Complex enzyme preparation applied to broiler feed and preparation method of complex enzyme preparation

Abstract

The invention discloses a compound enzyme preparation applied to broiler feed and a preparation method of the compound enzyme preparation. The enzyme preparation is prepared from the following steps from inside to outside: 1) taking neutral compound enzyme as a core, and carrying out primary embedding by adopting a sporopollen outer-wall capsule to provide protection and mucous membrane adhesiveness; 2) the composition is composed of bile acid analogues, short-chain fatty acid, astragalus polysaccharide and hydroxypropyl methyl cellulose, and aims to build a favorable intestinal environment in advance and assist in activating enzyme activity; 3) the enzyme protection response layer contains konjac mannose oligomer, betaine (betaine) and methacrylic acid copolymer, and is used for chelating metal ions and realizing pH responsive release of intestinal tracts; 4, the external gel protection shell is prepared by mixing lac and sodium alginate, and the granules are endowed with good mechanical strength and completeness. Enzyme activity is effectively protected through multi-layer collaborative design, targeted and efficient release and action of enzyme in intestinal tracts are achieved, the digestion and utilization rate of broiler chicken to feed nutrition is remarkably increased, and emission of undigested substances is reduced.

Description

Technical Field

[0001] This invention belongs to the field of feed additive technology, specifically relating to a compound enzyme preparation for use in broiler feed and its preparation method. Background Technology

[0002] The broiler industry faces the dual pressures of improving production efficiency and reducing its environmental footprint. Approximately 20-30% of the nutrients in feed are not fully digested and absorbed by animals. These undigested nutrients (such as protein and phosphorus) are excreted in feces, not only wasting resources but also generating large amounts of greenhouse gases and nitrogen and phosphorus pollutants through microbial action, putting pressure on the environment. Enzyme preparations, as highly efficient feed additives, can effectively degrade large, indigestible nutrients in feed (such as starch, protein, cellulose, and phytic acid phosphorus), improving nutrient absorption efficiency, reducing the excretion of undigested substances, and thus reducing carbon emissions during the farming process. This is one of the key technologies for achieving green farming.

[0003] Currently, commonly used enzyme preparations in feed include phytase, protease, and non-starch polysaccharide enzymes (such as xylanase and β-glucanase). However, existing enzyme preparations for broiler feed suffer from several technical challenges in practical applications, severely limiting their functionality: First, the enzyme preparations themselves have poor stability and are prone to conformational changes during high-temperature and high-pressure feed processing, leading to loss of activity. Second, they lack targeting; after entering the broiler's stomach, they are easily destroyed by gastric acid and pepsin, and metal ions in the stomach environment inhibit enzyme activity, causing a large number of enzymes to become inactive before reaching the intestines, resulting in extremely low bioavailability. Third, existing enzyme preparations are mostly single-function components, failing to form a complete "protection-activation-synergy" system. Even when some products use encapsulation technology, they are mostly simple single- or double-layer encapsulations, unable to achieve precise slow release and long-term maintenance of enzyme activity, and do not take into account the improvement of intestinal microecology and the maintenance of the body, making it difficult to fully exert the catalytic efficacy of the enzyme preparations. Summary of the Invention

[0004] The technical problem to be solved: This invention addresses the pain points of enzyme preparations being easily inactivated in feed processing and the gastric environment, and having poor intestinal targeting, through a multi-layer encapsulation structure. Multiple functional components that promote digestion and improve the intestines work synergistically to achieve targeted and sustained release of enzymes in the intestines and long-term maintenance of their activity. At the same time, it improves the digestibility of broiler feed, promotes broiler growth, and reduces carbon emissions from farming. It is practical, environmentally friendly, and functional. The preparation process is simple and controllable, making it suitable for large-scale industrial production.

