it a primary substrate for neutral proteases, which can enzymatically hydrolyze it into bioactive peptides. Simultaneously, rapeseed meal contains high levels of fiber, making it a target for cellulase and pectinase. Cellulase, in conjunction with pectinase, degrades the cellulose in rapeseed meal, disrupting plant cell wall structures and releasing encapsulated nutrients. It breaks down the cellulose and pectin in rapeseed meal into small-molecule sugars such as glucose and galacturonic acid, thus providing fermentable reducing sugars for subsequent fermentation with compound microbial agents, promoting the proliferation and metabolism of beneficial bacteria such as lactic acid bacteria and Bacillus. Neutral proteases can further degrade the large molecules in rapeseed meal... The protein is directionally hydrolyzed into small peptides and free amino acids. Hydrochloric acid is used to adjust the pH, ensuring the neutral protease is at its optimal pH, thus efficiently cleaving the peptide bonds. The peptides and amino acids produced by enzymatic hydrolysis serve as a high-quality nitrogen source for subsequent compound microbial fermentation, promoting rapid growth and acid production. Astragalus polysaccharide, an exogenous functional polysaccharide, acts as an immune enhancer, endowing the product with immunomodulatory functions. Introducing astragalus polysaccharide into feed additives can improve animal health and reduce disease incidence. Bioactive peptides produced from rapeseed meal enzymatic hydrolysis synergistically work with astragalus polysaccharide and xylooligosaccharides to enhance animal immune function and improve intestinal microecological balance. Linoleic acid, an unsaturated fatty acid, can regulate animal lipid metabolism and enhance the nutritional value of feed.
[0042] During the preparation of fermented wet feed, in the enzymatic hydrolysis stage, cellulase and pectinase act on rapeseed meal, breaking down the plant cell walls and releasing encapsulated proteins and starches, producing reducing sugars. Neutral proteases then act on the released proteins, hydrolyzing them into bioactive peptides and amino acids, creating a high-quality culture medium rich in small peptides, amino acids, and reducing sugars for subsequent fermentation. During fermentation, a compound inoculant containing *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bacillus subtilis* is inoculated. The reducing sugars and small peptides produced by enzymatic hydrolysis serve as high-quality carbon and nitrogen sources, which are rapidly utilized by probiotics, promoting fermentation. During its rapid growth and fermentation stage, *Lactobacillus plantarum* and *Lactobacillus acidophilus* produce acid, lowering the pH, inhibiting unwanted bacteria, and improving flavor. *Bacillus subtilis* consumes oxygen, creating an anaerobic environment. Xylooligosaccharides, as prebiotics, continuously promote the growth of probiotics. Linoleic acid, as a substrate for energy and functional fatty acids, is also present. During the growth and metabolism of probiotics, a variety of active substances are produced, which, together with bioactive peptides, astragalus polysaccharides, and xylooligosaccharides in the enzymatic hydrolysis products, form a synergistic effect, further enhancing the immunomodulatory function and intestinal microecological improvement effect of the feed additive, providing animals with more comprehensive nutritional support and health protection.
[0043] Example 2, please refer to Figure 1 , Figure 2 and Figure 3An enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive comprises fermented wet feed, which by weight includes: 100 parts rapeseed meal, 0.07 parts cellulase, 0.05 parts pectinase, 0.04 parts neutral protease, 3 parts compound microbial agent, 2 parts astragalus polysaccharide, 0.5 parts xylooligosaccharide, 3 parts linoleic acid, 110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a 2:2:3 ratio. The feed additive preparation method includes the following steps:
[0044] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0045] Rapeseed meal was pulverized using a pulverizer and sieved through a 60-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0046] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0047] Furthermore, a decoction of traditional Chinese medicine made from houttuynia cordata, artemisia argyi, licorice, vaccaria segetalis, potaspergillus, and amaranth is mixed with rapeseed meal, cellulase, and other materials to introduce multiple biological activities with antibacterial and anti-inflammatory properties into the feed additive. The decoction contains a large number of specific bioactive components released, transformed, and enriched during the fermentation and decoction process. During the enzymatic hydrolysis stage of preparing fermented wet feed, water-soluble active components in the decoction, such as polysaccharides, flavonoids, organic acids, and alkaloids, are fully dissolved and evenly dispersed throughout the rapeseed meal material system, serving as a direct source of active ingredients during the fermentation stage. The inoculated probiotics, including *Lactobacillus plantarum*, *Lactobacillus acidophilus*, and *Bacillus subtilis*, can utilize some of the sugars and amino acids in the decoction as nutrients for growth and participate in the microbial fermentation process. The antibacterial and anti-inflammatory components of the traditional Chinese medicine decoction, together with the bioactive peptides produced by enzymatic hydrolysis of rapeseed meal, the probiotics and their metabolites produced by fermentation, and functional components such as astragalus polysaccharides and xylooligosaccharides, constitute a multi-target, multi-level nutritional-microecological-immune-antibacterial complex functional system. This gives the feed additive good antibacterial and anti-inflammatory properties, improves the intestinal microecological balance of animals, increases feed utilization, enhances animal immunity, reduces disease incidence, and provides effective support for healthy livestock and poultry farming.
