Preparation method of ruminant rumen bypass protein powder

Abstract

The invention discloses a preparation method of ruminant rumen bypass protein powder, relates to the technical field of rumen bypass protein, and aims to solve the problems that a rumen bypass protein powder coating layer is easy to damage and is dissolved in advance and the ruminant has poor specific absorption in a high-altitude environment. The method comprises the following steps: pretreating concentrated whey protein and soybean protein isolate to obtain a basic protein mixture; preparing a modified shellac solution, and preparing calcium hydroxycitrate-modified shellac microspheres from the modified shellac solution and calcium hydroxycitrate; preparing a gamma-polyglutamic acid-lactobacillus plantarum compound system and a folium cortex eucommiae extract-betaine compound system; the above four components are mixed and dried to obtain the product. The ruminant feed is suitable for high-altitude rumen low-pH environment, improves the stability of a coating layer, gives consideration to energy metabolism regulation, can improve the protein utilization rate and the ruminant production performance, and is suitable for ruminant breeding in high-altitude areas.

Description

Technical Field

[0001] This invention belongs to the field of rumen-protected protein technology, specifically relating to a method for preparing rumen-protected protein powder from ruminants. Background Technology

[0002] The rumen of ruminants is a unique organ for microbial fermentation. Approximately 60% to 80% of ingested protein is degraded into ammonia nitrogen by bacteria and protozoa in the rumen. Only a small amount of undegraded protein can pass through the rumen into the abomasum and small intestine for digestion and absorption. As the farming of ruminants (such as yaks) expands to higher altitudes, and as precision farming demands higher protein utilization efficiency, developing highly efficient rumen-passing protein powder has become crucial for solving the problems of insufficient protein and excessively high degradation rates of conventional proteins in high-altitude farming.

[0003] In high-altitude areas, the low oxygen and low temperature environment significantly alters the physiological characteristics of the rumen in ruminants: on the one hand, the activity of acid-producing bacteria (such as lactic acid bacteria) in the rumen is relatively enhanced, resulting in a rumen pH value that is 0.3 to 0.5 units lower than in plain areas; on the other hand, the rumen peristalsis rate slows down, and the feed retention time is prolonged, further exacerbating the risk of protein degradation in the rumen.

[0004] Although existing technologies can achieve a rumen clearance rate of 60% to 80% in conventional plain environments, there are still insurmountable technical bottlenecks for high-altitude, low-pH rumen environments: (1) Insufficient compatibility of buffer materials: The effective buffer zone of the existing buffer core material is pH 5.5~7.0, which does not match the pH 4.5~5.2 of the rumen at high altitude. The buffering duration is short and it cannot maintain the neutral microenvironment around the coating layer for 2~3 hours when the protein stays in the rumen, resulting in premature dissolution of the coating layer.

[0005] (2) The contradiction between acid resistance and flexibility of wall materials: existing wall materials either have poor acid resistance (sodium alginate has a solubility of over 30% at pH 4.5) or insufficient flexibility (the elongation at break of shellac film is only 5%~8%), which cannot resist the increased chewing intensity caused by the increased energy demand of ruminants at high altitudes, and the covering layer is prone to breakage due to rumination wear.

[0006] (3) Single function and lack of synergistic benefits: Existing technologies only focus on the single goal of protein protection and do not consider the special physiological needs of ruminants at high altitudes. In high-altitude environments, ruminants consume 30% to 40% more energy, but existing rumen protein powders do not have the function of regulating energy metabolism and cannot simultaneously meet the dual needs of protein utilization and energy supplementation, resulting in limited improvement in animal production performance. Summary of the Invention

[0007] The purpose of this invention is to provide a method for preparing rumen-protected protein powder for ruminants, which mainly solves the technical problem that the coating layer of rumen-protected protein powder is easily damaged and dissolved prematurely when fed at high altitudes, resulting in poor specific absorption by ruminants at high altitudes.

[0008] This invention provides a method for preparing rumen-protected protein powder for ruminants, comprising the following steps: S1, pretreating concentrated whey protein and soy protein isolate to obtain a basic protein mixture; S2, preparing a modified shellac solution from natural shellac; S3, mixing calcium hydroxycitrate with the modified shellac solution to prepare calcium hydroxycitrate-modified shellac microspheres; S4, preparing a γ-polyglutamic acid-Lactobacillus plantarum composite system from γ-polyglutamic acid and Lactobacillus plantarum; S5, preparing an Eucommia ulmoides leaf extract-betaine composite system from Eucommia ulmoides leaf extract and betaine; S6, mixing and drying the basic protein mixture, calcium hydroxycitrate-modified shellac microspheres, γ-polyglutamic acid-Lactobacillus plantarum composite system, and Eucommia ulmoides leaf extract-betaine composite system to obtain the rumen-protected protein powder for ruminants.

[0009] This method addresses the challenges of high-altitude environments through a comprehensive process design: basic protein pretreatment → functional wall material preparation → core microsphere synthesis → synergistic system construction → multi-component integration. Basic protein pretreatment achieves amino acid complementarity and process adaptation; modified shellac solution provides acid-resistant and wear-resistant wall materials; calcium hydroxycitrate-modified shellac microspheres construct a buffer-protective core; a γ-polyglutamic acid-Lactobacillus plantarum complex system repairs the intestinal absorption barrier; and an Eucommia ulmoides leaf extract-betaine complex system optimizes rumen fermentation. Finally, through mixing and drying, the various functions are integrated to form a complete technical solution encompassing rumen protection, intestinal absorption, and energy regulation. Calcium hydroxycitrate is a natural component extracted from Garcinia Cambogia, primarily used for weight management, but it has almost no application in the buffering of ruminant feed. Its unique advantage lies in its dual function: firstly, it can synergistically buffer through calcium and citrate ions (buffering range pH 4.5–6.8), with a higher buffering efficiency than sodium bicarbonate; secondly, it can inhibit the activity of lipases in the rumen, reducing energy consumption in ruminants at high altitudes and further improving breeding efficiency. Lac is a natural resin secreted by lac insects and needs to be modified with propylene oxide (to improve acid resistance) before use. Modified lac is almost insoluble in an environment with pH ≤ 5.5, and the membrane has excellent flexibility (elongation at break up to 25%), which can withstand the physical impact of rumination and chewing; however, in the abomasal environment of pH 2.0, the modified groups will rapidly hydrolyze, causing the wall material to disintegrate within 15 minutes, releasing the core material and protein; Within the microspheres formed by the cross-linking of the two materials, the modified shellac wall material slowly degrades in the high-altitude rumen environment (pH 4.5–5.2), causing a slow release of calcium hydroxycitrate from the microspheres. This process precisely covers the residence time of ruminant proteins in the rumen (2–3 hours), preventing interruption of action due to a single release. Direct addition of calcium hydroxycitrate (without coating) would result in a high concentration in the rumen, potentially inhibiting rumen microbial activity. The sustained release from the microspheres maintains a stable HCA-Ca concentration in the rumen at 0.8–1.2 mmol / L, a concentration that effectively regulates metabolism without negatively impacting the rumen microecology. Furthermore, the microspheres exhibit a low release rate after 2 hours in the rumen (pH 4.5); upon entering the abomasum (pH 2.0), the release rate reaches almost complete elimination within 1 hour.

[0010] High-altitude, low-oxygen environments can damage the intestinal mucosa of ruminants, leading to decreased absorption even after rumen-passed proteins enter the small intestine. γ-Polyglutamate (PGA) can form a transparent adhesion membrane (5-8 μm thick) in the intestine, covering the damaged mucosal surface, reducing intestinal epithelial cell apoptosis, and restoring the height of the small intestinal villi to almost normal levels. The energy saved by calcium hydroxycitrate can be preferentially used for intestinal repair, while the enhanced intestinal absorption capacity of PGA further improves the small intestinal absorption rate of rumen-passed proteins, avoiding the waste of protein that passes through the rumen but is not fully absorbed. Calcium hydroxycitrate can promote the reproduction of propionitrile bacteria in the rumen, while this complex system focuses on the linkage between the rumen and gut microbiota: Lactobacillus plantarum can form a symbiotic relationship with propionitrile bacteria in the rumen: the lactic acid produced by Lactobacillus plantarum can be converted into propionic acid by propionitrile bacteria, thereby increasing the production of propionic acid in the rumen on the original basis; after Lactobacillus plantarum that has not colonized the rumen enters the intestine, it can inhibit harmful bacteria such as Escherichia coli, and form a dual defense of microbiota and barrier with the mucosal protection of γ-polyglutamic acid, thereby reducing the intestinal diarrhea that is prone to occur in high-altitude ruminants; The Eucommia ulmoides leaf extract mainly contains chlorogenic acid and eucommia gum, with chlorogenic acid content ≥5% and eucommia gum content ≥10%. Eucommia gum is a natural hydrophobic elastomer that can bind to the dense membrane of modified shellac through physical intercalation. Betaine is a small-molecule amphoteric compound that can be adsorbed onto the surface of microspheres through van der Waals forces. High altitude and low pH inhibit the activity of key fermentation enzymes such as succinate dehydrogenase and acetate kinase in the rumen, leading to a decrease in carbohydrate decomposition efficiency. Chlorogenic acid in the Eucommia ulmoides leaf extract can activate the activity of these enzymes, promote the conversion of carbohydrates in the rumen to volatile fatty acids (VFA), and increase the total amount of VFA compared to the microsphere system alone. Betaine, as a methyl donor, can optimize the methyl metabolism of rumen microorganisms, reduce methane production, and retain more energy as VFA. The energy distribution of calcium hydroxycitrate is combined with the energy generation of this system, thereby increasing the total amount of energy available to ruminants. In addition, the hydrophobicity of Eucommia gum can fill the tiny pores of the modified shellac film, forming a hydrophobic-acid-resistant composite protective layer, further reducing the solubility of calcium hydroxycitrate-modified shellac microspheres in rumen fluid; the moisturizing properties of betaine can delay the drying and aging of the modified shellac film, thus improving the storage stability of the product in high-altitude dry environments.

[0011] Further, S1 specifically includes the following steps: S1.1, mixing the concentrated whey protein and soy protein isolate at a mass ratio of (6.5~7.5):3, then pulverizing and sieving to obtain mixed protein particles; S1.2, adding corn germ powder to the mixed protein particles, wherein the corn germ powder is 12~18% of the mass of the mixed protein particles, mixing evenly, and drying to obtain the basic protein mixture.

[0012] S1.1 utilizes a protein ratio of (6.5~7.5):3, leveraging the complementary amino acid effect of animal protein (whey protein concentrate) and plant protein (soy protein isolate) to compensate for the amino acid deficiencies of single proteins. Pulverization and sieving control particle morphology, laying the foundation for uniform mixing in subsequent steps. S1.2 incorporates corn germ powder, whose crude fat replenishes energy for high-altitude ruminants, while dietary fiber promotes rumen motility. Drying prevents excessive moisture from causing clumping and microbial growth. The basic protein mixture exhibits a balanced amino acid composition, significantly improving protein digestibility compared to conventional single proteins. The addition of corn germ powder balances energy replenishment and rumen function optimization; after drying, the moisture content is <8%, and it remains mold-free for several months of storage, providing a stable and high-quality protein base for subsequent functional component mixing.

[0013] Furthermore, the particle size of the above-mentioned mixed protein particles is less than 180 μm; the water content of the basic protein mixture is less than 8%.