[0005] Technical solution: A method for preparing a compound enzyme preparation for broiler feed, comprising the following steps: S1. Take pollen, crush and sieve it to obtain pollen powder, add acetone, gently stir and reflux at 50℃ for 6 hours, vacuum filter to collect defatted spores, air dry, mix with phosphoric acid solution, stir and reflux at constant temperature, then wash, finally vacuum filter to collect, and dry to obtain spore-coated capsule powder. S2. The outer wall capsule powder of spore-spearin was reconstituted with water to prepare a uniform spore-spearin suspension. The enzyme suspension was slowly added to the spore-spearin suspension, stirred in a constant temperature water bath, and then freeze-dried under vacuum to obtain the core enzyme spore-spearin outer wall capsules. S3. Bile acid analogues, short-chain fatty acid mixtures, and astragalus polysaccharides were dissolved in water and stirred. The core enzyme, sporophytin, was added to the outer wall capsules, emulsified and sheared, and finally hydroxypropyl methylcellulose was added and emulsified and sheared again. Then, the mixture was freeze-dried to obtain a double-layer embedded lyophilized powder. S4. Konjac oligomannose, betaine and betaine phosphate, methacrylic acid copolymer, water were added and stirred to make a mixture, double-layer embedded freeze-dried powder was added to the mixture, homogenized and spray-dried to obtain a preliminary enzyme preparation; S5. Add water to hawthorn powder, stir and dissolve at a constant temperature, filter to obtain a clear hawthorn powder solution, slowly pour sodium alginate solution into the clear hawthorn solution while stirring, then add glycerol and preliminary enzyme preparation and continue stirring, shear emulsify to obtain an emulsion, slowly drip the emulsion into cold calcium chloride solution, let stand, separate the gel particles, wash with water and dry, and sieve to obtain a compound enzyme preparation.

[0006] Furthermore, in step S1, the mass ratio of pollen powder to acetone is 1:(3-5); the temperature of gentle stirring and reflux is 45-55℃ for 5-8 hours; the air drying time is 12-16 hours; the concentration of phosphoric acid solution is 70-85 wt.%; and the temperature of constant temperature stirring and reflux treatment is 65-70℃ for 25-30 hours.

[0007] Furthermore, the washing method in step S1 is as follows: after washing with hot water 3-5 times, wash with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide in sequence, then wash with hot water 3-5 times, then wash with hot acetone and hot ethanol in sequence, and finally wash with hot water 3-5 times.

[0008] Furthermore, in step S2, the mass ratio of the spore-forming outer wall capsule powder to water is 1:(9-10); the mass ratio of the enzyme suspension to the spore-forming suspension is 1:(2-4); the temperature of the constant temperature water bath stirring is 20-25℃, the rotation speed is 150-250rpm, and the time is 30-50min.

[0009] Furthermore, in step S2, the enzyme suspension is composed of enzymes with the following enzyme activity units: neutral protease 50,000-200,000 U / g, phytase 5,000-20,000 U / g, xylanase 10,000-50,000 U / g, β-glucanase 5,000-30,000 U / g, amylase 5,000-25,000 U / g; and lipase 800-1,000 U / g.

[0010] Furthermore, in step S3, the bile acid analogue is taurocholic acid; the short-chain fatty acid mixture is a mixture of acetic acid, propionic acid, and butyric acid in a mass ratio of (1-2):(1-2):(1-2); the mass ratio of bile acid analogue, short-chain fatty acid mixture, astragalus polysaccharide, hydroxypropyl methylcellulose, and core enzyme spore powder outer wall capsules is (2-3):(2-3):(1-2):(1-2):(12-15); the material-to-liquid ratio of water is 1:(4-8); the stirring and dissolving temperature is 25-30℃, and the stirring speed is 200-300 rpm; the emulsification and shearing speed is 2000-3000 rpm, and the time is 10-20 min.

[0011] Furthermore, in step S4, the methacrylic acid copolymer is L-type; the mass ratio of konjac oligomannose, betaine, betaine phosphate, methacrylic acid copolymer and water is (2-3):(1-2):(1-2):(2-3):(50-80); the ratio of double-layer embedded freeze-dried powder to mixed liquid is 1:(5-8); and the homogenization pressure is 10-15 MPa.

