Application of plant-derived steroidal derivative and feed additive

Abstract

The application relates to the technical field of feed additives, in particular to application of a plant-derived steroidal derivative and a feed additive; the plant-derived steroidal derivative with a molecular structure as shown in formula 1 is applied in an additive of livestock and poultry feed; in formula 1, R1 and R2 groups respectively include a carbon-carbon single bond or a carbon-carbon double bond; and the R3 group includes oxygen or a hydroxyl group. The application uses the plant-derived steroidal derivative produced by metabolism of a plant sterol through specific strains in the additive of the livestock and poultry feed, and based on the molecular structure distribution of the plant-derived steroidal derivative, the hydrophilicity and hydrophobicity of the plant-derived steroidal derivative can be reasonably distributed, so that the stability of the surface activity of the plant-derived steroidal derivative is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of feed additives, and particularly relates to an application of a plant-derived sterol derivative and a feed additive. BACKGROUND

[0002] Feed is widely used in the breeding of livestock, poultry and aquatic animals, and the addition of an appropriate amount of feed additives in feed can meet the nutritional needs of various animals at various growth stages. Energy of feed additives is an important nutritional component of livestock and poultry feed. However, at the present stage, energy feed mainly has two defects: 1. Resource shortage. There is an imbalance between demand and supply of energy feed, and the demand is large while the supply is small, so that the energy feed is in short supply; 2. Low utilization rate of energy feed. Under different production purposes, the utilization efficiency of feed energy by livestock and poultry is quite different; at the present stage, research shows that the conversion efficiency of energy feed from high to low is: maintenance > milk production > growth and fattening > pregnancy or hair production. For example, research at the present stage shows that the conversion efficiency of feed for the growth and fattening of ruminants is 40% to 60%, and the conversion efficiency of feed for pregnancy synthesis is 10% to 30%. In addition, the conversion rate of energy feed is quite different for different animals under different energy components.

[0003] Phytosterol, also known as phytosterol, is a plant sterol compound. The main components of phytosterol include sitosterol, campesterol, stigmasterol, brassicasterol and corresponding alkanols, all of which have a skeleton structure of cyclopentane perhydrophenanthrene as the core. Phytosterol generally contains an alcohol group. The structure of these phytosterols has some side chains at C24 compared to the structure of cholesterols. For example, compared to the structure of cholesterols, sitosterol has an ethyl group at C24, and campesterol has a methyl group at C24, and stigmasterol has an ethyl group at C24 in addition to a double bond at C22. Based on the chemical properties of the above phytosterols, phytosterols exhibit hydrophobicity at the physical level, but some phytosterols have a hydroxyl group in their structure, which makes phytosterols have hydrophilicity. Phytosterols with hydrophilic and hydrophobic properties can effectively reduce the surface tension between oil and water, which indicates that phytosterols may have strong surface activity.

[0004] However, the balance between the hydrophilicity and hydrophobicity of the structure of phytosterols at the present stage is poor, which makes the surface activity of phytosterols unstable. SUMMARY

[0005] The present application provides an application of a plant-derived sterol derivative and a feed additive to solve the technical problem of how to improve the stability of the surface activity of phytosterols.

[0006] In a first aspect, embodiments of the present application provide an application of a plant-derived steroidal derivative. The plant-derived steroidal derivative having a molecular structure as shown in Formula 1 is applied in an additive of livestock and poultry feed.

[0007]

[0008] In Formula 1, R1 and R2 groups respectively include a carbon-carbon single bond or a carbon-carbon double bond.

[0009] The R3 group includes oxygen or a hydroxyl group.

[0010] Optionally, in the case that the R3 group is oxygen, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0011]

[0012]

[0013] Optionally, in the case that the R3 group is a hydroxyl group, the R3 group includes a cis-hydroxyl group or a trans-hydroxyl group.

[0014] Optionally, if the hydroxyl group is a cis-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0015]

[0016]

[0017] Optionally, if the hydroxyl group is a trans-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0018]

[0019]

[0020] In a second aspect, embodiments of the present application provide a feed additive. The feed additive includes the plant-derived steroidal derivative used in the application of the first aspect.

[0021] In a third aspect, embodiments of the present application provide a compound livestock and poultry feed. The compound livestock and poultry feed includes the feed additive of the second aspect.

[0022] Optionally, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.02-10.00):100.

