A pig feed containing ginsenoside-lipoprotein complex and a preparation method and application thereof

Abstract

The present application belongs to the technical field of livestock feed, and particularly relates to a pig feed containing ginsenoside-lipoprotein complex, a preparation method and application thereof. The feed is composed of a basic daily ration and a ginsenoside-lipoprotein ternary nano additive. The ginsenoside is prepared by four-stage zymolysis of cellulase, pectinase, neutral protease and amylase. The ginsenoside is rich in rare ginsenosides such as Rg3, Rh2 and CK. The carrier is selected from egg yolk phospholipid and egg yolk lipoprotein. In the preparation, the ginseng stem and leaf / residue by-product is first subjected to segmented zymolysis, and then is subjected to ultrasonic-assisted self-assembly, high-pressure homogenization and drying to obtain powder. The ginsenoside extraction rate and the content of rare ginsenosides can be greatly improved. The nano particles are absorbed by lymph to avoid gastric acid degradation and liver first-pass effect. The fattening pigs are fed 30-60 days before slaughter, so that the shear force of lean meat and back fat can be simultaneously reduced, the fat and lean meat quality can be simultaneously tenderized, and the muscle has no ginsenoside residue. The additive amount is extremely low, the raw material cost is low, and the feed is suitable for large-scale antibiotic-free breeding.

Description

Technical Field

[0001] This invention belongs to the field of livestock feed technology, specifically relating to a pig feed containing ginsenoside lipoprotein complex, its preparation method, and its application. Background Technology

[0002] Pork tenderness is a core indicator for consumers to judge the quality of commercial pork. The industry uniformly uses shear force of muscle and fat tissue as a quantitative evaluation standard: the lower the shear force value, the more tender the meat and the better its palatability. In large-scale pig fattening systems, the long-term pursuit of rapid weight gain and high lean meat percentage in breeding and feeding patterns easily leads to problems such as coarse muscle fibers, uneven distribution of intramuscular fat, and coarse and hardened backfat. This results in tough lean meat and greasy, lumpy fat in the finished pork, significantly reducing its commercial value. Currently, the mainstream technical routes for improving pork quality fall into three categories: breeding of superior pig breeds, post-slaughter chilled aging treatment, and regulation with functional feed additives. Among these, plant-based functional feed additives have advantages such as being green and free of chemical drug residues, suitable for large-scale automated farming, and having controllable costs, making them a current research hotspot in the field of antibiotic-reduced and quality-improving pig farming.

[0003] Ginseng stems and leaves, ginseng processing residues, ginseng roots and other ginseng by-products are rich in ginsenosides. These substances have strong antioxidant properties, regulate the body's lipid metabolism, inhibit muscle fiber fibrosis, and relieve oxidative stress. They can improve the physicochemical properties of muscle and adipose tissue from the endogenous level, and are therefore widely developed as functional additives for pig feed.

[0004] CN116870085B discloses a traditional Chinese medicine composition for increasing intramuscular fat content and improving flavor in pork, comprising: ginseng leaves, Panax notoginseng flowers, yam, dried tangerine peel, cinnamon, wolfberry, medicated leaven, and licorice. This composition can be used as a feed additive for pigs, improving pork quality, intramuscular fat content, and flavor. However, by directly mixing dried coarse ginseng leaf powder into feed without any cell wall breaking, enzymatic hydrolysis, or directional conversion extraction processes, most ginsenosides remain locked within the plant fibers in a cell wall-bound state, making them difficult to dissolve and absorb by the digestive tract of livestock. Furthermore, this existing technology only focuses on increasing total intramuscular fat, failing to regulate the density of adipose tissue. Backfat shear force is not significantly reduced, resulting in a coarse, granular texture in the fatty meat. While it can slightly improve the flavor of lean meat, it cannot reduce lean meat shear force, leading to a limited meat quality improvement effect.

[0005] CN105941905A discloses a feed additive and its preparation method for improving pork quality and reducing pig emissions. The additive contains the following ingredients: yam, millet sprouts, chicken gizzard lining, ginseng leaves, turmeric, salvia miltiorrhiza, chuanxiong rhizome, red cimicifuga rhizome, mulberry mistletoe, cibotium barometz, privet fruit, milkweed, cactus, sweet potato flour, garlic, cistanche deserticola, apricot kernel, peanut meal, snake aconite, sodium selenite, and ferrous sulfate. The beneficial effects of this invention are: it contains no antibiotics or hormones, integrates health-promoting, bactericidal, and antiviral functions, establishes and maintains the balance of intestinal flora, significantly improves the pig's immunity, improves and enhances meat quality and flavor, making the meat safe and delicious, significantly increases pig weight while reducing emissions of ammonia and other waste gases, and has a low feed conversion ratio, thus improving the economic benefits for farmers. However, conventional pulverization and mixing or conventional solvent extraction methods can only obtain the free, prototype ginsenosides (Rb1, Rg1, Re, etc.) in the raw materials. Naturally occurring cell wall-bound saponins cannot be effectively released, leading to several problems: ginseng cell walls are composed of a dense network structure formed by the cross-linking of cellulose, pectin, protein, and starch. Current technologies lack a targeted cell wall-breaking and decomposition process, leaving a large amount of bound saponins in the residue, resulting in a low actual dissolution rate of total saponins; the raw materials contain only prototype saponins and lack enzymatic catalytic conversion steps, requiring regulation by Rg3, Rh2, CK, Rk1, etc. Rare ginsenosides with superior meat quality cannot be generated or detected; unencapsulated free ginsenosides are large-molecule glycosides, which are largely degraded by gastric acid and intestinal hydrolytic enzymes in pigs. The remaining active ingredients undergo first-pass metabolism in the liver via the portal vein, and only a very small amount of saponins can reach the target tissues of muscle and fat. High addition amounts are required to produce a slight quality improvement effect, resulting in high costs for breeding. It can slightly increase the water-binding capacity of muscle, but cannot simultaneously reduce the shear force of lean and fat tissues. The technical problems of hard fat and a lot of chewy residue cannot be solved, making it difficult to simultaneously optimize the taste of both lean and fat pork.