[0012] Furthermore, in step S5, the mass ratio of hawthorn powder to water is 1:(15-25); the constant temperature stirring dissolution temperature is 50-55℃, the rotation speed is 300-500rpm, and the time is 20-30min; the concentration of sodium alginate solution is 3-5wt.%; the mass ratio of sodium alginate solution, hawthorn clarified solution, glycerol, and preliminary enzyme preparation is (10-15):(20-40):1:(4-10); the stirring speed is 100-200rpm, and the time is 20-30min; the shear emulsification speed is 2000-4000rpm, and the time is 10-15min; the concentration of cold calcium chloride solution is 2-3wt.%, and the temperature is 4-5℃; the standing time is 10-15min; and the number of water washings is not less than 3.

[0013] The compound enzyme preparation prepared by the above method is used in broiler feed.

[0014] The present invention also includes the application of the above-mentioned compound enzyme preparations used in broiler feed in feeds that improve digestibility and reduce carbon emissions from broiler farming.

[0015] Beneficial effects: 1. The sporopolleninexine capsules of this invention have a natural rigid outer wall structure. Their molecular structure contains a large amount of cellulose, lignin, and sporopollenine polymers, forming a dense, porous protective barrier. These sporopolleninexine capsules (SECs) have advantages such as uniform structural morphology, large internal cavity, good elasticity, resistance to physicochemical processes, and antioxidant activity. Therefore, SECs can resist changes in pH, ionic strength, temperature, light, oxygen, and mechanical stress. Simultaneously, the sporopollenine surface has good adhesion, allowing it to tightly bond with the second layer of methacrylic acid copolymer, providing a stable substrate for subsequent mixed layer encapsulation. Compared to traditional encapsulation materials, this method leaves no residue and provides more durable and safer protection.

[0016] 2. The bile acid analogue and short-chain fatty acid of this invention synergistically activate intestinal enzyme activity and provide a suitable intestinal environment. Astragalus polysaccharide is responsible for immune regulation and enzyme protection. The three components form a polymer with methacrylic acid copolymer, which can achieve four-fold synergy of shaping, activation, enzyme protection and immunity, with more comprehensive functions and better shapeability.

[0017] 3. The first layer of this invention, sporophytin, is responsible for basic physical protection and adhesion to the substrate. The second layer, a mixture of components, is responsible for activation, creating a suitable environment and immunity for the enzyme. The third layer is responsible for strengthening enzyme protection, removing enzyme inhibitors, and synergizing with prebiotics. Finally, the gel encapsulation is more tight, allowing free substances that cannot form a three-layer encapsulation structure to be encapsulated within the gel. This provides a safety net to prevent leakage during the initial encapsulation process, serving as the final guarantee. Each layer has a clear division of labor and works synergistically, solving the problems of incomplete encapsulation protection, inaccurate release, easy detachment of the encapsulation, and single function in traditional methods. This ensures the maximum utilization of enzyme activity and the functions of each layer's components.

[0018] 4. The inactivated metabiotics of *Lactobacillus fermentum* used in this invention can adhere to the intestinal mucosa, forming a favorable microecological environment, while strengthening the brush border and tight junctions of the intestinal epithelium, rapidly improving gastrointestinal motility and flatulence; *Bifidobacterium longum* ES1, derived from breast milk, is more easily accepted by consumers, and retains its cell wall, metabolites, and other active ingredients after inactivation, thus inhibiting pathogenic bacteria and alleviating digestive symptoms; *Lactobacillus paracasei* DG, after inactivation, can significantly increase postprandial blood amino acid concentration, indicating that it can improve intestinal absorption of nutrients; the three bacteria promote digestion and regulate gastrointestinal health through different pathways, and these three metabiotics focus on three core links: physical barrier, immune regulation, and nutrient absorption, forming a synergistic defense network.

[0019] 5. This invention utilizes a core enzyme to efficiently degrade feed nutrients, improving feed digestibility and broiler daily weight gain while reducing the feed conversion ratio. After being absorbed by the intestines, Astragalus polysaccharides stimulate the proliferation of intestinal mucosal immune cells, enhance intestinal barrier function, reduce the incidence of intestinal diseases in broilers, lower mortality, and simultaneously increase serum immunoglobulin levels, achieving synergistic improvement in broiler growth and immunity. The improved feed digestibility reduces the excretion of undigested nutrients, lowering the emission of greenhouse gases such as methane and carbon dioxide in broiler feces, making it green, safe, and free of residue risks.