[0023] Optionally, if the compound livestock and poultry feed is used for poultry egg production, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.1-1.0):100.

[0024] Optionally, if the compound livestock and poultry feed is used for poultry or fish weight gain, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.1-10.0):100.

[0025] The above technical solutions provided by the embodiments of the present application have the following advantages compared with the prior art:

[0026] The application of the plant-derived steroidal derivative provided by the embodiments of the present application uses the plant-derived steroidal derivative produced by metabolizing phytosterol by a specific strain as an additive in livestock and poultry feed. Based on the steroidal nucleus and a large number of carboxyl groups of the plant-derived steroidal derivative, the hydrophilicity of the carboxyl group and the lipophilicity of the steroidal nucleus, the plant-derived steroidal derivative has strong surface activity. In addition, the carboxyl groups are distributed at the end of the main chain of the steroidal nucleus. Through the intermolecular force between the carboxyl group and the steroidal nucleus, the hydrophilic group and the steroidal nucleus of the plant-derived steroidal derivative can be uniformly distributed in the molecular structure of the plant-derived steroidal derivative. At the same time, under the action of intermolecular force and solution system, the hydrophilicity and hydrophobicity of the plant-derived steroidal derivative can be reasonably distributed, so as to improve the stability of the surface activity of the plant-derived steroidal derivative. BRIEF DESCRIPTION OF DRAWINGS

[0027] The drawings incorporated into the specification and forming a part thereof, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the application.

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced as follows. Obviously, for those skilled in the art, other drawings can also be obtained from these drawings without creative labor.

[0029] Figure 1 The nuclear magnetic resonance hydrogen spectrum result graph of the plant-derived steroidal derivative in the application of the plant-derived steroidal derivative provided by the embodiments of the present application;

[0030] Figure 2 For Figure 1 The enlarged schematic view of A-A in the middle;

[0031] Figure 3A result chart of liquid chromatography and mass spectrometer combined use of a plant-derived steroidal derivative in the application embodiment of the application provides a plant-derived steroidal derivative; wherein, Figure 3 A is a liquid chromatogram of a plant-derived steroidal derivative, Figure 3 B is a mass spectrum of a plant-derived steroidal derivative. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical scheme and advantages of the embodiments of the present application clearer, the technical scheme in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0033] Various embodiments of the present application can exist in the form of a range; it should be understood that the description in the form of a range is only for the convenience and brevity, and should not be understood as a hard limit on the scope of the present application; therefore, it should be considered that the described range has disclosed all possible sub-ranges and single values in the range; for example, it should be considered that the range from 1 to 6 has disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers in the range, such as 1, 2, 3, 4, 5 and 6, regardless of the range; in addition, whenever a numerical range is indicated in this text, it refers to any cited number (fraction or integer) in the indicated range.

[0034] In this document, the terms "comprise", "comprising", "include", "including" or "contains", "containing" when used mean "including, but not limited to". The terms "first", "second", "third", etc., when used in this document, are used only to distinguish one entity or action from another, and do not necessarily require or imply any actual relationship or order between or among such entities or actions. The term "and / or", describing the association relationship of associated objects, means that there can be three relationships, for example, A and / or B can represent: the case of A alone, the case of A and B existing at the same time, and the case of B alone; wherein A and B can be singular or plural. "At least one" means one or more, and "multiple" means two or more; "at least one", "at least one of the following" or the like means any combination of the items, including single item or combination of multiple items; for example, "at least one of a, b, or c", or "at least one of a, b, and c", can represent a, b, c, a-b (i.e. a and b), a-c, b-c, or a-b-c, wherein a, b, and c can be singular or plural. The "parts ratio" such as weight parts, mass parts, etc. represents the proportional relationship between components. In the proportional relationship described in this document, the parameters that need to be described by the proportion should be understood as the front item of the proportional form according to the order of description, and the proportional number should be understood as the rear item of the proportional form, for example, the mass ratio of substance A, substance B and substance C is 1:2:3, then substance A, substance B and substance C should be one-to-one corresponding in the proportional form according to the description order, i.e. the mass of substance A: the mass of substance B: the mass of substance C = 1:2:3.

[0035] Unless otherwise specifically stated, the various raw materials, reagents, instruments and equipment, etc. used in this document can be purchased on the market or can be prepared by existing methods.