[0006] Furthermore, existing extraction methods mostly employ single-enzyme, dual-enzyme combinations, or simple water / alcohol reflux extraction, which can only degrade single components of the cell wall and cannot simultaneously decompose the complex framework of cellulose, pectin, protein, and starch. The few existing technologies using complex enzymatic hydrolysis do not employ a staged pH and temperature-controlled enzymatic hydrolysis strategy; the optimal catalytic conditions of the four enzymes conflict with each other, and enzyme activity inhibits each other, failing to fully release bound saponins and making it difficult to achieve the targeted conversion of prototype saponins to rare ginsenosides. Existing extracts all involve the direct addition of free saponins without phospholipid-lipoprotein complex nanocarriers, meaning saponins cannot avoid degradation in the digestive tract and the first-pass effect in the liver, resulting in extremely low enrichment efficiency in muscle and adipose tissues, requiring high doses to produce a weak effect. Existing technologies only focus on increasing intramuscular fat and improving lean meat color, completely neglecting the optimization of backfat texture and failing to simultaneously reduce shear force in lean and adipose tissues. Most existing technologies directly use ginseng main roots as raw materials, resulting in high raw material procurement costs and rarely utilizing inexpensive ginseng stems, leaves, processing residues, and other byproducts, making large-scale production uneconomical.

[0007] Existing in vitro and animal studies have confirmed that a four-stage enzymatic hydrolysis system involving cellulase, pectinase, neutral protease, and amylase can progressively break down all the complex components of ginseng cell walls, simultaneously releasing bound saponins and catalyzing the desugaring of proto-saponins into rare ginsenosides. Simultaneously, ginsenosides, phospholipids, and lipoproteins self-assemble through hydrophobic interactions and hydrogen bonds to form 80-150nm ternary nanocomposites, which can be absorbed via the lymphatic channels of the pig small intestine, completely bypassing first-pass metabolism in the liver, and significantly increasing the accumulation of saponins in muscle and adipose tissue. However, current technology and industry research have not yet yielded a complete technical solution for combining the four-stage enzymatic hydrolysis of ginsenoside extracts with the ginsenoside-phospholipid-lipoprotein ternary nanocomposites to prepare pig feed for the simultaneous reduction of shear stress in lean and adipose tissues and the simultaneous optimization of tenderness in both lean and fatty pork. Summary of the Invention

[0008] To address the shortcomings of the existing technologies, a pig feed containing ginsenoside lipoprotein complex is provided, along with its preparation method and application. This feed can simultaneously reduce the shear force of lean meat and backfat, while optimizing the tenderness of lean and fatty pork; it also improves the extraction rate of saponins and the conversion rate of rare ginsenosides.

[0009] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is a pig feed containing ginsenoside lipoprotein complex, comprising a basal diet and a ginsenoside lipoprotein complex additive. The ginsenoside lipoprotein complex additive is added to the feed at a rate of 0.01-0.5 mg / kg; the ginsenoside lipoprotein complex is a ternary nanocomposite formed by the self-assembly of ginsenosides, phospholipids, and lipoproteins, and the particle size of the ternary nanocomposite is controlled at 80-150 nm; the ginsenosides are ginseng extracts obtained by quadruple enzymatic hydrolysis of cellulase, pectinase, neutral protease, and amylase.

[0010] The above-mentioned pig feed containing ginsenoside lipoprotein complex, wherein the ginsenosides prepared by the quadruple enzymatic hydrolysis contain at least one of rare ginsenosides Rg3, Rh2, Rg5, Rk1, and CK.

[0011] The aforementioned pig feed containing ginsenoside lipoprotein complex, wherein the phospholipid is selected from one or more of egg yolk lecithin, soybean lecithin, and hydrogenated phospholipids; and the lipoprotein is selected from one or more of egg yolk low-density lipoprotein (LDL), egg yolk high-density lipoprotein (HDL), and egg yolk high-phosphoprotein-derived lipoprotein.

[0012] The aforementioned pig feed containing ginsenoside lipoprotein complex comprises the following basic diet in parts by weight: 50-60 parts corn, 15-20 parts soybean meal, 5-8 parts wheat bran, 1-3 parts fish meal, 1-3 parts soybean oil, 0.8-1.2 parts dicalcium phosphate, 0.3-0.5 parts limestone powder, 0.2-0.4 parts salt, 0.2-0.4 parts L-lysine, 0.1-0.2 parts DL-methionine, and 1-2 parts premix.

[0013] The above-mentioned method for preparing a pig feed containing ginsenoside lipoprotein complex includes the following steps: (1) Quadruple enzymatic hydrolysis extraction: After crushing the raw material containing ginsenosides, add water at a material-to-liquid ratio of 1:8-1:15, adjust the pH to 4.5-6.0, add cellulase, pectinase, neutral protease and amylase for quadruple enzymatic hydrolysis, the enzymatic hydrolysis temperature is 45-55℃, the enzymatic hydrolysis time is 2-6h; after the enzymatic hydrolysis is completed, raise the temperature to 90-100℃ to inactivate the enzyme for 10-15min, centrifuge to collect the supernatant, concentrate, precipitate with alcohol, filter and dry to obtain quadruple enzymatic hydrolyzed ginsenoside extract; (2) Ultrasonic-assisted ternary compounding: The tetra-enzymatic hydrolyzed ginsenoside extract obtained in step (1) is mixed with phospholipids and lipoproteins at a mass ratio of 1:5-20:10-50 and stirred for 30-120 min at 30-50℃ and pH 6.5-7.5. (3) High-pressure homogenization: The mixture obtained in step (2) is subjected to high-pressure homogenization or micro-jet treatment at a pressure of 50-150 MPa for 2-5 cycles. (4) Drying: The homogenized emulsion is spray-dried or freeze-dried to obtain ginsenoside lipoprotein complex additive powder.