[0020] 6. This invention breaks through the functional limitations of traditional enzyme preparations that rely solely on catalysis. By incorporating natural digestive aids, it not only utilizes the gelling properties of hawthorn but also synergistically enhances feed digestibility and promotes broiler growth with the core enzymes. Furthermore, it incorporates probiotic components such as astragalus polysaccharides and konjac oligomannose to promote the proliferation of beneficial intestinal bacteria, inhibit the growth of harmful bacteria, and improve the intestinal microecological structure. Simultaneously, astragalus polysaccharides enhance intestinal barrier function, alleviate intestinal inflammation, reduce the occurrence of intestinal diseases, and improve the safety of broiler products.

[0021] 7. This invention utilizes the synergistic effect of multiple enzymes: neutral protease breaks down complex proteins in plant-based protein raw materials, releasing small peptides and amino acids, improving protein digestibility, and reducing nitrogen emissions in feces at the source; phytase breaks down phytic acid phosphorus, releasing bound phosphorus, significantly improving phosphorus utilization, reducing phosphorus emissions in feces, and simultaneously releasing proteins and trace elements bound to phytic acid; xylanase and β-glucanase synergistically disrupt grain cell walls, breaking down indigestible non-starch polysaccharides, reducing chyme viscosity, and releasing bound starch and protein; in addition, amylase and lipase assist in the breakdown of starch and fat, achieving simultaneous and efficient utilization of the three core nutrients: protein, energy, and phosphorus.

[0022] 8. All encapsulation materials in this invention are of natural origin or food-grade products, with good biocompatibility and biodegradability. After entering the broiler's body, they can be slowly degraded without residue or side effects. The hawthorn in the outermost gel protective shell not only plays a protective role in gel encapsulation, but also contains organic acids that can play an auxiliary role in digestion, promote gastrointestinal motility, increase gastric juice secretion, and work synergistically with core enzymes, short-chain fatty acids and other components to further optimize the digestive environment and improve overall digestive efficiency. Detailed Implementation

[0023] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments: Example 1

[0024] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 2

[0025] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 25h. After washing with hot water 5 times, the mixture was washed with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide in sequence. After washing with hot water 5 times, the mixture was washed with hot acetone and hot ethanol in sequence. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 3

[0026] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 25g of enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 4

[0027] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 50000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 5000U / g, amylase 25000U / g, and lipase 800U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 5

[0028] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 15g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 6

[0029] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 20g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Example 7

[0030] A method for preparing a compound enzyme preparation for use in broiler feed includes the following steps: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 6g of taurocholic acid, 6g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 4g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 20g of the core enzyme spore-forming outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 4g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 20g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Comparative Example 1

[0031] The difference between this comparative example and Example 1 is that it does not contain sporoquinone encapsulation, but is directly mixed with enzymes, as detailed below: S1. The enzyme suspension is composed of enzymes with the following enzyme activity units: neutral protease 200,000 U / g, phytase 20,000 U / g, xylanase 50,000 U / g, β-glucanase 30,000 U / g, amylase 25,000 U / g, and lipase 1,000 U / g. S2. Preparation of encapsulated lyophilized powder: 6g of taurocholic acid, 6g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 4g of astragalus polysaccharide were mixed and 88mL of water were added. The mixture was stirred and dissolved at 30℃ and 300rpm. 20g of the core enzyme spore-forming capsule outer wall was added and emulsified and sheared at 3000rpm for 10min. Finally, 4g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain the encapsulated lyophilized powder. S3. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of encapsulated lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S4. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Comparative Example 2

[0032] The difference between this comparative example and Example 1 is that bile acid analogs, short-chain fatty acid mixtures, astragalus polysaccharides, and hydroxypropyl methylcellulose are not added, as detailed below: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 18g of the core enzyme sporophytin outer wall capsules were added to the mixture. After homogenization at 15MPa, the mixture was spray-dried to obtain the preliminary enzyme preparation. S4. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Comparative Example 3

[0033] The difference between this comparative example and Example 1 is that it lacks the outer hawthorn-sodium alginate gel, as detailed below: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, homogenized at 15MPa, and then spray-dried to obtain a compound enzyme preparation. Comparative Example 4