[0036] The plant-derived steroidal derivative provided by the embodiments of the present application is applied in the additive of livestock and poultry feed, and has the molecular structure as shown in formula 1.

[0037]

[0038] In formula 1, the R1 and R2 groups respectively include a carbon-carbon single bond or a carbon-carbon double bond.

[0039] The R3 group includes oxygen or a hydroxyl group.

[0040] It should be noted that the hydroxyl group generally involves cis structure or trans structure.

[0041] It should be noted that the application provided by the embodiment of the present application is a plant-derived sterol derivative, which is related to the biological transformation of phytosterol through microbial metabolic engineering, and the development of a new type of multifunctional feed additive based on its unique molecular structure. Specifically:

[0042] (1) The specific strain (such as M. neoaurum CCTCC AB2019054 strain after specific gene modification) is used to perform directional metabolism on phytosterol, and through biological transformation reactions such as hydroxylation and oxidative dehydrogenation of microbial enzyme system, a sterol derivative with a new type of substituent is generated. The core feature of this derivative is that its molecular structure contains a complete four-ring sterol nucleus and multiple carboxylic acid groups, in which the carboxyl group is covalently connected to the end of the C17 side chain of the sterol nucleus in β-configuration, forming an amphiphilic molecular structure.

[0043] (2) From the perspective of molecular mechanism, the plant-derived sterol derivative will exhibit significant amphiphilic properties: the rigid four-ring structure of the sterol nucleus endows it with excellent liposolubility, which can effectively penetrate the phospholipid bilayer of the cell membrane; and the dense distribution of carboxylic acid groups at the end of the side chain forms a strong hydrophilic region through ionization. It is particularly noteworthy that the spatial arrangement of the carboxyl group adopts a star-shaped topological configuration, and this three-dimensional arrangement can form a stable micellar structure in solution through intermolecular hydrogen bond network and van der Waals force, with a critical micelle concentration (CMC) reduced by about 40% compared to traditional surfactants. Molecular dynamics simulation shows that the orientation angle of the derivative at the oil-water interface can reach 65°±3°, which is significantly higher than that of conventional bile salt surfactants, which makes it have unique advantages in feed lipid emulsification and nutrient component microencapsulation.

[0044] (3) The technical advantages in the application of feed additives are reflected in three dimensions: first, as a biological surfactant, the plant-derived sterol derivative has strong interfacial tension reducing ability; second, the chelation of carboxylic acid groups with metal ions can effectively block the bioavailability of heavy metals in feed; third, the similarity of the sterol nucleus structure to cholesterol in the animal body allows it to competitively inhibit the absorption of exogenous cholesterol in the intestine.

[0045] Therefore, the application provided by the embodiment of the present application is a plant-derived sterol derivative, which breaks through the environmental residue problem of traditional chemical synthesis surfactants and realizes precise modification of molecular structure through biological transformation process. The plant-derived sterol derivative can maintain structural stability in the pH range of 2.5-8.0, and the activity retention rate after resistance to granulation temperature (85℃) treatment is >98%, fully meeting the requirements of feed processing technology. This green manufacturing path based on microbial metabolic engineering provides an innovative solution for the development of new functional feed additives.

[0046] In some alternative embodiments, in the case that the R3 group is oxygen, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0047]

[0048]

[0049] In these embodiments, in the case that the R3 group is oxygen, by matching the R1 and R2 groups of different carbon-carbon single bonds and carbon-carbon double bonds, the plant-derived steroidal derivative can encompass various stable groups (such as 3-carbonyl and carbon-carbon double bond), which have the following properties of the plant-derived steroidal derivative:

[0050] 1. Synergistic effect of 3-carbonyl:

[0051] (1) Enhancing molecular structure stability:

[0052] The introduction of 3-carbonyl allows the carbon ring structure of the steroidal nucleus to form a conjugated system, which reduces the intramolecular energy of the plant-derived steroidal derivative through electron delocalization effect, thereby improving the thermal stability of the plant-derived steroidal derivative.

[0053] (2) Optimizing bioactive sites: 3-carbonyl can form specific binding with intestinal cholesterol absorption protein (NPC1L1) as a hydrogen bond acceptor, competitively inhibiting the efficiency of cholesterol absorption. At the same time, the carbon site of the carbonyl group can bind to the active center of microbial metabolic enzymes (such as CYP450), promoting the carboxylation reaction of the side chain of the plant-derived steroidal derivative, thereby improving the performance stability of the plant-derived steroidal derivative.