[0014] In the above-mentioned method for preparing pig feed containing ginsenoside lipoprotein complex, in step (1), the amounts of the four enzymes added are as follows: cellulase 1000-3000 U / g raw material, pectinase 500-1500 U / g raw material, neutral protease 1000-2500 U / g raw material, and amylase 800-2000 U / g raw material.

[0015] In the above-mentioned method for preparing pig feed containing ginsenoside lipoprotein complex, in step (1), the quadruple enzymatic hydrolysis is a staged enzymatic hydrolysis: in the first stage, the pH is 4.5-5.0, cellulase and pectinase are added, and the enzymatic hydrolysis is carried out at 45-50℃ for 1.5-3h; in the second stage, the pH is adjusted to 6.0-6.5, neutral protease and amylase are added, and the enzymatic hydrolysis is carried out at 50-55℃ for 1.5-3h.

[0016] In the above-mentioned method for preparing a pig feed containing ginsenoside lipoprotein complex, the stirring reaction in step (2) is carried out under ultrasonic assistance, with an ultrasonic power of 200-500W and a frequency of 20-40kHz.

[0017] In the above-mentioned method for preparing a pig feed containing ginsenoside lipoprotein complex, in step (1), the raw material containing ginsenoside is ginseng stems and leaves, ginseng residue or ginseng rootlets.

[0018] The above-mentioned pig feed containing ginsenoside lipoprotein complex involves mixing the ginsenoside lipoprotein complex additive evenly with the basal diet, making it into powdered feed or pelleting it through a pellet mill, and feeding it to fattening pigs in the late stage of fattening 30-60 days before slaughter.

[0019] The beneficial effects of this invention, a pig feed containing ginsenoside lipoprotein complex, its preparation method, and its application, are that this invention does not rely on expensive ginseng root, but uses all industrial waste by-products such as ginseng stems and leaves, ginseng processing residue, and ginseng rootlets as raw materials. These materials are by-products of medicinal material processing, and their market purchase price is only 1 / 8 to 1 / 15 of that of ginseng root. Large-scale continuous production can reduce fixed input of raw materials and solve the technical problem of high feed costs in animal husbandry caused by the large-scale use of ginseng root in existing technologies.

[0020] Ginseng cell walls are composed of a dense network barrier formed by the cross-linking of cellulose, pectin, structural proteins, and starch. Traditional single-enzyme, double-enzyme, and simple water / alcohol extraction methods can only destroy individual cell wall components, leaving a large amount of bound ginsenosides locked in the residue and discarded, resulting in insufficient utilization of the effective components of the raw material. The present invention uses a staged four-stage enzymatic hydrolysis system to degrade four types of skeletal substances stepwise: in the first stage, cellulase and pectinase break down the cellulose and pectin barrier; in the second stage, neutral protease and amylase decompose intracellular proteins and fill in starch, thus breaking down the cell walls intact. Almost all bound and free saponins in the raw material are released, and the content of residual active saponins in the residue is reduced by more than 75%. The quality of active substances produced from the same weight of ginseng by-products is far higher than that of traditional processes, and the comprehensive utilization rate of raw materials is more than doubled.

[0021] Natural ginseng raw materials contain only large-molecule proto-ginsenosides such as Rb1, Rg1, and Re, which have weak activity in regulating lipid metabolism and inhibiting myofibrosis. Rare ginsenosides such as Rg3, Rh2, CK, and Rk1 have even smaller molecular weights and antioxidant and meat quality regulating activities that are 3-5 times stronger than proto-ginsenosides, but their content in natural raw materials is extremely low or even undetectable. This invention's staged enzymatic hydrolysis process can simultaneously catalyze the hydrolysis of glycosidic bonds in proto-ginsenosides, converting low-activity proto-ginsenosides into rare ginsenosides. Traditional water extraction samples show no detectable rare ginsenosides, while the enzymatic hydrolysis product contains a total of 6.8% Rg3, Rh2, Rk1 + CK, increasing the total yield of rare ginsenosides by 3-5 times, thus enhancing the meat quality improvement potential of the additive from the source.

[0022] Existing compound enzymatic hydrolysis technology directly mixes four enzymes for simultaneous reaction. However, the optimal pH and temperature ranges of the four enzymes conflict with each other, and their enzyme activities inhibit each other, resulting in a significant reduction in degradation efficiency. This invention controls the reaction environment in stages: the acidic range degrades fiber pectin, and the near-neutral range decomposes protein starch. Each enzyme works under optimal catalytic conditions, with no activity cancellation, resulting in a full and thorough enzymatic hydrolysis reaction. No additional enzyme dosage is required, saving on enzyme preparation production costs. Attached Figure Description

[0023] Figure 1 A comparison of HPLC chromatograms before and after quadruple enzymatic hydrolysis; among them... Figure 1 -A is the HPLC chromatogram of traditional water extraction. Figure 1 -B is the HPLC chromatogram of the quadruple enzymatic digestion.

[0024] Figure 2 Bar chart showing the effect of different enzyme combinations on the extraction efficiency of ginsenosides.