[0014] A particle size of <180 μm increases the specific surface area of ​​the mixed protein particles, which on the one hand reduces the viscosity of rumen chyme at high altitudes, thereby reducing physical friction on the subsequent coating layer, and on the other hand increases the contact area with microspheres and the synergistic system, ensuring uniform mixing; a moisture content of <8% inhibits the adhesion between protein particles, preventing clumping, and at the same time creates a low-moisture environment to prevent microbial growth and protect the activity of subsequent probiotics. Further, S2 specifically includes the following steps: S2.1, mix natural shellac with anhydrous ethanol and heat to 45~55℃ until the natural shellac is completely dissolved to obtain a shellac-ethanol solution, wherein the mass ratio of natural shellac to anhydrous ethanol is 1:(7.5~8.5); S2.2, keep the temperature of S2.1 constant, add propylene oxide dropwise to the shellac-ethanol solution and react for 2~3 h at a dropping rate of 1.2~1.8 mL / min to obtain a mixture, wherein the mass of propylene oxide is 5~7% of the mass of natural shellac; S2.3, adjust the pH of the mixture to 6.0~6.5 with 9~11 wt% calcium carbonate solution, let it stand for 25~35 min, and filter through a 0.45 μm filter membrane to obtain a modified shellac solution.

[0015] S2.1 A 1:(7.5~8.5) ratio of shellac to ethanol and heating at 45~55℃ breaks the hydrogen bonds between shellac molecules, achieving complete dissolution and forming a uniform wall material precursor. In S2.2, propylene oxide undergoes a ring-opening reaction with the carboxyl groups of shellac acid, introducing hydroxyl and ether bonds to construct a stable interpenetrating network and improve acid resistance. In S2.3, calcium carbonate is used to adjust the pH to neutralize acidic residues, and a 0.45μm filter membrane removes impurities, ensuring the purity and activity of the wall material. After modification, the solubility of shellac is significantly reduced after 2 hours in a pH 4.5 environment (high-altitude rumen pH), shear strength is greatly improved, and resistance to ruminant abrasion is significantly enhanced. The pH adjustment and filtration steps remove harmful residues, meet feed safety standards, and avoid irritating the rumen mucosa.

[0016] Further, S3 specifically includes the following steps: S3.1, adding calcium hydroxycitrate to deionized water and stirring to dissolve it to obtain a calcium hydroxycitrate suspension, wherein the mass ratio of calcium hydroxycitrate to deionized water is 1:(1.8~2.2); S3.2, slowly pouring the modified shellac solution into the calcium hydroxycitrate suspension and emulsifying for 40~50 min to obtain a microemulsion; S3.3, adding 1~1.5 wt% calcium chloride solution dropwise to the microemulsion and stirring at 38~42℃ for 1.2~1.8 h to obtain a solidified microsphere solution, wherein the volume of the calcium chloride solution is 13~15% of the volume of the microemulsion, and the dropping rate of the calcium chloride solution is 4~5 mL / min; S3.4, centrifuging the solidified microsphere solution at 4000 rpm for 10~15 min, collecting the lower microsphere precipitate, washing the lower microsphere precipitate 2~3 times with deionized water, and then centrifuging at -50℃ and a vacuum of 10 The calcium hydroxycitrate-modified shellac microspheres were obtained by freeze-drying at Pa for 22-26 h.

[0017] S3.1 Prepare a suspension at a ratio of 1:(1.8~2.2) to ensure uniform dispersion of calcium citrate, whose ions can be slowly released in the rumen at high altitudes to regulate the local pH; S3.2 Emulsify to form a 1.0~1.5 μm microemulsion to achieve uniform encapsulation of the core material by the wall material; S3.3 In S3, calcium chloride forms ionic bonds with the carboxyl groups of shellac to solidify the microspheres, and the reaction is accelerated at 38~42℃ while maintaining the structure; S3.4 Centrifuge and wash to remove impurities, and freeze-dry to avoid shrinkage and cracking of the microspheres.

[0018] After modification with propylene oxide, natural shellac undergoes two key changes in its molecular structure: 1. Retention of active carboxyl groups: The core component of shellac is shellac acid (containing multiple carboxyl groups -COOH). Propylene oxide only reacts with some hydroxyl groups to introduce ether bonds (enhancing flexibility), while retaining a large number of free carboxyl groups (-COOH). These carboxyl groups are the core sites for interaction with calcium hydroxycitrate; 2. Enhanced film density: After modification, shellac molecules form an interpenetrating network structure, resulting in smaller pores (pore size <100nm) after film formation. This allows the shellac to both encapsulate calcium hydroxycitrate particles and slowly release core material ions through the membrane pores.

[0019] Calcium hydroxycitrate is a calcium salt that dissociates into calcium ions (Ca ions) in water. 2+ The carboxyl group (-COOH) of modified shellac will partially dissociate into a carboxyl anion (-COO) in a microemulsion environment (neutral to slightly acidic conditions during preparation). - ): During the emulsification stage (S3.2), when the modified shellac solution is slowly poured into the calcium hydroxycitrate suspension, the -COO groups of the shellac molecules... - Ca that dissociates from calcium hydroxycitrate 2+ Electrostatic attraction occurs, forming -COO -…Ca 2+ Ionic bonds; The subsequent addition of calcium chloride (S3.3) will provide additional Ca. 2+ Not only with shellac's -COO - Furthermore, it can bridge the -COO groups of adjacent shellac molecules. - (Forming - COO) - …Ca 2+ …OOC-), allowing the shellac wall material to more tightly encapsulate the calcium hydroxycitrate particles, preventing core material leakage.

[0020] The strength of these ionic bonds is controllable—strong enough to resist the low pH erosion and rumination friction of the high-altitude rumen (avoiding wall material cracking), but not so tight that the core material cannot be released.

[0021] In addition to carboxyl groups, modified shellac molecules retain a large number of hydroxyl groups (-OH), while calcium hydroxycitrate molecules contain both calcium ions and hydroxyl groups (-OH) and carboxyl groups (-COOH). The -OH group of shellac can form weak hydrogen bonds (-OH…O-) with the -OH and -COOH groups of calcium hydroxycitrate. Although these hydrogen bonds are only 1 / 5 to 1 / 10 the strength of ionic bonds, they can fill the tiny gaps at the interface between the shellac wall material and calcium hydroxycitrate, reduce the porosity of the wall material, further improve the sealing of the microspheres, and prevent the rumen fluid from seeping into the interior of the wall material too early, causing the core material to dissolve prematurely.

[0022] This synergistic physical-chemical cross-linking method also enables precise responses to rumen stabilization and abomasal disintegration: High-altitude rumen (pH 4.5~5.2): The low pH environment will cause the -COO of shellac to... - Reprotonation (reverting to -COOH) slightly weakens the ionic bond strength, but hydrogen bonds still maintain the wall structure; simultaneously, calcium hydroxycitrate slowly releases Ca. 2+ This creates a localized neutral microenvironment around the wall material, further protecting the wall material from dissolution and enabling the slow release of the core material while protecting proteins from passing through the rumen. True stomach (pH 2.0): The highly acidic environment will completely destroy ionic bonds (-COO-). - (Completely protonated), and the hydrogen bonds also disappear with the hydrolysis of the shellac wall material (the ether bonds of the modified shellac break under strong acid). The wall material disintegrates rapidly, releasing the remaining calcium hydroxycitrate and the protected proteins, ensuring absorption in the small intestine.

[0023] Furthermore, the water content of the above-mentioned calcium hydroxycitrate-modified shellac microspheres is ≤3%, and the particle size is 1.0~1.5 μm.

[0024] A moisture content of ≤3% can prevent the microspheres from absorbing moisture and softening during storage, thus preventing damage to the wall material structure; a particle size of 1.0~1.5 μm can avoid phagocytosis by rumen microorganisms and can pass smoothly through the rumen with the chyme, reducing physical retention, while being compatible with basic protein particles (<180 μm) to ensure uniform mixing.

[0025] Further, S4 specifically includes the following steps: S4.1, adding γ-PGA to deionized water to dissolve it and obtain a 9-11 wt% γ-PGA solution; S4.2, adding *Lactobacillus plantarum* to the γ-PGA solution, and dispersing it by ultrasonication to obtain a γ-PGA-*Lactobacillus plantarum* suspension, wherein the mass ratio of *Lactobacillus plantarum* to γ-PGA is 1:(9-11), and the viable count of *Lactobacillus plantarum* is ≥10. 10 CFU / g.

[0026] S4.1 Prepare a 9-11 wt% γ-PGA solution, whose carboxyl and hydroxyl groups can form a protective membrane for the intestinal mucosa and repair intestinal damage caused by high altitude and low oxygen; S4.2 Add highly active Lactobacillus plantarum at a ratio of 1: (9-11), disperse by ultrasonication to avoid aggregation, and encapsulate the bacteria with γ-PGA to protect them from damage in subsequent processes. The high number of viable bacteria ensures intestinal colonization ability.

[0027] Further, S5 specifically includes the following steps: S5.1, mixing Eucommia ulmoides leaf extract and betaine at 150 rpm to obtain a compound dry powder, wherein the mass ratio of Eucommia ulmoides leaf extract to betaine is 3:2; S5.2, adding deionized water to the compound dry powder to obtain a 19~21 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0028] S5.1 Mix at a ratio of 3:2. Chlorogenic acid in Eucommia ulmoides leaf extract activates key enzymes in rumen fermentation, while betaine acts as a methyl donor to reduce methane production. S5.2 Prepare a suspension to prevent dry powder from flying and ensure uniform mixing.

[0029] Further, S6 specifically includes the following steps: S6.1, mixing the basic protein mixture with corn germ oil to obtain a mixture, wherein the mass ratio of the basic protein mixture to the corn germ oil is (9~11):1; S6.2, adding calcium hydroxycitrate-modified shellac microspheres to the mixture and stirring at 110~130 rpm for 10~15 min to obtain a dispersion, wherein the mass ratio of calcium hydroxycitrate-modified shellac microspheres to the basic protein mixture is 1:(12~13); S6.3, sequentially adding the γ-polyglutamic acid-Lactobacillus plantarum complex system and the Eucommia ulmoides leaf extract-betaine complex system to the dispersion and stirring at 110~130 rpm for 12~18 min. A mixed system was prepared by mixing the γ-polyglutamic acid-Lactobacillus plantarum composite system with the hydroxycalcium citrate-modified shellac microspheres at a mass ratio of (1.8~1.9):1, and the eucommia leaf extract-betaine compound system with the hydroxycalcium citrate-modified shellac microspheres at a mass ratio of (1.8~1.9):1. S6.4 After the mixed system naturally cooled to below 30℃, the stirring speed was reduced to 70~90 rpm and stirred for 5~8 min to obtain the final mixture. S6.5 The final mixture was fed into a granulator at 60~70℃ to obtain wet granules. S6.6 The wet granules were dried at 50~60℃ and a wind speed of 1.2~1.8 m / s for 20~30 min to obtain dry granules. S6.7 The dry granules were naturally cooled to room temperature and then passed through a 4~6 mesh sieve to obtain the rumen-protected protein powder from ruminants.

[0030] S6.1 Encapsulate protein with corn germ oil to replenish energy and improve granulation adhesion; S6.2~S6.3 Stir at low speed of 110~130rpm to avoid damaging the structure of microspheres and probiotics, and add to each system in proportion to ensure functional synergy; S6.4 Cool and stir to protect the activity of probiotics; S6.5~S6.6 Low-temperature granulation and drying to preserve heat-sensitive components and control moisture to prevent mold growth; S6.7 Sieve to remove broken particles and ensure uniformity.