[0054] 2. Functional expansion of carbon-carbon double bond:

[0055] (1) Constructing reactive centers: The π-electron cloud of the carbon-carbon double bond can be used as a nucleophilic reagent attack site, which can be catalyzed by related biological enzymes in the animal body or inorganic catalysts outside the body, etc. to realize directional modification of molecules. These modified plant-derived steroidal derivatives can greatly improve the hydrophilicity or hydrophobicity, thereby meeting the different characteristic needs of the plant-derived steroidal derivative.

[0056] (2) Regulating the balance of amphiphilicity: The cis / trans isomerism of the carbon-carbon double bond can affect the steroidal nucleus conformation of the plant-derived steroidal derivative: the cis configuration reduces the steroidal nucleus plane angle by 12°, promoting the wedge-shaped insertion of the carboxyl side chain into the lipid bilayer, which can effectively reduce the interfacial tension of the plant-derived steroidal derivative. The trans configuration can enhance the rigidity of the molecule to reduce the critical micelle concentration of the plant-derived steroidal derivative.

[0057] 3. Synergistic mechanism

[0058] (1) Electron effect transmission: the 3-carbonyl group can activate the adjacent double bond of the plant-derived steroidal derivative through conjugation effect, so as to reduce the oxidation potential of the plant-derived steroidal derivative, which makes the plant-derived steroidal derivative more prone to enzymatic oxidation reaction and generates an α, β-unsaturated ketone structure, which can improve the chelating ability of the plant-derived steroidal derivative to heavy metal ions, effectively promoting the digestion and decomposition of the plant-derived steroidal derivative.

[0059] (2) Spatial conformation regulation:

[0060] The 1,3-conjugated system formed by the 3-carbonyl group and the carbon-carbon double bond can cause the chair-boat conformational transition of the steroidal nucleus of the plant-derived steroidal derivative, so as to expose the carboxyl group of the side chain of the plant-derived steroidal derivative, which can improve the binding efficiency of the plant-derived steroidal derivative to calcium ions in livestock and poultry feed, thereby effectively reducing the loss of mineral elements in livestock and poultry feed.

[0061] 4. Application performance improvement:

[0062] (1) Nutrient carrier function: the epoxy structure generated by the oxidation of the carbon-carbon double bond can be used as a molecular anchoring site for vitamin E, improving the encapsulation rate of fat-soluble nutrients and prolonging the slow-release time of fat-soluble nutrients, which is beneficial to the digestion and absorption of livestock and poultry animals to livestock and poultry feed.

[0063] (2) Metabolic regulation: the 3-carbonyl group and the steroidal nucleus of the plant-derived steroidal derivative can mimic the brassinosteroid signal molecule, activate the PPAR-γ pathway of livestock and poultry, and down-regulate the expression of genes related to adipocyte differentiation in livestock and poultry animals, thereby improving the muscle protein synthesis rate of livestock and poultry animals, to promote the digestion and absorption of livestock and poultry animals to livestock and poultry feed.

[0064] In some optional embodiments, when the R3 group is a hydroxyl group, the R3 group includes a cis-hydroxyl group or a trans-hydroxyl group.

[0065] In some optional embodiments, if the hydroxyl group is a cis-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0066]

[0067] In some optional embodiments, if the hydroxyl group is a trans-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0068]

[0069] In these embodiments, for the case of R3group in cis-hydroxyl or trans-hydroxyl, by matching different R1and R2groups of carbon-carbon single bond and carbon-carbon double bond, the phytosteroid derivative can cover a variety of functional groups (such as hydroxyl, 3-carbonyl and carbon-carbon double bond), which have the following characteristics:

[0070] 1. Functional gain of hydroxyl introduction:

[0071] (1) Enhanced water solubility and bioavailability:

[0072] The polar nature of the hydroxyl group can significantly improve the water solubility of the phytosteroid derivative, especially after introducing a hydroxyl group at the carbon site of the phytosteroid derivative, the water solubility can be improved by up to 4.6 times or more. This feature is conducive to the dispersion of the phytosteroid derivative in the animal digestive tract and the release of active ingredients.

[0073] (2) Strengthening intermolecular forces:

[0074] As a hydrogen bond donor / acceptor, the hydroxyl group can form stable binding with intestinal cholesterol transport proteins (such as NPC1L1) to inhibit the absorption efficiency of cholesterol. At the same time, the hydroxyl group interacts with the polar head group of the lipid bilayer to promote the formation of the emulsion system of the phytosteroid derivative.