[0025] Figure 3 The graph shows the effect of different additive amounts on the shear force of pork; among them, Figure 3 -A represents the shear force of lean meat. Figure 3 -B represents the shear force of adipose tissue.

[0026] Figure 4 This is a particle size distribution (DLS) map of the ginsenoside lipoprotein complex. Detailed Implementation

[0027] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments.

[0028] like Figure 1-4 As shown, a pig feed containing ginsenoside lipoprotein complex comprises a basal diet and a ginsenoside lipoprotein complex additive; the basal diet is composed of the following ingredients in parts by weight: 50-60 parts corn, 15-20 parts soybean meal, 5-8 parts wheat bran, 1-3 parts fish meal, 1-3 parts soybean oil, 0.8-1.2 parts dicalcium phosphate, 0.3-0.5 parts limestone, 0.2-0.4 parts salt, 0.2-0.4 parts L-lysine, 0.1-0.2 parts DL-methionine, and 1-2 parts premix.

[0029] Corn provides the primary carbohydrate energy, soybean meal supplies high-quality plant protein, and wheat bran regulates dietary fiber and improves intestinal motility; this protein-energy balance meets the rapid weight gain needs of fattening pigs from 80kg to slaughter, ensuring stable daily weight gain and an excellent feed conversion ratio during the feeding period, without slowing down the slaughter cycle due to nutritional imbalance. A small amount of fishmeal provides animal-derived amino acids and unsaturated fatty acids, compensating for the amino acid deficiency in plant protein, improving the pig's basal metabolic rate, assisting in lipid metabolism regulation, and synergistically enhancing the meat quality improvement effect with ginsenosides. Soybean oil provides essential fatty acids, providing raw materials for the synthesis of intramuscular and subcutaneous fat, and, together with this additive, regulates fat refinement, moderately improving meat aroma and juiciness without producing hard fat. The combination of dicalcium phosphate and limestone precisely meets the needs of bone calcification, preventing later-stage soft hooves and decreased mobility; it ensures normal pig activity, promotes blood circulation, and facilitates the transport and accumulation of saponins in muscle and fat. L-Lysine and DL-methionine are the first and second limiting amino acids for pigs, respectively. These supplement dietary amino acid deficiencies, improve protein deposition efficiency, reduce protein breakdown, and inhibit muscle fiber coarsening and hardening, thus helping to reduce lean meat shear force from a basic nutritional perspective. The compound premix provides antioxidant and metabolic-promoting nutrients such as iron, zinc, selenium, and vitamins A / E, forming a dual antioxidant system with ginsenosides to further reduce post-slaughter muscle oxidative water loss, improve water-holding capacity, and maintain the reddish color of fresh meat. Corn, soybean meal, and wheat bran are all commonly used bulk feed ingredients in the market, with wide procurement channels and minimal price fluctuations. The overall basic diet has low production costs and is suitable for continuous production in large-scale pig farms.

[0030] The ginsenoside lipoprotein complex additive is added to feed at a rate of 0.01-0.5 mg / kg. This microgram-level addition will not dilute core nutrients such as corn, soybean meal, and amino acids, and will not affect weight gain performance, achieving "quality improvement without slowing down." 0.01~0.05 mg: low-cost basic improvement, suitable for ordinary commercial pigs; 0.05~0.3 mg: balanced tenderness and lean meat reduction, recommended range for mainstream premium pigs; 0.3~0.5 mg: ultimate tenderness, specifically for high-end chilled and gift pork. Farmers can flexibly choose according to product positioning, with high cost flexibility. Above 0.3 mg / kg, there is no significant decrease in shear force; 0.5 mg is the upper limit threshold, guiding farmers not to blindly increase the addition amount, saving on additive procurement costs. Existing ginseng powder and ordinary saponin additives are added at tens to hundreds of mg / kg, while the dosage of this invention is only one-thousandth of that, significantly reducing long-term farming costs.

[0031] The ginsenoside lipoprotein complex is a ternary nanocomposite formed by the self-assembly of ginsenosides, phospholipids, and lipoproteins. The phospholipids are selected from one or more of egg yolk lecithin, soybean lecithin, and hydrogenated phospholipids; the lipoproteins are selected from one or more of egg yolk low-density lipoprotein (LDL), egg yolk high-density lipoprotein (HDL), and egg yolk high-phosphoprotein-derived lipoprotein. The particle size of the ternary nanocomposite is controlled at 80-150 nm. The egg yolk / soybean lecithin forms a hydrophobic protective film, isolating the ginsenosides from gastric acid and intestinal hydrolytic enzymes, preventing premature decomposition and loss of ginsenosides in the digestive tract, and significantly reducing activity loss. LDL / HDL has a structure similar to small intestinal chylomicrons; 80-150 nm particles can directly enter the intestinal lymphatic circulation, completely bypassing the portal vein and first-pass metabolism in the liver, significantly increasing the accumulation of ginsenosides in the longissimus dorsi muscle and back fat target tissues, resulting in a several-fold increase in the improvement effect at the same ginsenoside concentration. Phospholipids and yolk lipoproteins are edible food additives, not chemically synthesized carriers, and do not deposit exogenous harmful substances, leaving no residue in the muscle, thus meeting antibiotic-free and green farming standards. 80-150nm is the optimal particle size for lymphatic absorption; particles smaller than 80nm are easily filtered and excreted by the kidneys, while particles larger than 150nm cannot enter chylomicrons. This range ensures maximum absorption efficiency, resulting in stable meat quality improvement effects between batches without fluctuations. Ternary particles spontaneously form through hydrogen bonding and hydrophobic interactions, without introducing organic solvents or cross-linking agents, making production environmentally friendly and leaving no additional impurities in the feed. The ginsenosides are ginseng extracts obtained through a quadruple enzymatic hydrolysis process using cellulase, pectinase, neutral protease, and amylase. The ginsenosides prepared by this quadruple enzymatic hydrolysis contain at least one of the rare ginsenosides Rg3, Rh2, Rg5, Rk1, and CK. Four enzymes degrade cell wall cellulose, pectin, intracellular proteins, and starch respectively, completely opening the dense network barrier and fully releasing bound saponins. The total saponin extraction rate is more than 80% higher than that of traditional water extraction, and the raw material utilization rate is doubled. The enzymatic hydrolysis process catalyzes the desugaring of the original saponins (Rb1, Rg1) to generate rare ginsenosides such as Rg3, Rh2, and CK. These substances have antioxidant, lipid metabolism-regulating, and myofibrosis-inhibiting activities that are 3-5 times stronger than the original saponins, fundamentally improving meat quality. The coexistence of multiple rare ginsenosides acts on both myofibroblast and adipocyte pathways, simultaneously reducing lean meat and fat shear forces, overcoming the limitation of existing technologies that can only improve lean meat individually. Relying solely on biological enzyme catalysis, without the use of acids, alkalis, or chemical glycoside hydrolysis reagents, the extract has no chemical reagent residues, ensuring the safety and compliance of pork fed to the pig.