[0031] Furthermore, in S6.4 above, the viable count of the final mixture is ≥10. 8 CFU / g; In S6.6 above, the moisture content of the dried granules is ≤12%.

[0032] viable count ≥10 8 CFU / g is the minimum threshold for probiotic colonization in the gut, ensuring that the proportion of beneficial bacteria is ≥30%, so as to exert the functions of degrading microspheres and inhibiting harmful bacteria; the moisture content is ≤12% to balance the needs of high-altitude storage (preventing mold) and particle integrity (preventing breakage), avoiding product defects caused by excessive or insufficient moisture.

[0033] Beneficial effects of the present invention 1. Effectively solves the core problems of premature dissolution of the coating layer and ruminant abrasion in the low pH environment of the rumen at high altitudes: Through the synergistic design of calcium hydroxycitrate and modified shellac microspheres, calcium hydroxycitrate can precisely adapt to the buffering requirements of the rumen pH 4.5~5.2 at high altitudes, slowly releasing ions to maintain a neutral microenvironment around the coating layer and prevent premature dissolution of the coating layer; after being treated with propylene oxide, modified shellac has both excellent acid resistance (almost no dissolution at pH≤5.5) and good flexibility, which can resist the increased chewing intensity of high-altitude ruminants due to increased energy demand, reduce damage to the coating layer caused by physical abrasion, and effectively ensure that protein passes smoothly through the rumen.

[0034] 2. Targeted improvement of poor intestinal absorption in high-altitude ruminants: Relying on the γ-polyglutamic acid-Lactobacillus plantarum complex system, γ-polyglutamic acid can form an adhesion membrane on the surface of the damaged intestinal mucosa, repair the intestinal barrier damage caused by the low oxygen environment of the high altitude, reduce intestinal epithelial cell apoptosis, and restore the normal morphology and absorption function of the small intestinal villi; Lactobacillus plantarum can form a symbiotic relationship with rumen-producing propionibacteria to help optimize the rumen microecology, and can also inhibit the reproduction of harmful bacteria in the intestine. Together with the mucosal protective effect of γ-polyglutamic acid, it forms a dual defense of flora-barrier, avoiding the low absorption efficiency of rumen-passing proteins after entering the small intestine due to the poor intestinal environment.

[0035] 3. To overcome the shortcomings of existing rumen-passing protein powders with limited functions, this product simultaneously meets the dual needs of high-altitude ruminants for protein utilization and energy supplementation: In the Eucommia ulmoides leaf extract-betaine compound system, chlorogenic acid can activate key fermentation enzymes in the high-altitude rumen that are inhibited by low pH, promoting the conversion of carbohydrates into energy substances (volatile fatty acids). Betaine can optimize microbial methyl metabolism and reduce methane production (reducing energy waste). Combined with the inhibitory effect of calcium hydroxycitrate on the activity of rumen lipases (reducing energy consumption), a complete regulatory chain of energy generation, energy conservation, and energy distribution is formed. While ensuring protein passage through the rumen and absorption, this product supplements energy for high-altitude ruminants, avoiding limited performance improvement due to limited functionality. Detailed Implementation

[0036] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention; that is, the described embodiments are merely some embodiments of the invention, and not all embodiments.

[0038] It should be noted that relational terms such as "first" and "second" are used merely to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0039] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0040] Example 1 1. Raw material components Whey protein concentrate (WPC): food grade, protein content ≥80%, dosage 53.03 kg; Soy protein isolate (SPI): food grade, protein content ≥90%, dosage 24.22 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 9.27 kg; Natural shellac: food grade, purity ≥95%, dosage 1.39 kg; Anhydrous ethanol: analytical grade, purity ≥99.5%, dosage 10.42 kg; Propylene oxide: analytical grade, purity ≥99%, dosage 0.07 kg; Calcium carbonate: food grade, purity ≥98%, dosage 0.05 kg; Deionized water (for calcium carbonate solution): none. Specifications and dosage: 0.51 kg (total weight of 9wt% calcium carbonate solution: 0.56 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage 5.27 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage 9.49 kg; Calcium Chloride: Food grade, purity ≥97%, dosage 0.17 kg (total weight of 1wt% calcium chloride solution: 17.0 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage 16.83 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage 1.07 kg; Lactobacillus plantarum: viable count ≥10 10CFU / g, dosage 0.10kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 10.81kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.37kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 0.91kg; Deionized water (for preparing suspension): no specification, dosage 9.70kg; Corn germ oil (CBO): food grade, acid value ≤1.0mg KOH / g, dosage 7.87kg.

[0041] 2. Preparation steps S1: Preparation of basic protein mixture 53.03 kg of concentrated whey protein and 24.22 kg of soy protein isolate were mixed at a mass ratio of 6.5:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 77.25 kg of mixed protein particles. 9.27 kg of corn germ powder (accounting for 12% of the mixed protein particles) was added to the 77.25 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 86.52 kg of basic protein mixture.

[0042] S2: Preparation of modified shellac solution 1.39 kg of natural shellac and 10.42 kg of anhydrous ethanol (mass ratio 1:7.5) were added to a reaction vessel, heated to 45°C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 11.81 kg of shellac-ethanol solution. While maintaining 45°C, 0.07 kg of propylene oxide (5% of natural shellac) was added dropwise to the shellac-ethanol solution at a rate of 1.2 mL / min. After the addition was complete, stirring was continued for 2 h to obtain a mixture. The pH of the mixture was adjusted to 6.0 with a 9 wt% calcium carbonate solution (0.05 kg calcium carbonate + 0.51 kg deionized water), allowed to stand for 25 min, and then filtered through a 0.45 μm aqueous microporous membrane to remove insoluble impurities, yielding 11.80 kg of modified shellac solution.

[0043] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres 5.27 kg of calcium hydroxycitrate and 9.49 kg of deionized water (mass ratio 1:1.8) were added to an emulsification vessel and stirred (500 rpm) for 15 min to obtain 14.76 kg of calcium hydroxycitrate suspension. 11.80 kg of modified shellac solution was slowly poured into the suspension, and emulsification was performed using an emulsifier (3000 rpm) for 40 min to obtain 26.56 kg of microemulsion. 17.0 kg of 1 wt% calcium chloride solution (13% of the microemulsion volume) was added dropwise to the microemulsion at a rate of 4 mL / min. The mixture was heated to 38℃ and stirred (200 rpm) for 1.2 h to obtain 43.56 kg of solidified microsphere solution. The solidified microsphere solution was centrifuged in a high-speed centrifuge (4000 rpm) for 10 min, and the lower layer of microsphere precipitate was collected. The precipitate was washed twice with deionized water (5 kg of water each time) to remove residual calcium chloride. The precipitate was then sent to a freeze dryer (temperature -50℃, vacuum degree 10 Pa) and dried for 22 h to obtain 6.66 kg of calcium hydroxycitric acid-modified shellac microspheres (water content ≤3%, particle size 1.0~1.5 μm).

[0044] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system 1.07 kg of γ-polyglutamic acid and 10.81 kg of deionized water were added to a dissolving vessel and stirred (200 rpm) for 30 min to obtain 11.88 kg of 9 wt% γ-PGA solution. 0.10 kg of *Lactobacillus plantarum* (1:11 mass ratio to γ-PGA) was added to the γ-PGA solution, and the mixture was dispersed using an ultrasonic disperser (300 W) for 10 min to obtain 11.98 kg of a γ-polyglutamic acid-*Lactobacillus plantarum* complex system (viviparous count ≥ 10). 10 CFU / g).

[0045] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system 1.37 kg of Eucommia ulmoides leaf extract and 0.91 kg of betaine were added to a three-dimensional mixer (150 rpm) at a mass ratio of 3:2 and stirred for 15 min to obtain 2.28 kg of compound dry powder. 9.70 kg of deionized water was added to the 2.28 kg of compound dry powder and stirred (200 rpm) for 20 min to obtain 11.98 kg of 19 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0046] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants 86.52 kg of the basic protein mixture and 7.87 kg of corn germ oil (mass ratio 11:1) were added to a double helix mixer (110 rpm) and stirred for 10 min to obtain 94.39 kg of mixture. 6.66 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to the basic protein mixture 1:13) were added to the mixture, and stirring was maintained at 110 rpm for 10 min to obtain 101.05 kg of dispersion. 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex and 11.98 kg of Eucommia ulmoides leaf extract-betaine complex (both with a mass ratio to the microspheres 1:1.8) were added sequentially to the dispersion, and stirring was maintained at 110 rpm for 12 min to obtain 125.01 kg of mixed system. After the mixed system cooled naturally to below 30℃, the stirring speed was reduced to 70 rpm and stirred for 5 min to obtain 125.01 kg of final mixture (viable bacteria count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 60℃) to press, yielding 125.01kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (50℃, 1.2m / s) for 20min to dry, controlling the moisture content to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 6-mesh sieve (3.35mm aperture) to remove fine powder and large particles, yielding 100kg of ruminant rumen-protected protein powder.

[0047] Example 2 Example 2 (between the minimum and the median values) 1. Raw material components Whey protein concentrate (WPC): Food grade, protein content ≥80%, dosage 55.18 kg; Soy protein isolate (SPI): Food grade, protein content ≥90%, dosage 24.93 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 10.89 kg; Natural shellac: Food grade, purity ≥95%, dosage 1.45 kg; Anhydrous ethanol: Analytical grade, purity ≥99.5%, dosage 11.31 kg; Propylene oxide: Analytical grade, purity ≥99%, dosage 0.08 kg; Calcium carbonate: Food grade, purity ≥98%, dosage 0.06 kg; Deionized water (for calcium carbonate solution): No specification. Dosage: 0.59 kg (total weight of 9.5 wt% calcium carbonate solution: 0.65 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage: 5.52 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage: 10.49 kg; Calcium Chloride: Food grade, purity ≥97%, dosage: 0.22 kg (total weight of 1.1 wt% calcium chloride solution: 20.0 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage: 19.78 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage: 1.12 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.11kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 11.19kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.45kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 0.97kg; Deionized water (for preparing suspension): no specification, dosage 10.21kg; Corn germ oil (CBO): food grade, acid value ≤1.0mg KOH / g, dosage 8.24kg.

[0048] 2. Preparation steps S1: Preparation of basic protein mixture 55.18 kg of concentrated whey protein and 24.93 kg of soy protein isolate were mixed at a mass ratio of 6.8:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 80.11 kg of mixed protein particles. 10.89 kg of corn germ powder (accounting for 14% of the mixed protein particles) was added to the 80.11 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 91.00 kg of basic protein mixture.

[0049] S2: Preparation of modified shellac solution 1.45 kg of natural shellac and 11.31 kg of anhydrous ethanol (mass ratio 1:7.8) were added to a reaction vessel, heated to 48 °C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 12.76 kg of shellac-ethanol solution. While maintaining 48 °C, 0.08 kg of propylene oxide (5.5% of the natural shellac) was added dropwise to the shellac-ethanol solution at a rate of 1.5 mL / min. After the addition was complete, stirring was continued for 2.2 h to obtain a mixed solution. The pH of the mixed solution was adjusted to 6.1 using a 9.5 wt% calcium carbonate solution (0.06 kg calcium carbonate + 0.59 kg deionized water), allowed to stand for 28 min, and then filtered through a 0.45 μm aqueous microporous membrane to obtain 12.75 kg of modified shellac solution.