[0048] Example 3, please refer to Figure 1 , Figure 2 and Figure 3 A bioactive peptide feed additive based on enzymatic hydrolysis and fermentation includes fermented wet feed, which by weight comprises: 100 parts rapeseed meal, 0.08 parts cellulase, 0.06 parts pectinase, 0.05 parts neutral protease, 1 part compound microbial agent, 1 part astragalus polysaccharide, 0.4 parts xylooligosaccharide, 2 parts linoleic acid, 110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a 2:2:3 ratio. The feed additive preparation method includes the following steps:
[0049] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0050] Rapeseed meal was pulverized using a pulverizer and sieved through a 60-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0051] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0052] Furthermore, *Lactobacillus plantarum* and *Lactobacillus acidophilus* are used for rapid acid production. They utilize the reducing sugars and xylooligosaccharides produced by enzymatic hydrolysis for lactic acid fermentation, generating large amounts of lactic acid and acetic acid. This lowers the pH of the fermentation system, inhibits the growth of miscellaneous and putrefactive bacteria, and provides a stable acidic environment for the entire fermentation process. *Bacillus subtilis* is used for enzyme production and oxygen consumption. It can secrete abundant proteases, amylases, cellulases, etc. In the early stage of fermentation, *Bacillus subtilis* can rapidly consume oxygen in the material gaps, creating an anaerobic environment. At the same time, the proteases secreted by *Bacillus subtilis* can synergistically work with the neutral proteases added to the formula to further degrade proteins, generating more small peptides and amino acids. In the early stage of fermentation, *Bacillus subtilis* consumes oxygen, lowers the redox potential, and secretes enzymes to further degrade macromolecules. In the middle and late stages of fermentation, as oxygen is depleted and an anaerobic environment is formed, *Lactobacillus plantarum* and *Lactobacillus acidophilus* multiply in large quantities, dominating the fermentation process through acid production. Compared to single lactic acid bacteria, the combination of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis creates a sequential ecological niche in the initial aerobic environment, enabling relay fermentation. Rapeseed meal has a complex structure and contains anti-nutritional factors, limiting the processing capacity of single strains. In the compound inoculant, the enzyme production capacity of Bacillus subtilis can assist in the degradation of complex substrates such as cellulose and protein, thereby improving the overall bioconversion efficiency of this complex raw material, rapeseed meal. Bacillus subtilis creates an anaerobic environment, reducing the risk of growth inhibition of Lactobacillus plantarum due to early aerobic stress. The addition of Lactobacillus acidophilus increases the diversity of acid-producing bacteria, further enhancing the acid-producing and antibacterial ability, making the fermentation system more stable and less susceptible to contamination by other bacteria. The synergistic effect creates a better fermentation environment. Bacillus subtilis can colonize the animal intestine, inhibiting pathogens and forming a stronger probiotic community with Lactobacillus plantarum and Lactobacillus acidophilus in the intestine, synergistically improving the intestinal microecology.
[0053] Example 4, please refer to Figure 1 , Figure 2 and Figure 3 A bioactive peptide feed additive based on enzymatic hydrolysis and fermentation includes fermented wet feed, which by weight comprises: 100 parts rapeseed meal, 0.07 parts cellulase, 0.05 parts pectinase, 0.04 parts neutral protease, 2 parts compound microbial agent, 1 part astragalus polysaccharide, 0.3 parts xylooligosaccharide, 2 parts linoleic acid, 105 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a 2:2:3 ratio. The feed additive preparation method includes the following steps:
[0054] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0055] Rapeseed meal was pulverized using a pulverizer and sieved through a 60-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0056] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0057] Furthermore, blending zeolite powder with fermented dry powder allows the dry, bioactive fermented dry powder to be loaded onto the surface and interior of the porous structure of the zeolite powder. This effectively dilutes the highly active fermented dry powder, prevents clumping, and improves the physical flowability of the feed additive, making it easier to disperse evenly in subsequent feed processing and mixing. Zeolite powder also ensures product stability during storage, preventing mold or clumping due to moisture absorption. After ingestion, the porous structure of zeolite powder allows it to selectively adsorb harmful substances such as ammonium ions in the animal's intestines through ion exchange, effectively adsorbing harmful metabolites such as ammonia and hydrogen sulfide. It is fixed and then excreted with feces, thereby reducing the concentration of ammonia in the intestines and improving intestinal health. At the same time, the addition of zeolite powder can enhance the slow-release effect of bioactive components in feed additives, prolong their action time in the animal's intestines, and further improve the utilization rate of nutrients. Zeolite powder is rich in various mineral elements such as calcium, magnesium, and potassium, which can serve as a source of trace elements required for animal growth, help regulate electrolyte balance, and promote metabolism and growth. In addition, zeolite powder is chemically inert and does not easily react with other active additives in feed, effectively maintaining the original functions of each component and ensuring stable product performance.