[0032] A method for preparing a pig feed containing ginsenoside lipoprotein complex includes the following steps: (1) Quadruple enzymatic hydrolysis extraction: After crushing the raw material containing ginsenosides, water is added at a material-to-liquid ratio of 1:8-1:15, the pH is adjusted to 4.5-6.0, and cellulase, pectinase, neutral protease and amylase are added for quadruple enzymatic hydrolysis. The enzymatic hydrolysis temperature is 45-55℃ and the enzymatic hydrolysis time is 2-6h. After the enzymatic hydrolysis is completed, the temperature is raised to 90-100℃ to inactivate the enzyme for 10-15min. The supernatant is collected by centrifugation, concentrated, precipitated with alcohol, filtered and dried to obtain the quadruple enzymatic hydrolyzed ginsenoside extract. The raw material containing ginsenosides is ginseng stems and leaves, ginseng residue or ginseng rootlets, which are waste by-products of ginseng processing. The purchase price is only 1 / 8 to 1 / 15 of that of ginseng main root, which digests the solid waste of medicinal material processing and realizes resource recycling.

[0033] The dosages of the four enzymes added were as follows: cellulase 1000-3000 U / g raw material, pectinase 500-1500 U / g raw material, neutral protease 1000-2500 U / g raw material, and amylase 800-2000 U / g raw material.

[0034] The quadruple enzymatic hydrolysis is a staged process: In the first stage, at pH 4.5-5.0, cellulase and pectinase are added, and hydrolysis is carried out at 45-50℃ for 1.5-3 hours; in the second stage, the pH is adjusted to 6.0-6.5, and neutral protease and amylase are added, and hydrolysis is carried out at 50-55℃ for 1.5-3 hours. Cellulase / pectinase thrives in an acidic environment, while protease / amylase is best suited to a near-neutral environment. The staged reaction ensures that all enzymes operate under optimal catalytic conditions, preventing mutual inhibition. The same amount of enzymes maximizes hydrolysis efficiency and saves on enzyme preparation costs. Too low a solution concentration results in incomplete saponin dissolution; too high a solution concentration leads to high energy consumption and increased production costs. This range balances the extraction rate and concentration energy consumption. The two stages last 1.5-3 hours each, with a total controllable duration of 2-6 hours. Cell wall disruption and saponin conversion can be completed in a short cycle, resulting in fast production turnover and high equipment utilization. The reaction time can be flexibly adjusted according to the degree of raw material breakage. Enzymes are inactivated at 90-100℃, rapidly eliminating all hydrolytic enzymes and preventing continuous degradation and imbalance of saponins in the extract. This ensures stable content of rare ginsenosides in the product and consistent active components across batches. Only water and ethanol are used as solvents; the ethanol can be distilled, recovered, and recycled, resulting in minimal wastewater pollution and environmentally friendly production with no toxic solvent residues.

[0035] (2) Ultrasonic-assisted ternary compounding: The tetra-enzymatically hydrolyzed ginsenoside extract obtained in step (1) is mixed with phospholipids and lipoproteins at a mass ratio of 1:5-20:10-50, and stirred for 30-120 min at 30-50℃ and pH 6.5-7.5. The stirring reaction is carried out under ultrasonic assistance with an ultrasonic power of 200-500W and a frequency of 20-40kHz. The ratio of saponins:phospholipids:lipoproteins ensures a complete hydrophobic encapsulation structure. Sufficient phospholipids form a protective film, and sufficient lipoproteins achieve lymphatic targeting. Too low a ratio easily leads to free saponins, while too high a ratio wastes the carrier. The mild reaction at 30~50℃ protects the active saponins. High temperatures will destroy the structure of rare ginsenosides. The low temperature range ensures stable self-assembly and no degradation loss of active components. The neutral pH of 6.5~7.5 is suitable for the stability of phospholipids and lipoproteins. Excessive acidity or alkalinity will lead to hydrolysis of phospholipids and denaturation of lipoproteins. The neutral environment ensures the integrity of the carrier structure. Large aggregates are broken down by 200-500W, 20-40kHz ultrasound, promoting full molecular contact among the three components and spontaneously forming spherical nanoparticles. This reduces the need for subsequent homogenization pressure and saves equipment energy. The reaction time is 30-120 minutes; a shorter time can prepare the basic complex, while a longer time improves particle uniformity. Factories can adjust the stirring and ultrasound time according to production capacity requirements.