[0034] The difference between this comparative example and Example 1 is that no bile acid analogues or short-chain fatty acid mixtures are added, as detailed below: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 70℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer encapsulated lyophilized powder: 2g of Astragalus polysaccharide was mixed with 88mL of water and stirred at 30℃ and 300rpm to dissolve. 12g of the core enzyme spore-forming outer wall capsule was added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. Then, the mixture was freeze-dried to obtain double-layer encapsulated lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 10g of hawthorn powder to 200g of water and stir at 55℃ and 400rpm for 25min to dissolve. Filter to obtain a clear hawthorn powder solution. Slowly pour 10g of 5wt.% sodium alginate solution into 40g of hawthorn clear solution while stirring. Then add 1g of glycerol and 10g of preliminary enzyme preparation. Continue stirring at 150rpm for 25min. Shear emulsify at 3000rpm for 12min to obtain an emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃. Let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation. Comparative Example 5

[0035] The difference between this comparative example and Example 1 is that hawthorn powder is not added, as detailed below: S1. Preparation of spore-forming outer wall capsule powder: 100g of rapeseed pollen was pulverized and passed through a 100-mesh sieve to obtain pollen powder. 300g of acetone was added, and the mixture was gently stirred and refluxed at 50℃ for 6h. The defatted spores were collected by vacuum filtration and air-dried for 14h. The mixture was then mixed with an 85wt.% phosphate solution and stirred and refluxed at 7℃ for 30h. After washing with hot water 5 times, the mixture was washed successively with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide. After washing with hot water 5 times, the mixture was washed successively with hot acetone and hot ethanol. Finally, the mixture was washed with hot water 5 times, collected by vacuum filtration, and dried to obtain spore-forming outer wall capsule powder. S2. Preparation of core enzyme spore-spraying capsules: 10g of spore-spraying capsule powder was dissolved in 90g of water to prepare a uniform spore-spraying suspension. 50g of the enzyme suspension was slowly added to the spore-spraying suspension. The mixture was stirred in a constant temperature water bath at 200rpm for 40min at 25℃, and then freeze-dried under vacuum to obtain core enzyme spore-spraying capsules. The enzyme suspension was composed of enzymes with the following enzyme activity units: neutral protease 200000U / g, phytase 20000U / g, xylanase 50000U / g, β-glucanase 30000U / g, amylase 25000U / g, and lipase 1000U / g. S3. Preparation of double-layer embedded lyophilized powder: 3g of taurocholic acid, 3g of a mixture of acetic acid, propionic acid and butyric acid (mass ratio 1:1:1) and 2g of astragalus polysaccharide were mixed and 88mL of water was added. The mixture was stirred and dissolved at 30℃ and 300rpm. 12g of the core enzyme sporophytin outer wall capsules were added and emulsified and sheared at 3000rpm for 10min. Finally, 2g of hydroxypropyl methylcellulose was added and emulsified and sheared again for 10min. The mixture was then freeze-dried to obtain double-layer embedded lyophilized powder. S4. Preparation of preliminary enzyme preparation: 6g of konjac oligomannose, 4g of betaine, 4g of betaine phosphate, and 6g of L-type methacrylic acid copolymer were mixed with 150g of water to form a mixture. 35g of double-layer embedded lyophilized powder was added to the mixture, and after homogenization at 15MPa, it was spray-dried to obtain the preliminary enzyme preparation. S5. Preparation of compound enzyme preparation: Add 1g of glycerol and 10g of preliminary enzyme preparation to 50g of 5wt.% sodium alginate solution, stir at 150rpm for 25min, and shear emulsify at 3000rpm for 12min to obtain emulsion. Slowly drop the emulsion into a 3wt.% cold calcium chloride solution at 4℃, let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain compound enzyme preparation. Comparative Example 6