[0058] Example 5, please refer to Figure 1 , Figure 2 , Figure 3 and Figure 4 A bioactive peptide feed additive based on enzymatic hydrolysis and fermentation includes fermented wet feed, which by weight comprises: 100 parts rapeseed meal, 0.06 parts cellulase, 0.04 parts pectinase, 0.03 parts neutral protease, 1 part compound microbial agent, 0.5 parts astragalus polysaccharide, 0.1 parts xylooligosaccharide, 1 part linoleic acid, 100 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a 2:2:3 ratio. The feed additive preparation method includes the following steps:
[0059] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0060] Rapeseed meal was pulverized using a pulverizer and sieved through a 60-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 337°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0061] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0062] Comparative Example 1: An enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive, comprising fermented wet feed, wherein the fermented wet feed comprises, by weight: 100 parts rapeseed meal, 0.08 parts cellulase, 0.06 parts pectinase, 0.05 parts neutral protease, 3 parts *Lactobacillus plantarum*, 2 parts astragalus polysaccharide, 0.5 parts xylooligosaccharide, 3 parts linoleic acid, 110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid; the feed additive preparation method includes the following steps:
[0063] Mix Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata in a mass ratio of 6:4:6:3:3:7 and place them in a container. Add 10 times the amount of distilled water and soak for 30 minutes. Heat the soaked herbs to a boil and continue for 30 minutes. Collect the first decoction with gauze. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter with gauze again. Mix the decoctions obtained from the two decoctions and heat to concentrate to obtain a Chinese herbal decoction with a raw herb content of 1g / ml.
[0064] Rapeseed meal was pulverized using a pulverizer and sieved through a 460-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0065] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0066] Comparative Example 2: An enzymatic hydrolysis-fermentation synergistic biopeptide feed additive, comprising fermented wet feed, which by weight includes: 100 parts rapeseed meal, 0.08 parts cellulase, 0.06 parts pectinase, 0.05 parts neutral protease, 3 parts compound microbial agent, 2 parts astragalus polysaccharide, 0.5 parts xylooligosaccharide, 3 parts linoleic acid, 110 parts sterile water, and an appropriate amount of hydrochloric acid. The compound microbial agent contains Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis in a 2:2:3 ratio. The feed additive preparation method includes the following steps:
[0067] Rapeseed meal was pulverized using a pulverizer and sieved through a 460-mesh sieve. The pulverized and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added, and the mixture was stirred. Hydrochloric acid was added to adjust the pH to 7, resulting in a mixture. The mixture was placed in a fermentation tank and kept at 50°C for 6 hours to obtain enzymatically hydrolyzed material. The temperature was then lowered to 37°C, and a compound microbial agent was added to the enzymatically hydrolyzed material. The mixture was stirred, the fermentation tank was sealed, and fermentation was carried out for 72 hours to obtain fermented wet material.
[0068] The fermented wet material was dried at 50℃ until the moisture content was ≤12%. The dried material was then crushed using a pulverizer and sieved through a 60-mesh sieve to obtain fermented dry powder. The sieved fermented dry powder and zeolite powder were then placed in a three-dimensional mixer at a mass ratio of 1:0.2 and mixed for 45 minutes to obtain a feed additive.
[0069] Performance testing
[0070] Test 1: Growth performance test. 28-day-old weaned piglets with similar weights were selected and fed a basal diet without feed additives and a basal diet with feed additives produced in Examples 1-5 and Comparative Examples 1-2, respectively. The amount of feed additives added was 0.1%. After 6 weeks of feeding, the average daily weight gain and feed conversion ratio were calculated.
[0071] Test 2: Serum index testing of animals. Animals were fed a basal diet without feed additives and a basal diet with feed additives produced in Examples 1-5 and Comparative Examples 1-2. After feeding, blood was collected. The collected blood was tilted and allowed to stand at room temperature for about 30 minutes to coagulate naturally. After centrifugation at 4°C and 3000 rpm for 10 minutes, the clear light yellow liquid in the upper layer was collected. Immunoglobulin levels and inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA).
[0072] The performance test results are shown in Table 1 below:
[0073] Table 1. Performance Test Results
[0074] In summary, based on the performance test results of Examples 1-5 and Comparative Examples 1-2, the compound microbial agent is superior to feed additives with only a single microbial species in terms of growth and immune indicators compared to single-species microbial agents, and the use of traditional Chinese medicine decoction can improve the anti-inflammatory effect.