[0036] (3) High-pressure homogenization: The mixture obtained in step (2) is subjected to high-pressure homogenization or micro-jet treatment at a pressure of 50-150 MPa for 2-5 cycles. The particle size is precisely controlled to 80-150 nm at 50-150 MPa. Low pressure cannot break up large agglomerates, while excessive pressure will tear the phospholipid membrane and damage the nanostructure. This pressure range precisely controls the particles to the optimal size for lymphatic absorption. Cycling 2-5 times improves the dispersion and reduces the PDI. Multiple homogenizations eliminate large particle impurities, reduce the particle distribution width, and ensure uniform dispersion of powder during mixing, preventing local concentration exceedances in the feed.

[0037] (4) Drying: The homogenized emulsion is spray-dried or freeze-dried to obtain ginsenoside lipoprotein complex additive powder. The solid powder is convenient for storage, transportation, and mixing. The dried powder has low moisture content and its activity does not significantly decrease after 6 months of sealed storage at room temperature, eliminating the need for cold chain transportation. It exhibits excellent dispersibility when mixed with granular / powdered feed and is compatible with fully automatic mixing equipment. The dry powder has no risk of mold growth, extends the shelf life of the finished feed, and reduces storage losses.

[0038] Application of pig feed containing ginsenoside lipoprotein complex: The ginsenoside lipoprotein complex additive is mixed evenly with the basal diet to form powdered feed or pelleted. Powdered feed is suitable for small-scale pig farms and individual farmers who mix their own feed; pelleted feed has good palatability, reduces feed waste, and is suitable for large-scale fully automated feeding lines. Only a conventional feed mixer is needed for even mixing, without increasing investment in farm equipment. It is used for feeding fattening pigs in the late fattening stage (30-60 days before slaughter). No need to add it throughout the piglet's life; only add the additive 1-2 months before slaughter, shortening the usage cycle and significantly reducing overall farm investment compared to long-term additive programs. Intervention at this stage, after pigs reach 80kg, during the fat deposition and muscle shaping phase, maximizes the control of fat refinement and inhibits muscle fiber coarsening, resulting in optimal meat quality improvement. In the early stages, piglets' metabolism is not yet fully developed, and the benefits of adding the additive are extremely low. The piglet stage focuses on skeletal and organ development; this additive targets meat tenderization, and adding it only in the late fattening stage does not interfere with the normal growth control program for piglets. Example

[0039] The preparation of ginsenoside extract by four-stage enzymatic hydrolysis includes the following steps: Take 1000g of dried ginseng stems and leaves, pulverize and pass through a 40-mesh sieve. Add 10L of deionized water at a material-to-liquid ratio of 1:10 and stir well. Adjust the pH to 4.8, add 2000U / g cellulase and 1000U / g pectinase, and hydrolyze in a 48℃ water bath for 2 hours; then adjust the pH to 6.2, add 1800U / g neutral protease and 1500U / g amylase, and hydrolyze in a 52℃ water bath for 2 hours. After hydrolysis, heat to 95℃ for 15 minutes to inactivate the enzymes, cool to room temperature, and centrifuge (8000rpm, 10 minutes), collecting the supernatant. Concentrate the supernatant under vacuum to 1 / 5 of its original volume, add 3 times its volume of 95% ethanol for precipitation, let stand at 4℃ for 12 hours, filter, recover the ethanol from the filtrate, and freeze-dry to obtain the tetrahydrolyzed ginsenoside extract.

[0040] HPLC detection conditions: C18 reversed-phase column (250 mm × 4.6 mm, 5 μm); mobile phase: acetonitrile-water gradient elution; detection wavelength: 210 nm; flow rate: 1.0 mL / min; column temperature: 30 ℃. External standard method was used, with ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rd, Rg3, Rh2, Rk1, and CK standards as references.

[0041] The test results are shown in the table below:

[0042] This embodiment uses inexpensive ginseng stems and leaves as raw materials, eliminating the need for expensive ginseng roots. The four-stage enzymatic hydrolysis completely degrades the cell wall cellulose, pectin, protein, and starch complex framework, increasing the total saponin extraction rate by 80.7% compared to traditional water extraction. The residual saponins in the residue are significantly reduced, and the production of active substances from the same amount of raw materials is doubled, solving the problems of low raw material utilization and waste of a large amount of effective ingredients in traditional extraction.

[0043] Traditional water extraction can only obtain low-activity prototype saponins such as Rg1 and Rb1, with no rare ginsenosides detected. By segmented enzymatic hydrolysis and catalytic desugaring of glycosidic bonds, four types of highly active rare ginsenosides, Rg3, Rh2, Rk1, and CK, were successfully generated, with a total content of 6.8%. The yield of rare ginsenosides is more than 8 times higher than that of traditional processes, which greatly improves the meat quality control capabilities of additives.

[0044] This embodiment matches the optimal reaction conditions of four enzymes in two stages. Cellulase and pectinase are fully utilized in the acidic low-temperature range, while protease and amylase are active in the near-neutral high-temperature range. There is no activity inhibition problem in the mixed reaction of multiple enzymes, and there is no need to increase the amount of enzyme preparation added, thus reducing the extraction and production cost.

[0045] Extraction uses only water and ethanol. The ethanol after precipitation can be distilled and reused, resulting in minimal pollutants in the waste liquid, which meets the environmental protection requirements for the production of green feed additives. Example

[0046] The preparation of ginsenoside lipoprotein complex additives includes the following steps: 100g of the tetra-enzymatically hydrolyzed ginsenoside extract prepared in Example 1 was dissolved in water until the solid content reached 10%. 600g of egg yolk lecithin and 2500g of egg yolk low-density lipoprotein (LDL) were added, and the pH was adjusted to 7.0. The mixture was stirred and reacted at 40℃ and ultrasonic power of 300W (frequency 28kHz) for 60 min. The resulting mixture was then homogenized using a high-pressure homogenizer at 100MPa for 3 cycles. Finally, it was spray-dried (inlet air temperature 180℃, outlet air temperature 80℃) to obtain ginsenoside lipoprotein complex additive powder.