[0050] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres 5.52 kg of calcium hydroxycitrate and 10.49 kg of deionized water (mass ratio 1:1.9) were added to an emulsification vessel and stirred (500 rpm) for 15 min to obtain 16.01 kg of calcium hydroxycitrate suspension. 12.75 kg of modified shellac solution was slowly poured into the suspension, and emulsification was performed using an emulsifier (3000 rpm) for 43 min to obtain 28.76 kg of microemulsion. 20.0 kg of 1.1 wt% calcium chloride solution (occupying 1 / 3 of the microemulsion volume) was added dropwise to the microemulsion at a rate of 4.3 mL / min. 3.5%), heated to 39℃, stirred (200 rpm) for 1.4 h, to obtain 48.76 kg of solidified microsphere solution; put the solidified microsphere solution into a high-speed centrifuge (4000 rpm) for 11 min, collect the lower layer of microsphere precipitate, wash the precipitate twice with deionized water (5 kg of water each time), and send the precipitate into a freeze dryer (temperature -50℃, vacuum degree 10 Pa) for 23 h to obtain 7.08 kg of calcium hydroxycitrate-modified shellac microspheres (water content ≤3%, particle size 1.0~1.5 μm).

[0051] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system 1.12 kg of γ-polyglutamic acid and 11.19 kg of deionized water were added to a dissolving vessel and stirred (200 rpm) for 30 min to obtain 12.31 kg of 9.5 wt% γ-PGA solution. 0.11 kg of *Lactobacillus plantarum* (mass ratio of 1:10.5 to γ-PGA) was added to the γ-PGA solution, and the mixture was dispersed using an ultrasonic disperser (300 W) for 10 min to obtain 12.42 kg of a γ-polyglutamic acid-*Lactobacillus plantarum* complex system (viable count ≥10). 10 CFU / g).

[0052] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system 1.45 kg of Eucommia ulmoides leaf extract and 0.97 kg of betaine were added to a three-dimensional mixer (150 rpm) at a mass ratio of 3:2 and stirred for 15 min to obtain 2.42 kg of compound dry powder. 10.21 kg of deionized water was added to the 2.42 kg of compound dry powder and stirred (200 rpm) for 20 min to obtain 12.63 kg of 19.5 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0053] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants 91.00 kg of the basic protein mixture and 8.24 kg of corn germ oil (mass ratio 10.5:1) were added to a double helix mixer (115 rpm) and stirred for 10 min to obtain 99.24 kg of mixture. 7.08 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to the basic protein mixture 1:12.8) were added to the mixture, and stirring was maintained at 115 rpm for 12 min to obtain 106.32 kg of dispersion. 12.42 kg of γ-polyglutamic acid-Lactobacillus plantarum complex and 12.63 kg of Eucommia ulmoides leaf extract-betaine complex (both with a mass ratio to the microspheres 1:1.83) were added sequentially to the dispersion, and stirring was maintained at 115 rpm for 15 min to obtain 131.37 kg of mixture. After the mixture cooled naturally to below 30°C, the stirring speed was reduced to 75 rpm and stirred for 6 min to obtain 131.37 kg of final mixture (viable bacteria count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 62℃) to press, yielding 131.37kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (52℃, 1.4m / s) for 23min, with the moisture content controlled to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 5-mesh sieve (4.0mm aperture) to obtain 100kg of ruminant rumen-protected protein powder.

[0054] Example 3 1. Raw material components Whey protein concentrate (WPC): Food grade, protein content ≥80%, dosage 57.33 kg; Soy protein isolate (SPI): Food grade, protein content ≥90%, dosage 24.57 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 12.15 kg; Natural shellac: Food grade, purity ≥95%, dosage 1.50 kg; Anhydrous ethanol: Analytical grade, purity ≥99.5%, dosage 12.00 kg; Propylene oxide: Analytical grade, purity ≥99%, dosage 0.09 kg; Calcium carbonate: Food grade, purity ≥98%, dosage 0.07 kg; Deionized water (for calcium carbonate solution): No specification. Dosage: 0.63 kg (total weight of 10wt% calcium carbonate solution: 0.70 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage: 5.77 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage: 11.54 kg; Calcium Chloride: Food grade, purity ≥97%, dosage: 0.28 kg (total weight of 1.25wt% calcium chloride solution: 22.4 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage: 22.12 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage: 1.18 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.12kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 11.62kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.53kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 1.02kg; Deionized water (for preparing suspension): no specification, dosage 10.75kg; Corn germ oil (CBO): food grade, acid value ≤1.0mg KOH / g, dosage 8.60kg.

[0055] 2. Preparation steps S1: Preparation of basic protein mixture 57.33 kg of concentrated whey protein and 24.57 kg of soy protein isolate were mixed at a mass ratio of 7.0:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 81.90 kg of mixed protein particles. 12.15 kg of corn germ powder (accounting for 15% of the mixed protein particles) was added to the 81.90 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 94.05 kg of basic protein mixture.

[0056] S2: Preparation of modified shellac solution 1.50 kg of natural shellac and 12.00 kg of anhydrous ethanol (mass ratio 1:8.0) were added to a reaction vessel, heated to 50 °C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 13.50 kg of shellac-ethanol solution. Maintaining the temperature at 50 °C, 0.09 kg of propylene oxide (6% of natural shellac) was added dropwise to the shellac-ethanol solution at a rate of 1.5 mL / min. After the addition was complete, stirring was continued for 2.5 h to obtain a mixture. The pH of the mixture was adjusted to 6.2 with a 10 wt% calcium carbonate solution (0.07 kg calcium carbonate + 0.63 kg deionized water), allowed to stand for 30 min, and then filtered through a 0.45 μm filter membrane to obtain 13.48 kg of modified shellac solution.

[0057] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres 5.77 kg of calcium hydroxycitrate and 11.54 kg of deionized water (mass ratio 1:2.0) were added to an emulsification vessel and stirred (500 rpm) for 15 min to obtain 17.31 kg of calcium hydroxycitrate suspension. 13.48 kg of modified shellac solution was slowly poured into the suspension, and emulsification was performed using an emulsifier (3000 rpm) for 45 min to obtain 30.79 kg of microemulsion. 22.4 kg of 1.25 wt% calcium chloride solution (accounting for a certain percentage of the microemulsion) was added dropwise to the microemulsion at a rate of 4.5 mL / min. The mixture was heated to 40℃ and stirred (200 rpm) for 1.5 h to obtain 53.19 kg of solidified microsphere solution. The solidified microsphere solution was centrifuged in a high-speed centrifuge (4000 rpm) for 12 min, and the lower layer of microsphere precipitate was collected. The precipitate was washed twice with deionized water (5 kg of water each time). The precipitate was then sent to a freeze dryer (temperature -50℃, vacuum degree 10 Pa) and dried for 24 h to obtain 7.52 kg of calcium hydroxycitrate-modified shellac microspheres (water content ≤3%, particle size 1.0~1.5 μm).

[0058] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system 1.18 kg of γ-polyglutamic acid and 11.62 kg of deionized water were added to a dissolving vessel and stirred (200 rpm) for 30 min to obtain 12.80 kg of 10 wt% γ-PGA solution. 0.12 kg of *Lactobacillus plantarum* (1:10 mass ratio to γ-PGA) was added to the γ-PGA solution, and the mixture was dispersed using an ultrasonic disperser (300 W) for 10 min to obtain 12.92 kg of a γ-polyglutamic acid-*Lactobacillus plantarum* composite system (viable count ≥ 10^6). 10 CFU / g).

[0059] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system 1.53 kg of Eucommia ulmoides leaf extract and 1.02 kg of betaine were added to a three-dimensional mixer (150 rpm) at a mass ratio of 3:2 and stirred for 15 min to obtain 2.55 kg of compound dry powder. 10.75 kg of deionized water was added to the 2.55 kg of compound dry powder and stirred (200 rpm) for 20 min to obtain 13.30 kg of 20 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0060] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants 94.05 kg of the basic protein mixture and 8.60 kg of corn germ oil (mass ratio 10:1) were added to a double helix mixer (120 rpm) and stirred for 10 min to obtain 102.65 kg of mixture. 7.52 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to the basic protein mixture 1:12.5) were added to the mixture, and stirring was maintained at 120 rpm for 12 min to obtain 110.17 kg of dispersion. 12.92 kg of γ-polyglutamic acid-Lactobacillus plantarum complex and 13.30 kg of Eucommia ulmoides leaf extract-betaine complex (both mass ratio to microspheres 1:1.85) were added sequentially to the dispersion, and stirring was maintained at 120 rpm for 15 min to obtain 136.39 kg of mixed system. After the mixed system cooled naturally to below 30℃, the stirring speed was reduced to 80 rpm and stirred for 6 min to obtain 136.39 kg of final mixture (viable bacteria count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 65℃) to press, yielding 136.39kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (55℃, 1.5m / s) for 25min to dry, controlling the moisture content to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 5-mesh sieve (4.0mm aperture) to obtain 100kg of ruminant rumen-protected protein powder.

[0061] Example 4 1. Raw material components Whey protein concentrate (WPC): Food grade, protein content ≥80%, dosage 59.48 kg; Soy protein isolate (SPI): Food grade, protein content ≥90%, dosage 25.06 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 13.41 kg; Natural shellac: Food grade, purity ≥95%, dosage 1.55 kg; Anhydrous ethanol: Analytical grade, purity ≥99.5%, dosage 12.87 kg; Propylene oxide: Analytical grade, purity ≥99%, dosage 0.10 kg; Calcium carbonate: Food grade, purity ≥98%, dosage 0.08 kg; Deionized water (for calcium carbonate solution): No specification. Dosage: 0.68 kg (total weight of 10.5 wt% calcium carbonate solution: 0.76 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage: 6.02 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage: 12.64 kg; Calcium Chloride: Food grade, purity ≥97%, dosage: 0.34 kg (total weight of 1.4 wt% calcium chloride solution: 24.29 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage: 23.95 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage: 1.24 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.13kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 12.05kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.61kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 1.07kg; Deionized water (for preparing suspension): no specification, dosage 11.29kg; Corn germ oil (CBO): food grade, acid value ≤1.0mg KOH / g, dosage 8.96kg.

[0062] 2. Preparation steps S1: Preparation of basic protein mixture 59.48 kg of concentrated whey protein and 25.06 kg of soy protein isolate were mixed at a mass ratio of 7.3:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 84.54 kg of mixed protein particles. 13.41 kg of corn germ powder (accounting for 17% of the mixed protein particles) was added to the 84.54 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 97.95 kg of basic protein mixture.

[0063] S2: Preparation of modified shellac solution 1.55 kg of natural shellac and 12.87 kg of anhydrous ethanol (mass ratio 1:8.3) were added to a reaction vessel, heated to 52 °C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 14.42 kg of shellac-ethanol solution. Maintaining the temperature at 52 °C, 0.10 kg of propylene oxide (6.5% of the natural shellac) was added dropwise to the shellac-ethanol solution at a rate of 1.6 mL / min. After the addition was complete, stirring was continued for 2.6 h to obtain a mixture. The pH of the mixture was adjusted to 6.4 with a 10.5 wt% calcium carbonate solution (0.08 kg calcium carbonate + 0.68 kg deionized water), allowed to stand for 33 min, and then filtered through a 0.45 μm aqueous microporous membrane to obtain 14.40 kg of modified shellac solution.