[0036] The difference between this comparative example and Example 1 is that the enzyme suspension is directly mixed with maltodextrin and conventionally spray-dried, as follows: S1. The enzyme suspension is composed of enzymes with the following enzyme activity units: neutral protease 200,000 U / g, phytase 20,000 U / g, xylanase 50,000 U / g, β-glucanase 30,000 U / g, amylase 25,000 U / g, and lipase 1,000 U / g. S2. Preparation of compound enzyme preparation: 50g enzyme suspension and 50g maltodextrin are mixed and spray-dried to obtain compound enzyme preparation. Comparative Example 7

[0037] The difference between this comparative example and Example 1 is that the enzyme suspension is directly mixed with sodium alginate solution to form gel beads, as detailed below: S1. The enzyme suspension is composed of enzymes with the following enzyme activity units: neutral protease 200,000 U / g, phytase 20,000 U / g, xylanase 50,000 U / g, β-glucanase 30,000 U / g, amylase 25,000 U / g, and lipase 1,000 U / g. S3. Mix 50g of 5wt.% sodium alginate solution with 50g of enzyme suspension, continue stirring at 150rpm for 25min, then slowly drop it into a 3wt.% cold calcium chloride solution at 4℃, let stand for 15min, separate the gel particles, wash with water 3 times, dry, and sieve to obtain the compound enzyme preparation.

[0038] Performance testing: 1. Enzyme activity retention rate determination experiment Preparation of artificial gastric fluid: 1.0 g of pepsin (1:10000 activity), pH adjusted to 3.0 with concentrated hydrochloric acid, 2.0 g of sodium chloride, and water added to 1000 mL. Add 50 mL of the sample to the artificial gastric fluid, incubate at 37°C with shaking (100 rpm) for 2 h, immediately remove, cool to 4°C in an ice-water bath to terminate the reaction, centrifuge at 4000 rpm for 10 min, collect the precipitate, wash twice with pH 7.0 phosphate buffer, determine the total enzyme activity before and after gastric digestion, and calculate the enzyme activity retention rate.

[0039] Retention rate = (Total enzyme activity after gastric juice treatment / Initial total enzyme activity) × 100% The results are shown in Table 1. The compound enzyme preparations in the examples maintained high enzyme activity after gastric digestion, allowing them to function more efficiently in the intestines. The comparative examples showed lower retention rates, especially in Comparative Examples 6 and 7, where the enzyme retention rate was extremely low under the traditional process, making it difficult for them to exert their effects. Comparative Example 1, lacking the protection of the spore-forming outer capsule, also showed a significantly reduced enzyme activity retention rate.

[0040] Table 1. Retention rate of compound enzyme preparations after gastric digestion.

[0041] 2. Stability evaluation indicators Accelerated storage experiments were initiated at 30℃±2℃, with the enzymes sealed in aluminum foil bags and stored in the dark for 5 days. Initial and final enzyme activities were measured, and the enzyme activity loss rate was calculated.

[0042] Enzyme activity loss rate = (Initial enzyme activity - Enzyme activity after storage) / Initial enzyme activity × 100% The results are shown in Table 2. The loss rate of all examples was less than 17%, which means that they can have a longer shelf life under normal temperature or refrigeration conditions. Example 7 was the best, possibly because the double-layer encapsulated lyophilized powder components were doubled, and HPMC formed a denser matrix, blocking oxygen and moisture. The loss rates of comparative examples 1-5, which lacked relevant components or protective layers, were higher than those of the examples. Examples 6 and 7 suffered severe losses and were difficult to use as effective formulations.

[0043] Table 2 Loss rate of compound enzyme preparation after storage experiment

[0044] 3. Application effect verification One-day-old AA broilers were selected. The control group was fed a basal diet, while the experimental group was fed a basal diet plus 150 g / ton of the compound enzyme preparation prepared in the examples or comparative examples. The initial and final weights of the broilers were recorded, and the feed conversion ratio (F / G) was measured. Feed intake was recorded after the start of the experiment. Fecal and urine mixtures of chickens in each group were collected, air-dried, and their protein and other nutrient contents, phosphorus and nitrogen contents were measured. The apparent digestibility of nutrients was measured, and the average value and the reduction rate of phosphorus (nitrogen) content in feces and urine were calculated.