[0047] Test results Laser particle size analyzer (Malvern Zetasizer NanoZS) showed that the average particle size of the composite was 105 nm, with a PDI of 0.18. Transmission electron microscopy (TEM) revealed that the composite consisted of spherical nanoparticles, uniformly dispersed, without obvious agglomeration, and the particle size fell entirely within the target range of 80–150 nm.

[0048] Phospholipids form a hydrophobic protective film on the outer layer of the particles, which isolates the stomach acid and intestinal hydrolytic enzymes of pigs, preventing the premature decomposition and loss of ginsenosides, and greatly reducing the loss of active ingredients in the digestive tract.

[0049] The 105nm nanoparticles match the absorption channels of chylomicrons in the small intestine and are not metabolized by the liver via the portal vein. The active saponins can be selectively enriched in muscle and adipose tissues. At the same dosage, the effective concentration in vivo is much higher than that of free saponin extracts.

[0050] The product has a PDI of only 0.18 and no particle agglomeration. The spray-dried powder has strong stability when stored in a sealed container at room temperature. When mixed, it can be evenly dispersed in complete feed without local concentrations that are too high or too low. It is suitable for continuous pelleting and mixing production lines in feed mills.

[0051] Ultrasonic stirring, high-pressure homogenization, and spray drying are all standard equipment in the plant extract and feed additive industries. No special large-scale equipment is required, and small and medium-sized processing plants can all implement production there. Example

[0052] Preparation of basic feed for fattening pigs: Basic diet formula (by weight): 55 parts corn, 18 parts soybean meal, 6 parts wheat bran, 2 parts fish meal, 2 parts soybean oil, 1.0 part dicalcium phosphate, 0.4 parts limestone powder, 0.3 parts salt, 0.3 parts L-lysine, 0.15 parts DL-methionine, and 1.5 parts premix.

[0053] The ginsenoside lipoprotein complex additive prepared in Example 2 was mixed evenly with the basal diet at an addition rate of 0.1 mg / kg (calculated as ginsenosides), and granulated by a ring die pellet mill (pore size 4.0 mm) to obtain a fattening pig feed containing ginsenoside lipoprotein complex.

[0054] With a trace addition ratio of only 0.1mg / kg, it will not dilute the core nutrients such as corn, soybean meal, and amino acids. After feeding, it will not affect the daily weight gain or feed conversion ratio of pigs, achieving quality improvement without sacrificing growth performance.

[0055] The basic diet's energy, protein, amino acid, and mineral ratios are aligned with the growth standards for fattening pigs 30-60 days before slaughter. When combined with the additives of this invention, normal weight gain and meat quality improvement can be achieved simultaneously.

[0056] It can be fed directly as powdered feed or processed into pellets, and is suitable for both small-scale farmers who mix their own feed and large-scale pig farms with fully automated feeding equipment. Example

[0057] Experimental Design: Ninety Duroc × Landrace × Large White crossbred fattening pigs weighing approximately 80 kg were randomly divided into three groups of 30 pigs each: Control group: fed a basal diet; Water-extracted group: fed a basal diet plus 0.1 mg / kg of conventionally extracted water-extracted ginsenoside lipoprotein complex; Enzymatic hydrolysis group: fed a basal diet plus 0.1 mg / kg of quadruple-enzymatically hydrolyzed ginsenoside lipoprotein complex (prepared in Example 2). The pre-trial period was 7 days, and the formal trial period was 45 days. Pigs had free access to feed and water and were managed using standard feeding and management methods.

[0058] On the last day of the experiment, 10 pigs were randomly selected from each group and slaughtered. The longissimus dorsi muscle (lean meat) and backfat (adipose tissue) were harvested. Shear force was measured using the Warner-Bratzler shear method, and ginsenoside residues in the muscle and abdominal fat were also detected. The results are shown in the table below:

[0059] Lean meat improvement: Compared with the blank control group, the shear force of lean meat in the enzymatic hydrolysis group was reduced by 23.0%; effectively inhibiting muscle fiber fibrosis and solving the technical problems of commercial lean meat becoming hard and dry.

[0060] Improved backfat: Compared to the control group, the shear force of backfat decreased by 38.8%; the fat cells were refined and uniform, and the fat was free of coarse particles and greasy lumps, which greatly enhanced the value of high-end pork products.

[0061] No ginsenosides were detected in the edible lean meat tissue. Only trace amounts of ginsenosides (0.02 μg / g) were present in the abdominal fat. Since most of the abdominal fat was removed during cooking, there was no risk of human ingestion. The results met all domestic standards for the detection of residues in fresh meat and exported livestock and poultry meat.

[0062] The three groups had the same dosage of added ginsenosides, with differences only in extraction process and active components. This directly demonstrates that the synergistic effect of quadruple enzymatic hydrolysis and nanocarrier can significantly improve meat quality. Example

[0063] Experimental Design: Using the basal diet of Example 3, five additive gradients were set up: 0.01, 0.05, 0.1, 0.3, and 0.5 mg / kg (calculated as ginsenosides). A blank control group received no additives. All diets were fed uniformly for 45 days, and the shear force of lean meat and adipose tissue was measured. The results are shown in the table below:

[0064] When the addition amount is below 0.01 mg / kg, the improvement in meat quality is weak and has no practical application value; in the range of 0.05~0.3 mg / kg, the shear force decreases significantly and the quality improvement effect is stable; when the addition amount exceeds 0.3 mg / kg, the shear force of lean meat and fat does not decrease significantly further, and there is no dose-dependent synergistic effect. Farmers can choose the 0.05~0.3 mg / kg range according to the market positioning of pork, balancing meat quality effect and breeding input costs.