[0064] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres 6.02 kg of calcium hydroxycitrate and 12.64 kg of deionized water (mass ratio 1:2.1) were added to an emulsification vessel and stirred (500 rpm) for 15 min to obtain 18.66 kg of calcium hydroxycitrate suspension. 14.40 kg of modified shellac solution was slowly poured into the suspension, and emulsification was performed using an emulsifier (3000 rpm) for 47 min to obtain 33.06 kg of microemulsion. 24.29 kg of 1.4 wt% calcium chloride solution (occupying a certain volume of the microemulsion) was added dropwise to the microemulsion at a rate of 4.7 mL / min. 14.5%), heated to 41℃, stirred (200 rpm) for 1.6 h, to obtain 57.35 kg of solidified microsphere solution; put the solidified microsphere solution into a high-speed centrifuge (4000 rpm) for 14 min, collect the lower layer of microsphere precipitate, wash the precipitate twice with deionized water (5 kg of water each time), and send the precipitate into a freeze dryer (temperature -50℃, vacuum degree 10 Pa) for 26 h to obtain 7.96 kg of calcium hydroxycitrate-modified shellac microspheres (water content ≤3%, particle size 1.0~1.5 μm).

[0065] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system 1.24 kg of γ-polyglutamic acid and 12.05 kg of deionized water were added to a dissolving vessel and stirred (200 rpm) for 30 min to obtain 13.29 kg of 10.5 wt% γ-PGA solution. 0.13 kg of *Lactobacillus plantarum* (mass ratio of 1:9.5 to γ-PGA) was added to the γ-PGA solution, and the mixture was dispersed using an ultrasonic disperser (300 W) for 10 min to obtain 13.42 kg of a γ-polyglutamic acid-*Lactobacillus plantarum* composite system (viable count ≥10). 10 CFU / g).

[0066] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system 1.61 kg of Eucommia ulmoides leaf extract and 1.07 kg of betaine were added to a three-dimensional mixer (150 rpm) at a mass ratio of 3:2 and stirred for 15 min to obtain 2.68 kg of compound dry powder. 11.29 kg of deionized water was added to the 2.68 kg of compound dry powder and stirred (200 rpm) for 20 min to obtain 13.97 kg of 20.5 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0067] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants 97.95 kg of the basic protein mixture and 8.96 kg of corn germ oil (mass ratio 9.5:1) were added to a double helix mixer (125 rpm) and stirred for 10 min to obtain 106.91 kg of mixture. 7.96 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to the basic protein mixture 1:12.2) were added to the mixture, and stirring was maintained at 125 rpm for 13 min to obtain 114.87 kg of dispersion. 13.42 kg of γ-polyglutamic acid-Lactobacillus plantarum complex and 13.97 kg of Eucommia ulmoides leaf extract-betaine complex (both with a mass ratio to the microspheres 1:1.88) were added sequentially to the dispersion, and stirring was maintained at 125 rpm for 16 min to obtain 142.26 kg of mixed system. After the mixed system cooled naturally to below 30°C, the stirring speed was reduced to 85 rpm and stirred for 7 min to obtain 142.26 kg of final mixture (viable bacteria count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 68℃) to press, yielding 142.26kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (58℃, 1.7m / s) for 28min, with the moisture content controlled to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 4-mesh sieve (4.75mm aperture) to obtain 100kg of ruminant rumen-protected protein powder.

[0068] Example 5 1. Raw material components Whey protein concentrate (WPC): Food grade, protein content ≥80%, dosage 61.63 kg; Soy protein isolate (SPI): Food grade, protein content ≥90%, dosage 24.65 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 14.70 kg; Natural shellac: Food grade, purity ≥95%, dosage 1.60 kg; Anhydrous ethanol: Analytical grade, purity ≥99.5%, dosage 13.60 kg; Propylene oxide: Analytical grade, purity ≥99%, dosage 0.11 kg; Calcium carbonate: Food grade, purity ≥98%, dosage 0.09 kg; Deionized water (for calcium carbonate solution): No specification. Dosage: 0.73 kg (total weight of 11wt% calcium carbonate solution: 0.82 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage: 6.27 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage: 13.79 kg; Calcium Chloride: Food grade, purity ≥97%, dosage: 0.41 kg (total weight of 1.5wt% calcium chloride solution: 27.33 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage: 26.92 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage: 1.30 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.14kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 12.48kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.69kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 1.13kg; Deionized water (for preparing suspension): no specification, dosage 11.83kg; Corn germ oil (CBO): food grade, acid value ≤1.0mg KOH / g, dosage 9.32kg.

[0069] 2. Preparation steps S1: Preparation of basic protein mixture 61.63 kg of concentrated whey protein and 24.65 kg of soy protein isolate were mixed at a mass ratio of 7.5:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 86.28 kg of mixed protein particles. 14.70 kg of corn germ powder (accounting for 18% of the mixed protein particles) was added to the 86.28 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 100.98 kg of basic protein mixture.

[0070] S2: Preparation of modified shellac solution 1.60 kg of natural shellac and 13.60 kg of anhydrous ethanol (mass ratio 1:8.5) were added to a reaction vessel, heated to 55 °C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 15.20 kg of shellac-ethanol solution. Maintaining the temperature at 55 °C, 0.11 kg of propylene oxide (7% of natural shellac) was added dropwise to the shellac-ethanol solution at a rate of 1.8 mL / min. After the addition was complete, stirring was continued for 3 h to obtain a mixture. The pH of the mixture was adjusted to 6.5 with an 11 wt% calcium carbonate solution (0.09 kg calcium carbonate + 0.73 kg deionized water), allowed to stand for 35 min, and then filtered through a 0.45 μm aqueous microporous membrane to obtain 15.18 kg of modified shellac solution.

[0071] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres 6.27 kg of calcium hydroxycitrate and 13.79 kg of deionized water (mass ratio 1:2.2) were added to an emulsification vessel and stirred (500 rpm) for 15 min to obtain 20.06 kg of calcium hydroxycitrate suspension. 15.18 kg of modified shellac solution was slowly poured into the suspension, and emulsification was carried out in an emulsifier (3000 rpm) for 50 min to obtain 35.24 kg of microemulsion. 27.33 kg of 1.5 wt% calcium chloride solution (accounting for 1 / 3 of the microemulsion volume) was added dropwise to the microemulsion at a rate of 5 mL / min. 15%), heated to 42℃, stirred (200 rpm) for 1.8 h to obtain 62.57 kg of solidified microsphere solution; the solidified microsphere solution was put into a high-speed centrifuge (4000 rpm) for 15 min, the lower layer of microsphere precipitate was collected, the precipitate was washed twice with deionized water (5 kg of water each time), and the precipitate was sent to a freeze dryer (temperature -50℃, vacuum degree 10 Pa) for 26 h to obtain 8.41 kg of calcium hydroxycitrate-modified shellac microspheres (water content ≤3%, particle size 1.0~1.5 μm).

[0072] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system 1.30 kg of γ-polyglutamic acid and 12.48 kg of deionized water were added to a dissolving vessel and stirred (200 rpm) for 30 min to obtain 13.78 kg of 11 wt% γ-PGA solution. 0.14 kg of *Lactobacillus plantarum* (mass ratio 1:9 with γ-PGA) was added to the γ-PGA solution, and the mixture was dispersed using an ultrasonic disperser (300 W) for 10 min to obtain 13.92 kg of a γ-polyglutamic acid-*Lactobacillus plantarum* complex system (viable count ≥ 10^6). 10 CFU / g).

[0073] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system 1.69 kg of Eucommia ulmoides leaf extract and 1.13 kg of betaine were added to a three-dimensional mixer (150 rpm) at a mass ratio of 3:2 and stirred for 15 min to obtain 2.82 kg of compound dry powder. 11.83 kg of deionized water was added to the 2.82 kg of compound dry powder and stirred (200 rpm) for 20 min to obtain 14.65 kg of 21 wt% Eucommia ulmoides leaf extract-betaine suspension.

[0074] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants 100.98 kg of the basic protein mixture and 9.32 kg of corn germ oil (mass ratio 9:1) were added to a double helix mixer (130 rpm) and stirred for 10 min to obtain 110.30 kg of mixture. 8.41 kg of calcium hydroxycitric acid-modified shellac microspheres (mass ratio 1:12 to the basic protein mixture) were added to the mixture, and stirring was maintained at 130 rpm for 15 min to obtain 118.71 kg of dispersion. 13.92 kg of γ-polyglutamic acid-Lactobacillus plantarum complex and 14.65 kg of Eucommia ulmoides leaf extract-betaine complex (both at a mass ratio of 1:1.9 to the microspheres) were added sequentially to the dispersion, and stirring was maintained at 130 rpm for 18 min to obtain 147.28 kg of mixed system. After the mixed system cooled naturally to below 30℃, the stirring speed was reduced to 90 rpm and stirred for 8 min to obtain 147.28 kg of final mixture (viable bacteria count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 70℃) to press 147.28kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (60℃, 1.8m / s) for 30min to control the moisture content to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 4-mesh sieve (4.75mm aperture) to obtain 100kg of ruminant rumen-protected protein powder.

[0075] Comparative Example 1 1. Raw material components Whey protein concentrate (WPC): food grade, protein content ≥80%, dosage 53.03 kg; Soy protein isolate (SPI): food grade, protein content ≥90%, dosage 24.22 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 9.27 kg; Natural shellac: food grade, purity ≥95%, dosage 1.39 kg; Anhydrous ethanol: analytical grade, purity ≥99.5%, dosage 10.42 kg; Calcium carbonate: food grade, purity ≥98%, dosage 0.05 kg; Deionized water (for calcium carbonate solution): no specification, dosage 0.51 kg. 9wt% calcium carbonate solution (total weight 0.56 kg); Calcium hydroxycitrate (HCA-Ca): food grade, purity ≥98%, dosage 5.27 kg; Deionized water (for preparing HCA-Ca suspension): no specification, dosage 9.49 kg; Calcium chloride: food grade, purity ≥97%, dosage 0.17 kg (1wt% calcium chloride solution total weight 17.0 kg); Deionized water (for preparing calcium chloride solution): no specification, dosage 16.83 kg; γ-polyglutamic acid (γ-PGA): food grade, molecular weight 1 million to 2 million Da, dosage 1.07 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.10kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 10.81kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.37kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 0.91kg; Deionized water (for preparing suspension): no specification, dosage 9.70kg; Corn germ oil (CBO): food grade, acid value ≤1.0mgKOH / g, dosage 7.87kg.

[0076] 2. Preparation steps S1: Preparation of basic protein mixture The process was identical to step S1 in Example 1, yielding 86.52 kg of basic protein mixture.

[0077] S2: Preparation of natural shellac solution (unmodified) 1.39 kg of natural shellac and 10.42 kg of anhydrous ethanol (mass ratio 1:7.5) were added to a reaction vessel, heated to 50°C, and stirred (200 rpm) for 30 min until the shellac was completely dissolved, yielding 11.81 kg of natural shellac-ethanol solution. The pH of the solution was adjusted to 6.0 directly with 9 wt% calcium carbonate solution (0.05 kg calcium carbonate + 0.51 kg deionized water), allowed to stand for 25 min, and then filtered through a 0.45 μm aqueous microporous membrane to remove insoluble impurities, yielding 11.80 kg of natural shellac solution.

[0078] S3: Preparation of calcium hydroxycitrate-natural shellac microspheres The procedure was the same as step S3 in Example 1 (except that “modified shellac solution” was replaced with “natural shellac solution”), yielding 6.66 kg of calcium hydroxycitrate-natural shellac microspheres.