[0045]

[0046] The feed conversion ratio (FCR) is a core indicator for measuring the efficiency of broiler farming. It is defined as the amount of feed consumed per unit of body weight gain in broilers. A lower FCR indicates higher feed utilization, lower farming costs, and more significant farming benefits. Based on the experimental data in Table 3, the FCRs of each group showed significant differences. Example 1 had a FCR of 1.68 ± 0.06, a significant decrease of 9.19% compared to the control group. Comparative Example 6 was closest to the control group, while Comparative Example 5 had the lowest FCR, but it was still higher than Example 1. It is speculated that although this comparative example retained the core enzyme and some encapsulation structure, it lacked the hawthorn component or adjusted the four-layer encapsulation ratio, resulting in insufficient digestive aid and incomplete synergistic effect. These results fully demonstrate that the enzyme preparation in this technical solution can effectively improve feed utilization and reduce feed waste.

[0047] Dry matter digestibility and protein digestibility are important indicators for measuring the gastrointestinal digestive function of broilers. Higher digestibility indicates more efficient utilization of feed nutrients and less undigested nutrients. In Table 3, the digestibility of Example 1 was significantly higher than that of the control group and the comparative group, with a clear difference. Example 1 had the highest protein digestibility, corresponding to the lowest feed conversion ratio (FCR), and the best broiler weight gain, confirming the principle that "sufficient protein digestion and absorption can effectively promote broiler growth and reduce the FCR." Simultaneously, the improved protein digestibility reduced the emission of crude protein in feces and decreased the release of ammonia gas during fecal degradation. Combined with the improved dry matter digestibility, this further strengthened the "low-carbon and environmentally friendly" advantages of this formulation, achieving a dual improvement in both breeding and environmental benefits.

[0048] Table 3 Feed conversion ratio and digestibility of broilers fed with compound enzyme preparations

[0049] The reduction rate of phosphorus (nitrogen) content in feces and urine is a key environmental indicator for measuring the efficiency of phosphorus utilization in feed and reducing pollution. A higher reduction rate indicates a higher proportion of nitrogen and phosphorus in the feed being digested and absorbed by broilers, resulting in less nitrogen and phosphorus emissions in the feces. This reduces phosphorus resource waste and the environmental pollution risk from fecal emissions. Table 4 shows that Example 1 showed a significant reduction in phosphorus (nitrogen) content in feces and urine compared to the control group. While the comparative examples also showed a reduction compared to the control group, the reduction rate was significantly lower than that of Example 1. Comparative Example 6 showed the lowest reduction rate, which is positively correlated with dry matter digestibility and protein digestibility, confirming the principle that "higher digestibility leads to more complete nutrient absorption and less pollutant emissions in feces."

[0050] Furthermore, lower carbon emissions per unit weight gain indicate better environmental friendliness in the farming process and align more closely with the development trend of low-carbon farming. Compared to the control group, Example 1 showed a significant difference: carbon emissions per unit weight gain were reduced by 16.22%. Based on the comparative data, the multi-synergistic system of the compound enzyme preparation is crucial for achieving low-carbon farming; the absence of any key component will lead to a decrease in the carbon emission reduction effect.

[0051] Table 4. Effects of feeding compound enzyme preparations on nitrogen, phosphorus, and carbon emissions in broilers.

[0052] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A method for preparing a compound enzyme preparation for use in broiler feed, characterized in that, Includes the following steps: S1. Take pollen, crush and sieve it to obtain pollen powder, add acetone, gently stir and reflux at 50℃ for 6 hours, vacuum filter to collect defatted spores, air dry, mix with phosphoric acid solution, stir and reflux at constant temperature, then wash, finally vacuum filter to collect, and dry to obtain spore-coated capsule powder. S2. The outer wall capsule powder of spore-spearin was reconstituted with water to prepare a uniform spore-spearin suspension. The enzyme suspension was slowly added to the spore-spearin suspension, stirred in a constant temperature water bath, and then freeze-dried under vacuum to obtain the core enzyme spore-spearin outer wall capsules. S3. Bile acid analogues, short-chain fatty acid mixtures, and astragalus polysaccharides were dissolved in water and stirred. The core enzyme, sporophytin, was added to the outer wall capsules, emulsified and sheared, and finally hydroxypropyl methylcellulose was added and emulsified and sheared again. Then, the mixture was freeze-dried to obtain a double-layer embedded lyophilized powder. S4. Konjac oligomannose, betaine and betaine phosphate, methacrylic acid copolymer, water were added and stirred to make a mixture, double-layer embedded freeze-dried powder was added to the mixture, homogenized and spray-dried to obtain a preliminary enzyme preparation; S5. Add water to hawthorn powder, stir and dissolve at a constant temperature, filter to obtain a clear hawthorn powder solution, slowly pour sodium alginate solution into the clear hawthorn solution while stirring, then add glycerol and preliminary enzyme preparation and continue stirring, shear emulsify to obtain an emulsion, slowly drip the emulsion into cold calcium chloride solution, let stand, separate the gel particles, wash with water and dry, and sieve to obtain a compound enzyme preparation.