[0065] An addition of 0.1 mg / kg can reduce lean meat shear force by 23.0% and fat shear force by 38.8%. Further increasing the addition will only slightly improve the improvement effect, but will increase the purchase cost of the additive. It has the highest overall cost performance and is the recommended standard addition dosage for large-scale farms.

[0066] Significant quality improvement can be achieved by adding the additive 30-60 days before slaughter, eliminating the need for feeding piglets throughout their lives, greatly shortening the additive usage cycle, and further reducing overall breeding costs.

[0067] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A pig feed containing ginsenoside lipoprotein complex, characterized in that: It contains a basic diet and a ginsenoside lipoprotein complex additive; The ginsenoside lipoprotein complex additive is added to the feed at a rate of 0.01-0.5 mg / kg; the ginsenoside lipoprotein complex is a ternary nanocomposite formed by the self-assembly of ginsenosides, phospholipids, and lipoproteins, and the particle size of the ternary nanocomposite is controlled at 80-150 nm; the ginsenosides are ginseng extracts obtained by quadruple enzymatic hydrolysis of cellulase, pectinase, neutral protease, and amylase.

2. The pig feed containing ginsenoside lipoprotein complex according to claim 1, characterized in that, The ginsenosides prepared by the quadruple enzymatic hydrolysis contain at least one of the rare ginsenosides Rg3, Rh2, Rg5, Rk1, and CK.

3. The pig feed containing ginsenoside lipoprotein complex according to claim 2, characterized in that, The phospholipids are selected from one or more of egg yolk lecithin, soybean phospholipids, and hydrogenated phospholipids; the lipoproteins are selected from one or more of egg yolk low-density lipoprotein (LDL), egg yolk high-density lipoprotein (HDL), and egg yolk high-phosphoprotein-derived lipoproteins.

4. The pig feed containing ginsenoside lipoprotein complex according to claim 3, characterized in that, The basic diet consists of the following ingredients by weight: 50-60 parts corn, 15-20 parts soybean meal, 5-8 parts wheat bran, 1-3 parts fish meal, 1-3 parts soybean oil, 0.8-1.2 parts dicalcium phosphate, 0.3-0.5 parts limestone powder, 0.2-0.4 parts salt, 0.2-0.4 parts L-lysine, 0.1-0.2 parts DL-methionine, and 1-2 parts premix.

5. A method for preparing a pig feed containing ginsenoside lipoprotein complex according to any one of claims 1-4, characterized in that: Includes the following steps: (1) Quadruple enzymatic hydrolysis extraction: After crushing the raw material containing ginsenosides, add water at a material-to-liquid ratio of 1:8-1:15, adjust the pH to 4.5-6.0, add cellulase, pectinase, neutral protease and amylase for quadruple enzymatic hydrolysis, the enzymatic hydrolysis temperature is 45-55℃, the enzymatic hydrolysis time is 2-6h; after the enzymatic hydrolysis is completed, raise the temperature to 90-100℃ to inactivate the enzyme for 10-15min, centrifuge to collect the supernatant, concentrate, precipitate with alcohol, filter and dry to obtain quadruple enzymatic hydrolyzed ginsenoside extract; (2) Ultrasonic-assisted ternary compounding: The tetra-enzymatic hydrolyzed ginsenoside extract obtained in step (1) is mixed with phospholipids and lipoproteins at a mass ratio of 1:5-20:10-50 and stirred for 30-120 min at 30-50℃ and pH 6.5-7.

5. (3) High-pressure homogenization: The mixture obtained in step (2) is subjected to high-pressure homogenization or micro-jet treatment at a pressure of 50-150 MPa for 2-5 cycles. (4) Drying: The homogenized emulsion is spray-dried or freeze-dried to obtain ginsenoside lipoprotein complex additive powder.

6. The method for preparing a pig feed containing ginsenoside lipoprotein complex according to claim 5, characterized in that, In step (1), the amounts of the four enzymes added are as follows: cellulase 1000-3000 U / g raw material, pectinase 500-1500 U / g raw material, neutral protease 1000-2500 U / g raw material, and amylase 800-2000 U / g raw material.

7. The method for preparing a pig feed containing ginsenoside lipoprotein complex according to claim 6, characterized in that, In step (1), the quadruple enzymatic hydrolysis is a staged enzymatic hydrolysis: in the first stage, the pH is 4.5-5.0, cellulase and pectinase are added, and the enzymatic hydrolysis is carried out at 45-50℃ for 1.5-3 hours; in the second stage, the pH is adjusted to 6.0-6.5, neutral protease and amylase are added, and the enzymatic hydrolysis is carried out at 50-55℃ for 1.5-3 hours.

8. The method for preparing a pig feed containing ginsenoside lipoprotein complex according to claim 7, characterized in that, The stirring reaction described in step (2) is carried out under ultrasonic assistance, with an ultrasonic power of 200-500W and a frequency of 20-40kHz.

9. The method for preparing a pig feed containing ginsenoside lipoprotein complex according to claim 8, characterized in that, In step (1), the raw materials containing ginsenosides are ginseng stems and leaves, ginseng residue, or ginseng roots.

10. The application of a pig feed containing ginsenoside lipoprotein complex according to any one of claims 1-4, characterized in that: The ginsenoside lipoprotein complex additive is mixed evenly with the basal diet and made into powdered feed or pelleted by a pellet mill for feeding fattening pigs in the late stage of fattening 30-60 days before slaughter.

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