[0079] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system The process was completely consistent with step S4 in Example 1, yielding 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex system.

[0080] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system The process was identical to step S5 in Example 1, yielding 11.98 kg of Eucommia ulmoides leaf extract-betaine suspension.

[0081] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants The same steps as in Example 1, S6 (except that “calcium hydroxycitrate-modified shellac microspheres” were replaced with “calcium hydroxycitrate-natural shellac microspheres”), yielded 100 kg of rumen-protected protein powder for ruminants.

[0082] Comparative Example 2 1. Raw material components Whey protein concentrate (WPC): food grade, protein content ≥80%, dosage 53.03 kg; Soy protein isolate (SPI): food grade, protein content ≥90%, dosage 24.22 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 9.27 kg; Natural shellac: food grade, purity ≥95%, dosage 1.39 kg; Anhydrous ethanol: analytical grade, purity ≥99.5%, dosage 10.42 kg; Propylene oxide: analytical grade, purity ≥99%, dosage 0.07 kg; Calcium carbonate: food grade, purity ≥98%, dosage 0.05 kg; Deionized water (for carbonated drinks) Calcium solution: No specification, dosage 0.51 kg (total weight of 9wt% calcium carbonate solution 0.56 kg); Sodium bicarbonate: Food grade, purity ≥99%, dosage 5.27 kg; Deionized water (for preparing sodium bicarbonate suspension): No specification, dosage 9.49 kg; Calcium chloride: Food grade, purity ≥97%, dosage 0.17 kg (total weight of 1wt% calcium chloride solution 17.0 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage 16.83 kg; γ-polyglutamic acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage 1.07 kg; Lactobacillus plantarum: viable count ≥10 10CFU / g, dosage 0.10kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 10.81kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.37kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 0.91kg; Deionized water (for preparing suspension): no specification, dosage 9.70kg; Corn germ oil (CBO): food grade, acid value ≤1.0mgKOH / g, dosage 7.87kg.

[0083] 2. Preparation steps S1: Preparation of basic protein mixture The process was identical to step S1 in Example 1, yielding 86.52 kg of basic protein mixture.

[0084] S2: Preparation of modified shellac solution The modified shellac solution was obtained in exactly the same way as step S2 in Example 1.

[0085] S3: Preparation of sodium bicarbonate-modified shellac microspheres 5.27 kg of sodium bicarbonate and 9.49 kg of deionized water (mass ratio 1:1.8) were added to an emulsification tank and stirred (500 rpm) for 15 min to obtain 14.76 kg of sodium bicarbonate suspension. The subsequent steps were the same as S3.2~S3.4 in Example 1 (only the "calcium hydroxycitrate suspension" was replaced with "sodium bicarbonate suspension") to obtain 6.66 kg of sodium bicarbonate-modified shellac microspheres.

[0086] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system The process was completely consistent with step S4 in Example 1, yielding 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex system.

[0087] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system The process was identical to step S5 in Example 1, yielding 11.98 kg of Eucommia ulmoides leaf extract-betaine suspension.

[0088] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants The same steps as in Example 1, S6 (except that “calcium hydroxycitrate-modified shellac microspheres” were replaced with “sodium bicarbonate-modified shellac microspheres”), yielded 100 kg of rumen-protected protein powder for ruminants.

[0089] Comparative Example 3 1. Raw material components Whey protein concentrate (WPC): food grade, protein content ≥80%, usage 58.95kg; Soy protein isolate (SPI): food grade, protein content ≥90%, usage 26.12kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, usage 9.93kg; Natural shellac: food grade, purity ≥95%, usage 1.39kg; Anhydrous ethanol: analytical grade, purity ≥99.5%, usage 10.42kg; Propylene oxide: analytical grade, purity ≥99%, usage 0.07kg; Calcium carbonate: food grade, purity ≥98%, usage 0.05kg; Deionized water (for calcium carbonate solution): no specification, usage 0.51kg (total weight of 9wt% calcium carbonate solution is 0.56kg); Hydroxy citric acid Calcium chloride (HCA-Ca): Food grade, purity ≥98%, dosage 5.27 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage 9.49 kg; Calcium chloride: Food grade, purity ≥97%, dosage 0.17 kg (total weight of 1wt% calcium chloride solution is 17.0 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage 16.83 kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.37 kg; Betaine: Food grade, anhydrous betaine ≥98%, dosage 0.91 kg; Deionized water (for preparing suspension): No specification, dosage 9.70 kg; Corn germ oil (CBO): Food grade, acid value ≤1.0 mg KOH / g, dosage 8.64 kg.

[0090] 2. Preparation steps S1: Preparation of basic protein mixture 58.95 kg of concentrated whey protein and 26.12 kg of soy protein isolate were mixed at a mass ratio of 6.5:3 and pulverized in an ultrafine grinder (3000 rpm). The mixture was then passed through a 100-mesh sieve (particle size <180 μm) to obtain 85.07 kg of mixed protein particles. 9.93 kg of corn germ powder (accounting for 12% of the mixed protein particles) was added to the 85.07 kg of mixed protein particles and stirred in a double helix mixer (150 rpm) for 20 min until homogeneous. The mixture was then sent to a vacuum dryer (temperature 60℃, vacuum degree -0.09 MPa) and dried for 4 h, controlling the moisture content to <8%, to obtain 95.00 kg of basic protein mixture.

[0091] S2: Preparation of modified shellac solution The modified shellac solution was obtained in exactly the same way as step S2 in Example 1.

[0092] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres The procedure was exactly the same as step S3 in Example 1, yielding 6.66 kg of calcium hydroxycitrate-modified shellac microspheres.

[0093] S4: Preparation of Eucommia ulmoides leaf extract-betaine compound system The process was identical to step S5 in Example 1, yielding 11.98 kg of Eucommia ulmoides leaf extract-betaine suspension.

[0094] S5: Preparation of Rumen-Exposed Protein Powder for Ruminants 95.00 kg of the basic protein mixture and 8.64 kg of corn germ oil (mass ratio 11:1) were added to a double helix mixer (110 rpm) and stirred for 10 min to obtain 103.64 kg of mixture. 6.66 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to basic protein mixture 1:14.26) were added to the mixture, and stirring was maintained at 110 rpm for 10 min to obtain 110.30 kg of dispersion. 11.98 kg of Eucommia ulmoides leaf extract-betaine compound system was added to the dispersion, and stirring was maintained at 110 rpm for 12 min to obtain 122 kg of dispersion. 28 kg of mixing system; after the mixing system has cooled naturally to below 30℃, the stirring speed is reduced to 70 rpm and stirred for 5 min to obtain 122.28 kg of final mixture; the final mixture is fed into a pellet mill (die diameter 3 mm, temperature 60℃) for pressing to obtain 122.28 kg of wet pellets; the wet pellets are fed into a fluidized bed dryer (temperature 50℃, wind speed 1.2 m / s) for drying for 20 min, controlling the moisture content to ≤12%, to obtain 100.0 kg of dried pellets; the dried pellets are cooled naturally to room temperature and passed through a 6-mesh sieve (sieve aperture size 3.35 mm) to obtain 100 kg of ruminant rumen-protected protein powder.

[0095] Comparative Example 4 1. Raw material components Whey protein concentrate (WPC): food grade, protein content ≥80%, dosage 58.95 kg; Soy protein isolate (SPI): food grade, protein content ≥90%, dosage 26.12 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 9.93 kg; Natural shellac: food grade, purity ≥95%, dosage 1.39 kg; Anhydrous ethanol: analytical grade, purity ≥99.5%, dosage 10.42 kg; Propylene oxide: analytical grade, purity ≥99%, dosage 0.07 kg; Calcium carbonate: food grade, purity ≥98%, dosage 0.05 kg; Deionized water (for calcium carbonate solution): none. Specifications and dosage: 0.51 kg (total weight of 9wt% calcium carbonate solution: 0.56 kg); Calcium Hydroxycitrate (HCA-Ca): Food grade, purity ≥98%, dosage 5.27 kg; Deionized water (for preparing HCA-Ca suspension): No specification, dosage 9.49 kg; Calcium Chloride: Food grade, purity ≥97%, dosage 0.17 kg (total weight of 1wt% calcium chloride solution: 17.0 kg); Deionized water (for preparing calcium chloride solution): No specification, dosage 16.83 kg; γ-Polyglutamic Acid (γ-PGA): Food grade, molecular weight 1 million to 2 million Da, dosage 1.07 kg; Lactobacillus plantarum: viable count ≥10 10 CFU / g, dosage 0.10kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 10.81kg; Corn germ oil (CBO): food grade, acid value ≤1.0mgKOH / g, dosage 8.64kg.

[0096] 2. Preparation steps S1: Preparation of basic protein mixture The process was identical to step S1 in Comparative Example 3, yielding 95.00 kg of basic protein mixture.

[0097] S2: Preparation of modified shellac solution The modified shellac solution was obtained in exactly the same way as step S2 in Example 1.

[0098] S3: Preparation of calcium hydroxycitrate-modified shellac microspheres The procedure was exactly the same as step S3 in Example 1, yielding 6.66 kg of calcium hydroxycitrate-modified shellac microspheres.

[0099] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system The process was completely consistent with step S4 in Example 1, yielding 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex system.

[0100] S5: Preparation of Rumen-Exposed Protein Powder for Ruminants 95.00 kg of the basic protein mixture and 8.64 kg of corn germ oil (mass ratio 11:1) were added to a double helix mixer (110 rpm) and stirred for 10 min to obtain 103.64 kg of mixture. 6.66 kg of calcium hydroxycitrate-modified shellac microspheres (mass ratio to basic protein mixture 1:14.26) were added to the mixture, and stirring was maintained at 110 rpm for 10 min to obtain 110.30 kg of dispersion. 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex system was added to the dispersion, and stirring was maintained at 110 rpm for 12 min to obtain 122.28 kg of mixture. After the mixture cooled naturally to below 30°C, the stirring speed was reduced to 70 rpm and stirred for 5 min to obtain 122.28 kg of final mixture (viable count ≥10). 8 The final mixture was fed into a pellet mill (3mm die diameter, 60℃) to press 122.28kg of wet pellets. The wet pellets were then fed into a fluidized bed dryer (50℃, 1.2m / s) for 20min to control the moisture content to ≤12%, yielding 100.0kg of dried pellets. The dried pellets were then allowed to cool naturally to room temperature and passed through a 6-mesh sieve (3.35mm aperture) to obtain 100kg of ruminant rumen-protected protein powder.

[0101] Comparative Example 5 1. Raw material components Whey protein concentrate (WPC): Food grade, protein content ≥80%, dosage 53.03 kg; Soy protein isolate (SPI): Food grade, protein content ≥90%, dosage 24.22 kg; Corn germ powder: crude fat ≥15%, dietary fiber ≥10%, dosage 9.27 kg; Natural shellac: Food grade, purity ≥95%, dosage 1.39 kg; Anhydrous ethanol: Analytical grade, purity ≥99.5%, dosage 10.42 kg; Calcium carbonate: Food grade, purity ≥98%, dosage 0.05 kg; Deionized water (for calcium carbonate solution): No specification, dosage 0 kg. 0.51 kg (total weight of 9wt% calcium carbonate solution: 0.56 kg); Sodium bicarbonate: food grade, purity ≥99%, dosage 5.27 kg; Deionized water (for preparing sodium bicarbonate suspension): no specification, dosage 9.49 kg; Calcium chloride: food grade, purity ≥97%, dosage 0.17 kg (total weight of 1wt% calcium chloride solution: 17.0 kg); Deionized water (for preparing calcium chloride solution): no specification, dosage 16.83 kg; γ-polyglutamic acid (γ-PGA): food grade, molecular weight 1 million to 2 million Da, dosage 1.07 kg; Lactobacillus plantarum: viable count ≥10 10CFU / g, dosage 0.10kg; Deionized water (for preparing γ-PGA solution): no specification, dosage 10.81kg; Eucommia ulmoides leaf extract (EUL): chlorogenic acid ≥5%, eucommia gum ≥10%, dosage 1.37kg; Betaine: food grade, anhydrous betaine ≥98%, dosage 0.91kg; Deionized water (for preparing suspension): no specification, dosage 9.70kg; Corn germ oil (CBO): food grade, acid value ≤1.0mgKOH / g, dosage 7.87kg.