[0075] Working principle: Before fermentation, rapeseed meal is pretreated with cellulase, pectinase and neutral protease. Cellulase and pectinase work together to degrade the cell walls of rapeseed meal, releasing large molecular proteins, starch and other nutrients that are wrapped inside the cells, producing fermentable reducing sugars, which increases the substrate surface area that subsequent enzymes and microorganisms can contact. Neutral protease directionally hydrolyzes large molecular proteins into small molecular bioactive peptides and free amino acids.
[0076] After enzymatic hydrolysis, a compound probiotic agent consisting of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis is introduced for fermentation. Lactobacillus plantarum and Lactobacillus acidophilus produce acid to inhibit the growth of other bacteria, while Bacillus subtilis provides oxygen to create an anaerobic environment.
[0077] By mixing zeolite powder with fermentation dry powder, the porous structure and ion exchange capacity of zeolite powder are utilized to act as a carrier, adsorb moisture and harmful gases, improve product flowability, and continuously adsorb harmful substances such as ammonia in the animal intestines to improve the internal environment.
[0078] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.
Claims
1. A bioactive peptide feed additive with enzymatic hydrolysis-fermentation synergistic effect, characterized in that: The mixture includes fermented wet material, which by weight comprises: 100 parts rapeseed meal, 0.06-0.08 parts cellulase, 0.04-0.06 parts pectinase, 0.03-0.05 parts neutral protease, 1-3 parts compound microbial agent, 0.5-2 parts astragalus polysaccharide, 0.1-0.5 parts xylooligosaccharide, 1-3 parts linoleic acid, 100-110 parts traditional Chinese medicine decoction, and an appropriate amount of hydrochloric acid; The compound microbial agent is a combination of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis.
2. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The method for preparing the fermented wet material includes the following steps: The rapeseed meal was crushed using a pulverizer and sieved through a 40-60 mesh screen. The crushed and sieved rapeseed meal was then mixed with cellulase, pectinase, neutral protease, astragalus polysaccharide, xylooligosaccharide, and linoleic acid. Traditional Chinese medicine decoction was added and stirred. Hydrochloric acid was added to adjust the pH value to 6-7 to obtain the mixture. The mixed materials are placed in a fermenter and kept at 45-50℃ for 4-6 hours to obtain the enzymatically hydrolyzed material. Cool the mixture to 30-37℃, add compound microbial agent to the enzymatically hydrolyzed material, stir, seal the fermentation tank, and ferment for 48-72 hours to obtain fermented wet material.
3. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The ratio of Lactobacillus plantarum, Lactobacillus acidophilus, and Bacillus subtilis is 2:2:
3.
4. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The nutritional indicators of the rapeseed meal are as follows: crude protein content ≥36%, crude fiber ≤12%, glucosinolate content ≤25μmol / g, and moisture ≤12%.
5. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The cellulase activity is ≥1.5×10⁻⁶. 5 FPU / g, pectinase ≥2×10 5 U / g, neutral protease ≥1×10 5 U / g.
6. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The polysaccharide content of the Astragalus polysaccharide is ≥70%, and the purity of xylooligosaccharide is ≥95%.
7. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 1, characterized in that: The preparation method of the traditional Chinese medicine decoction includes the following steps: Mix houttuynia cordata, mugwort, licorice, vaccaria segetalis, tung oil, and amaranth, put them in a container, add 10 times the amount of distilled water, soak for 30 minutes, and then heat the soaked herbs to a boil for 30 minutes. Use gauze to collect the first decoction. Add 8 times the amount of distilled water to the filtered residue and repeat the above decoction process. Filter again using gauze. The decoctions obtained from the two decoctions were mixed and then heated and concentrated to obtain a traditional Chinese medicine decoction containing 1g / ml of raw medicinal material.
8. The enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 7, characterized in that: The mass ratio of Houttuynia cordata, Artemisia argyi, Glycyrrhiza uralensis, Vaccaria segetalis, Potentilla chinensis, and Amaranthus trifoliata is 6:4:6:3:3:
7.
9. A method for preparing an enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive, applicable to the enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to any one of claims 1-8, characterized in that: The method for preparing the feed additive includes the following steps: S1. Dry the fermented wet material at 50℃ until the moisture content is ≤12%, crush the dried material with a pulverizer, and sieve it through a 40-60 mesh screen to obtain fermented dry powder. S2. Place the sieved fermented dry powder and zeolite powder into a three-dimensional mixer and mix for 15-45 minutes to obtain the feed additive.
10. The method for preparing an enzymatic hydrolysis-fermentation synergistic bioactive peptide feed additive according to claim 9, characterized in that: The mass ratio of the fermented dry powder to the zeolite powder is 1:0.2.