2. The method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S1, the mass ratio of pollen powder to acetone is 1:(3-5); the temperature of gentle stirring and reflux is 45-55℃ for 5-8 hours; the air drying time is 12-16 hours; the concentration of phosphoric acid solution is 70-85 wt.%; and the temperature of constant temperature stirring and reflux treatment is 65-70℃ for 25-30 hours.

3. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: The washing method in step S1 is as follows: after washing with hot water 3-5 times, wash with hot acetone, hot 2M hydrochloric acid, and hot 2M sodium hydroxide in sequence, then wash with hot water 3-5 times, then wash with hot acetone and hot ethanol in sequence, and finally wash with hot water 3-5 times.

4. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S2, the mass ratio of the spore-forming outer wall capsule powder to water is 1:(9-10); the mass ratio of the enzyme suspension to the spore-forming suspension is 1:(2-4); the temperature of the constant temperature water bath stirring is 20-25℃, the rotation speed is 150-250rpm, and the time is 30-50min.

5. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S2, the enzyme suspension is composed of enzymes with the following enzyme activity units: neutral protease 50,000-200,000 U / g, phytase 5,000-20,000 U / g, xylanase 10,000-50,000 U / g, β-glucanase 5,000-30,000 U / g, amylase 5,000-25,000 U / g; and lipase 800-1,000 U / g.

6. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S3, the bile acid analogue is taurocholic acid; the short-chain fatty acid mixture is a mixture of acetic acid, propionic acid, and butyric acid in a mass ratio of (1-2):(1-2):(1-2); the mass ratio of bile acid analogue, short-chain fatty acid mixture, astragalus polysaccharide, hydroxypropyl methylcellulose, and core enzyme spore-forming capsule is (2-3):(2-3):(1-2):(1-2):(12-15); the material-to-liquid ratio of water is 1:(4-8); the stirring and dissolving temperature is 25-30℃, and the stirring speed is 200-300 rpm; the emulsification and shearing speed is 2000-3000 rpm, and the time is 10-20 min.

7. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S4, the methacrylic acid copolymer is L-type; the mass ratio of konjac oligomannose, betaine, betaine phosphate, methacrylic acid copolymer and water is (2-3):(1-2):(1-2):(2-3):(50-80); the ratio of double-layer embedded freeze-dried powder to mixed liquid is 1:(5-8); the homogenization pressure is 10-15 MPa.

8. A method for preparing a compound enzyme preparation for broiler feed according to claim 1, characterized in that: In step S5, the mass ratio of hawthorn powder to water is 1:(15-25); the constant temperature stirring dissolution temperature is 50-55℃, the stirring speed is 300-500rpm, and the time is 20-30min; the concentration of sodium alginate solution is 3-5wt.%; the mass ratio of sodium alginate solution, hawthorn clarified solution, glycerol, and preliminary enzyme preparation is (10-15):(20-40):1:(4-10); the stirring speed is 100-200rpm, and the time is 20-30min; the shear emulsification speed is 2000-4000rpm, and the time is 10-15min; the concentration of cold calcium chloride solution is 2-3wt.%, and the temperature is 4-5℃; the standing time is 10-15min; and the number of water washings is not less than 3.

9. A compound enzyme preparation for use in broiler feed prepared by the preparation method according to any one of claims 1-8.

Images