[0102] 2. Preparation steps S1: Preparation of basic protein mixture The process was identical to step S1 in Example 1, yielding 86.52 kg of basic protein mixture.

[0103] S2: Preparation of natural shellac solution (unmodified) The process was identical to step S2 in Comparative Example 1, yielding 11.80 kg of natural shellac solution.

[0104] S3: Preparation of sodium bicarbonate-natural shellac microspheres 5.27 kg of sodium bicarbonate and 9.49 kg of deionized water (mass ratio 1:1.8) were added to an emulsification tank and stirred (500 rpm) for 15 min to obtain 14.76 kg of sodium bicarbonate suspension. The subsequent steps were the same as S3.2~S3.4 in Example 1 (with "modified shellac solution" replaced by "natural shellac solution" and "calcium hydroxycitrate suspension" replaced by "sodium bicarbonate suspension") to obtain 6.66 kg of sodium bicarbonate-natural shellac microspheres.

[0105] S4: Preparation of γ-polyglutamic acid-Lactobacillus plantarum complex system The process was completely consistent with step S4 in Example 1, yielding 11.98 kg of γ-polyglutamic acid-Lactobacillus plantarum complex system.

[0106] S5: Preparation of Eucommia ulmoides leaf extract-betaine compound system The process was identical to step S5 in Example 1, yielding 11.98 kg of Eucommia ulmoides leaf extract-betaine suspension.

[0107] S6: Preparation of Rumen-Exposed Protein Powder for Ruminants The same steps as in Example 1, S6 (except that “calcium hydroxycitrate-modified shellac microspheres” were replaced with “sodium bicarbonate-natural shellac microspheres”), yielded 100 kg of rumen-protected protein powder for ruminants.

[0108] Experimental Example 1 I. Test Subjects Sixty healthy 1.5-year-old yaks (half male and half female, initial weight 180-220 kg) were selected from a plateau region at an altitude of 3500 m. All yaks had no history of rumen disease or parasitic infection, and their rumen pH was measured to be 4.6-5.1 (consistent with the low pH characteristics of the rumen at high altitudes). After 7 days of acclimatization (feeding a basic diet without additives and free access to water), individuals with abnormal weight (<170 kg or >230 kg) and rumen pH deviating from the range of 4.5-5.2 were culled, leaving 50 yaks. These yaks were divided into 10 groups (5 yaks per group), numbered 1-10, corresponding to the experimental diets supplemented with rumen-exposed protein powder from Examples 1-5 and Comparative Examples 1-5, respectively.

[0109] II. Environmental Conditions Experimental location: a high-altitude pasture at 3520m, with an ambient temperature of -5 to 12℃ (a diurnal temperature range of 8 to 10℃), an atmospheric oxygen content of 16.2% to 16.8% (approximately 21% in plains areas), a photoperiod of 10 hours of light (7:00 to 17:00) + 14 hours of darkness, and free ventilation (ammonia concentration < 4 ppm).

[0110] III. Feeding Method The basal diet was uniformly the complete diet for plateau yaks (concentrate to roughage ratio 4:6, crude protein 14%, neutral detergent fiber 45%, metabolizable energy 10.5 MJ / kg, no rumen-protected components). Each experimental diet consisted of the basal diet plus 0.5 wt% of the corresponding group's rumen-protected protein powder. Feeding frequency: twice a day (8:00 and 18:00), single-head fixed-quantity feeding (2.5 kg / 100 kg body weight), ensuring that the feed was consumed within 30 minutes, and the remaining feed was weighed and recorded (to exclude interference from picky eating); drinking water: free access to room temperature snowmelt water throughout the process, and waterers were cleaned and disinfected daily.

[0111] IV. Sample Collection and Testing Methods 1. Sample collection Rumen microsphere collection: On day 14 of the experiment, two yaks from each group were randomly selected to have permanent rumen fistulas installed. 500 mL of rumen fluid was collected at 1 h, 2 h and 3 h after feeding (the time the protein-covered rumen remained in the rumen). The microsphere particles were separated by filtering with a 100-mesh sieve and collected after rinsing three times with physiological saline.

[0112] Simulated wear samples: Take 10g of rumen-protected protein powder granules from each group for later use.

[0113] 2. Detection Method Rumen capsule damage rate: Microsphere morphology was observed using an optical microscope (400x), and the proportion of damaged microspheres (capsule cracking, core material leakage) to the total number of microspheres was counted. Each group was measured in parallel three times, and the average value was taken.

[0114] Rumen pass-through rate determination: Using the chromium (Cr2O3) labeling method, Cr2O3 and rumen pass-through protein powder were mixed at a mass ratio of 1:100 and fed. The contents of the abomasum were collected 6 hours after feeding (the time when the protein entered the abomasum), and the Cr content was determined by atomic absorption spectrophotometer. The rumen pass-through rate was calculated as (total Cr in the abomasum / total Cr in the feed × 100%).

[0115] Simulated rumination wear rate: Protein powder particles were processed for 20 minutes using a reciprocating chewing simulator (frequency 120 times / minute, pressure 50N), passed through a 4-mesh sieve (sieve opening 4.75mm), and the mass of the remaining particles on the sieve was weighed. Wear rate = (initial mass - mass remaining on the sieve) / initial mass × 100%.

[0116] The results are shown in Table 1: Table 1. Acid resistance and abrasion resistance of each coating layer (14-day data)

[0117] According to the results in Table 1: Acid resistance and coating integrity: The rumen coating damage rate of Examples 1-5 was only 7.3%-8.2%, and the rumen pass rate reached 85.6%-87.8%. The core reason is the synergistic design of modified shellac + calcium hydroxycitrate: After treatment with propylene oxide, the modified shellac has a solubility of <5% in the high-altitude rumen environment of pH 4.5-5.1 and an elongation at break of 25%, which is resistant to physical wear; calcium hydroxycitrate slowly releases calcium ions and citrate ions, maintaining a neutral microenvironment of pH 5.0-5.5 around the coating layer and avoiding premature dissolution of the coating.

[0118] Comparative defect verification: Comparative Example 1 (natural shellac, unmodified) had a breakage rate of 35.6% and a rumen pass rate of 62.3%. This is because natural shellac has a solubility of over 30% at pH < 5.5 and poor flexibility (elongation at break of only 6%), making it easily worn down by rumination. Comparative Example 2 (sodium bicarbonate instead of calcium hydroxycitrate) had a rumen clearance rate of 65.8%. However, the buffer zone of sodium bicarbonate was pH 5.5-7.0, which did not match the pH 4.6-5.1 of the rumen at high altitudes. The buffering duration was only 1 hour, which could not maintain the stability of the coating. Comparative Example 5 (natural shellac + sodium bicarbonate) had a breakage rate of 42.7% and a rumen pass rate of 58.5%, exhibiting the dual defects of poor acid resistance and insufficient buffering adaptability, resulting in coating protection failure.

[0119] Wear resistance: The wear rate of Examples 1 and 2-4 was less than 8%, indicating that the flexibility of the modified shellac can resist rumination and chewing. However, the wear rate of Comparative Examples 1 and 5 exceeded 28% due to the high brittleness of natural shellac, further verifying the wear resistance value of the modified shellac.

[0120] Experimental Example 2 I. Test Subjects Similar to Experiment 1 (50 plateau yaks, 10 groups, 5 yaks per group), the experimental period was 5 days of rest after the end of Experiment 1 to ensure that there were no residual microspheres in the rumen, and 1 week after the rumen resumed normal feeding.

[0121] II. Environmental Control and Feeding Methods Environmental control was the same as in Experiment 1; the feeding was changed to “basal diet + 0.5wt% of rumen-protected protein powder of the corresponding group + 0.2wt% casein labeled with sodium fluorescein (tracer protein absorption, stable fluorescence intensity)”, with a single head feeding amount of 2.0kg / 100kg body weight. After feeding, food was restricted but water was allowed, and the feeding time (T0) was recorded.

[0122] III. Sample Collection and Testing Methods 1. Sample collection Intestinal tissue samples: On day 21 of the experiment, two yaks were randomly selected from each group and slaughtered. 1 cm of the middle segment of the small intestine (jejunum) was taken and fixed with 4% paraformaldehyde for later use. Blood samples: Collect 5 mL of jugular vein blood at 2 h, 4 h, and 6 h after feeding (critical period for small intestinal absorption), centrifuge at 3000 rpm for 10 min to separate serum, and set aside for later use; Fecal samples: Collect all feces within 24 hours after feeding, dry to constant weight, crush and pass through a 40-mesh sieve for later use.

[0123] 2. Detection Method Intestinal mucosal repair indicators: Small intestinal villus height: Fixed intestinal tissue was prepared into paraffin sections (5 μm thick), stained with hematoxylin and eosin (HE), and villus height was measured under an optical microscope (400x) (10 villi were measured in each group, and the average value was taken). Apoptosis rate of intestinal epithelial cells: The proportion of apoptotic cells to total epithelial cells was detected by flow cytometry using the TUNEL method (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method).

[0124] Protein absorption efficiency indicators: Serum total protein (TP): Serum TP content (reflecting total protein absorption) is detected by a fully automated biochemical analyzer. Fecal protein residue rate: Fecal crude protein content was determined by the Kjeldahl method, and the residue rate was calculated as (total fecal crude protein / total feed protein) × 100%.

[0125] The results are shown in Table 2: Table 2. Indicators of intestinal mucosal repair and protein absorption efficiency in each group (21-day data)

[0126] According to the results in Table 2: Intestinal mucosal repair effect: The height of small intestinal villi in Examples 1-5 reached 582.3-605.4 μm (significantly higher than 432.6-472.3 μm in Comparative Examples 1-2 and 5), and the epithelial cell apoptosis rate was only 3.6%-4.2%. The core reason is the γ-polyglutamic acid-Lactobacillus plantarum complex system: γ-polyglutamic acid forms a 5-8 μm thick adhesion membrane on the surface of the intestinal mucosa, reducing the damage of the hypoxic environment to epithelial cells; Lactobacillus plantarum inhibits the reproduction of harmful intestinal bacteria (such as Escherichia coli) and reduces the inflammatory response, thus repairing the intestinal barrier with a dual effect.

[0127] Protein absorption efficiency: The serum TP in the example reached 68.5~70.8 g / L, and the fecal protein residue rate was only 7.0%~7.8%. This was because after the intestinal barrier was repaired, the absorption area of ​​the small intestinal villi increased, and the amino acid transport efficiency of rumen-exposed proteins (such as whey protein concentrate and soy protein isolate) was improved. In contrast, the serum TP in Comparative Example 3 (without complex system) was only 60.3 g / L, with a residue rate of 12.5%. Due to the lack of mucosal protection and flora regulation, even if rumen-exposed proteins entered the small intestine, they were still wasted due to insufficient intestinal absorption capacity.

[0128] Comparative examples have shortcomings: Comparative examples 1, 2, and 5 had low rumen clearance rates due to rumen damage and lacked an intestinal repair system. Their serum TP levels were <58 g / L and their residual rates were >14.6%, further validating the necessity of a synergistic design that combines rumen clearance protection with intestinal repair.

[0129] Experimental Example 3 I. Test Subjects Similar to Experiments 1 and 2 (50 plateau yaks, 10 groups, 5 yaks per group), the experimental period was 5 days of rest after Experiment 2 ended, 1 week after resuming normal feeding, and the experiment lasted for 60 days (covering the production performance observation period).

[0130] II. Environmental Control and Feeding Methods Environmental control was the same as in Experiment 1; the diet consisted of “basal diet + 0.5wt% of rumen-protected protein powder corresponding to the group”, with a daily feed intake of 2.5kg / 100kg body weight per head, free access to hay (average daily feed intake of 1.5kg / 100kg body weight), and free access to water.

[0131] III. Sample Collection and Testing Methods 1. Sample collection Rumen fluid: On days 30 and 60 of the experiment, 300 mL of rumen fluid was collected from each group of yaks (same as Experiment 1) through a fistula 2 hours after feeding. The supernatant was collected by centrifugation (4000 rpm, 15 min) and stored at -20℃. Methane gas: On day 60 of the experiment, three yaks were randomly selected from each group and placed in a closed respiratory metabolism chamber (10m³).3 ), and measure 24-hour methane emissions; Production performance records: Weigh the animals once a week and record the initial weight and the final weight at the end of the experiment; for lactating yaks (2 yaks per group), record the milk yield daily and measure the milk protein content (using a fully automated milk analyzer).

[0132] 2. Detection Method Rumen volatile fatty acid (VFA) production: The total VFA, propionic acid, and acetic acid content in rumen fluid were detected by gas chromatography (FFAP capillary column), and the propionic acid ratio was calculated (propionic acid / total VFA × 100%, propionic acid is the main energy source for ruminants). Methane emissions: The methane concentration in the respiratory metabolism chamber is monitored in real time by an infrared methane detector, and the 24-hour methane emissions are calculated in combination with the ventilation volume; Production performance indicators: Weight gain rate = (weight at the end of the test - initial weight) / initial weight × 100%; Milk protein percentage = milk protein content / total solids in milk × 100%.

[0133] The results are shown in Table 3: Table 3. Energy metabolism and production performance indicators for each group (60-day data)

[0134] According to the results in Table 3: Energy metabolism optimization: The total VFA in Examples 1-5 reached 125.6-132.7 mmol / L, the propionic acid ratio was 28.5%-30.2%, and the methane emissions were only 177.5-185.2 L / 24h. The core reason is the synergistic effect of the Eucommia ulmoides leaf extract-betaine compound system and calcium hydroxycitrate. Chlorogenic acid in Eucommia ulmoides leaf extract activates key enzymes in rumen fermentation (such as succinate dehydrogenase), promoting the conversion of carbohydrates into VFAs, with total VFAs increasing by 15% to 25% compared to the control group. Betaine, as a methyl donor, optimizes microbial methyl metabolism, reduces methane production (by 20%–27% compared to the control group), and reduces energy waste; Calcium hydroxycitrate inhibits the activity of rumen lipases, reducing energy consumption caused by excessive fat breakdown and allowing more energy to be used for growth and lactation.

[0135] Improved production performance: The weight gain rate of the example was 12.3%~13.5% and the milk protein rate was 3.25%~3.38%, which was significantly higher than that of the control group (weight gain rate 7.1%~10.2% and milk protein rate 2.82%~3.05%). Due to increased energy production, reduced energy waste and efficient protein absorption, the growth efficiency and milk quality of yaks were improved simultaneously. In contrast, the total VFA of control group 4 (without compound system) was only 115.7 mmol / L, methane emissions were 205.3L, and the weight gain rate was 9.8%, which verifies the key value of the compound system in energy regulation.

[0136] Completeness of the technical solution: This invention addresses the triple pain points of poor protein utilization, weak intestinal absorption, and insufficient energy in plateau ruminants through a full-chain design of rumen protection (modified shellac + calcium hydroxycitrate) → intestinal absorption (γ-polyglutamic acid - Lactobacillus plantarum) → energy regulation (Eucommia ulmoides leaf extract - betaine), resulting in a significant improvement in production performance.

[0137] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, and improvements made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing rumen-protected protein powder from ruminants, characterized in that: Includes the following steps: S1. Take concentrated whey protein and soy protein isolate for material pretreatment to obtain a basic protein mixture; S2. Prepare a modified shellac solution from natural shellac; S3. Mix calcium hydroxycitrate with the modified shellac solution to prepare calcium hydroxycitrate-modified shellac microspheres; S4. Prepare a γ-polyglutamic acid-Lactobacillus plantarum complex system by taking γ-polyglutamic acid and Lactobacillus plantarum. S5. Eucommia ulmoides leaf extract and betaine were used to prepare a compound system of Eucommia ulmoides leaf extract-betaine. S6. The rumen-protected protein powder of the ruminant animal is prepared by mixing the basic protein mixture, calcium hydroxycitrate-modified shellac microspheres, γ-polyglutamic acid-Lactobacillus plantarum complex system and Eucommia ulmoides leaf extract-betaine complex system and drying.

2. The method for preparing rumen-protected protein powder from ruminants according to claim 1, characterized in that: S1 specifically includes the following steps: S1.1 The concentrated whey protein and soy protein isolate are mixed at a mass ratio of (6.5~7.5):3, then pulverized and sieved to obtain mixed protein particles; S1.2 Add corn germ powder to the mixed protein particles, wherein the corn germ powder is 12-18% of the mass of the mixed protein particles, mix well and dry to obtain the basic protein mixture.

3. The method for preparing rumen-protected protein powder for ruminants according to claim 2, characterized in that: The particle size of the mixed protein particles is less than 180 μm; the water content of the basic protein mixture is less than 8%.

4. The method for preparing rumen-protected protein powder from ruminants according to claim 1, characterized in that: S2 specifically includes the following steps: S2.

1. Take natural shellac and anhydrous ethanol, mix them evenly, and heat to 45~55℃ until the natural shellac is completely dissolved to obtain a shellac-ethanol solution. The mass ratio of the natural shellac to the anhydrous ethanol is 1:(7.5~8.5). S2.

2. Keeping the temperature of S2.1 constant, add propylene oxide dropwise to the shellac-ethanol solution and react for 2-3 hours at a dropping rate of 1.2-1.8 mL / min to obtain a mixture. The mass of the propylene oxide is 5-7% of the mass of the natural shellac. S2.3 Adjust the pH of the mixture to 6.0-6.5 with 9-11 wt% calcium carbonate solution, let it stand for 25-35 min, and filter it through a 0.45 μm filter membrane to obtain the modified shellac solution.

5. The method for preparing rumen-protected protein powder for ruminants according to claim 1, characterized in that: S3 specifically includes the following steps: S3.1 Add calcium hydroxycitrate to deionized water and stir to dissolve to obtain a calcium hydroxycitrate suspension, wherein the mass ratio of calcium hydroxycitrate to deionized water is 1:(1.8~2.2). S3.2 Slowly pour the modified shellac solution into the calcium hydroxycitrate suspension and emulsify for 40-50 min to obtain a microemulsion; S3.3 Add 1~1.5 wt% calcium chloride solution to the microemulsion and stir at 38~42℃ for 1.2~1.8 h to obtain a solidified microsphere solution. The volume of the calcium chloride solution is 13~15% of the volume of the microemulsion, and the dropping rate of the calcium chloride solution is 4~5 mL / min. S3.4 Centrifuge the solidified microsphere solution at 4000 rpm for 10-15 min, collect the lower layer of microsphere precipitate, wash the lower layer of microsphere precipitate with deionized water 2-3 times, and then freeze-dry at -50℃ and vacuum degree 10 Pa for 22-26 h to obtain the calcium hydroxycitrate-modified shellac microspheres.

6. The method for preparing rumen-protected protein powder for ruminants according to claim 5, characterized in that: The water content of the calcium hydroxycitrate-modified shellac microspheres is ≤3%, and the particle size is 1.0~1.5 μm.

7. The method for preparing rumen-protected protein powder for ruminants according to claim 1, characterized in that: S4 specifically includes the following steps: S4.1 Add γ-PGA to deionized water to dissolve and obtain a 9~11 wt% γ-PGA solution; S4.

2. Add *Lactobacillus plantarum* to the γ-PGA solution, and disperse by ultrasonication to obtain a γ-PGA-*Lactobacillus plantarum* suspension. The mass ratio of *Lactobacillus plantarum* to γ-PGA is 1:(9~11), and the viable count of *Lactobacillus plantarum* is ≥10. 10 CFU / g.

8. The method for preparing rumen-protected protein powder from ruminants according to claim 1, characterized in that: S5 specifically includes the following steps: S5.

1. Take Eucommia ulmoides leaf extract and betaine and mix them at 150 rpm to obtain a compound dry powder. The mass ratio of Eucommia ulmoides leaf extract to betaine is 3:

2. S5.2 Add deionized water to the compound dry powder to obtain a 19-21 wt% Eucommia ulmoides leaf extract-betaine suspension.

9. The method for preparing rumen-protected protein powder from ruminants according to any one of claims 1, characterized in that: S6 specifically includes the following steps: S6.1 The basic protein mixture is mixed and stirred with corn germ oil to obtain a mixture, wherein the mass ratio of the basic protein mixture to the corn germ oil is (9~11):1; S6.2 Add calcium hydroxycitrate-modified shellac microspheres to the mixture and stir at 110~130 rpm for 10~15 min to obtain a dispersed material. The mass ratio of the calcium hydroxycitrate-modified shellac microspheres to the basic protein mixture is 1:(12~13). S6.

3. Add the γ-polyglutamic acid-Lactobacillus plantarum complex system and the Eucommia ulmoides leaf extract-betaine complex system sequentially to the dispersed material, and stir at 110~130 rpm for 12~18 min to obtain a mixed system. The mass ratio of the γ-polyglutamic acid-Lactobacillus plantarum complex system to the calcium hydroxycitrate-modified shellac microspheres is (1.8~1.9):1, and the mass ratio of the Eucommia ulmoides leaf extract-betaine complex system to the calcium hydroxycitrate-modified shellac microspheres is (1.8~1.9):

1. S6.4 After the mixture has cooled naturally to below 30°C, reduce the stirring speed to 70-90 rpm and stir for 5-8 minutes to obtain the final mixture. S6.

5. The final mixed material is fed into a pellet mill at 60~70℃ and pressed to obtain wet pellets. S6.

6. The wet granules are dried at 50~60℃ and wind speed of 1.2~1.8 m / s for 20~30 min to obtain dried granules; S6.7 After the dried granules are naturally cooled to room temperature, they are passed through a 4-6 mesh sieve to obtain the rumen-protected protein powder of ruminants.

10. The method for preparing rumen-protected protein powder of ruminants according to claim 9, characterized in that: In step S6.4, the viable count of the final mixture is ≥10. 8 CFU / g; in S6.6, the moisture content of the dried granules is ≤12%.

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