[0075] (3) Activating antioxidant and anti-inflammatory pathways:

[0076] The hydroxyl group plays an antioxidant role by scavenging free radicals and reduces the expression of inflammatory factors (such as IL-6, TNF-α) in livestock and poultry animals by inhibiting the NF-κB signaling pathway, to increase the feed intake of livestock and poultry animals.

[0077] 2. Structural optimization of 3-carbonyl introduction:

[0078] (1) Stable molecular conformation:

[0079] The 3-carbonyl group can form a conjugated system with the steroidal ring nucleus of the phytosteroid derivative, reducing the intramolecular energy of the phytosteroid derivative through electron delocalization effect, and promoting the phytosteroid derivative to have higher thermal stability during feed processing.

[0080] (2) Metabolic targeting:

[0081] The 3-carbonyl group can mimic the structural features of endogenous steroid hormones (such as cortisol), selectively activate PPARγ nuclear receptors, promote the expression of fat metabolism-related genes in livestock and poultry animals, and reduce serum LDL levels, promoting digestion and absorption in livestock and poultry animals.

[0082] (3) Enhanced interfacial activity:

[0083] 3-carbonyl can form an amphiphilic structure with the side chain carboxyl of the plant-derived steroidal derivative, so that the critical micelle concentration (CMC) of the plant-derived steroidal derivative is reduced by more than 40% compared with that of natural sterol, and the oil-water interfacial tension of the plant-derived steroidal derivative is also reduced, thereby significantly improving the encapsulation rate of fat-soluble nutrients (such as vitamin A) in livestock and poultry animals.

[0084] 3. Synergistic mechanism of hydroxyl and 3-carbonyl:

[0085] (1) Electron effect transmission:

[0086] The electron-donating effect of the hydroxyl group is transmitted to the 3-carbonyl group through the steroidal ring nucleus, reducing the redox potential of the 3-carbonyl group by 0.25 V and making the 3-carbonyl group more easily participate in enzymatic reactions to generate an α-keto acid intermediate, thereby enhancing the heavy metal ion chelating ability of the plant-derived steroidal derivative.

[0087] (2) Spatial conformation complementarity:

[0088] The intramolecular hydrogen bond network formed by the hydroxyl group and the 3-carbonyl group can induce a chair-boat conformational transition of the steroidal ring nucleus, thereby exposing an alternating arrangement region of hydrophobic regions and hydrophilic groups, which can improve the cell membrane penetration efficiency of the plant-derived steroidal derivative.

[0089] (3) Metabolic transformation synergy:

[0090] The 3-carbonyl group serves as an activation site for microbial metabolism and can be reduced to a hydroxyl group under the action of intestinal flora to form a dynamic redox cycle system, which can prolong the half-life of the active ingredients of the plant-derived steroidal derivative.

[0091] Based on a general inventive concept, the embodiments of the present application provide a feed additive, which comprises the plant-derived steroidal derivative used in the application.

[0092] The feed additive is realized based on the above-mentioned application, and the specific principles of the application can refer to the above-mentioned embodiments. Since the feed additive adopts part or all of the technical solutions of the above-mentioned embodiments, it at least has all the beneficial effects brought by the technical solutions of the above-mentioned embodiments, which will not be repeated here.

[0093] It should be noted that the plant-derived steroidal derivative has high surface activity, which enables the plant-derived steroidal derivative to effectively promote fat emulsification and form fat acid chylomicrons that can be suspended in a hydrophilic system. These fat acid chylomicrons can fully contact lipase in the body of livestock and poultry animals, so that the fat acid chylomicrons are decomposed under the action of lipase to accelerate the digestion and absorption of fat in the body of livestock and poultry animals.

[0094] Based on one general inventive concept, the embodiments of the present application provide a compound livestock and poultry feed, which comprises the feed additive.

[0095] The compound livestock and poultry feed is realized based on the above-mentioned feed additive, and the specific composition of the feed additive can refer to the above-mentioned embodiments. Since the compound livestock and poultry feed adopts part or all of the technical solutions of the above-mentioned embodiments, it at least has all the beneficial effects brought by the technical solutions of the above-mentioned embodiments, which will not be repeated here.

[0096] In some optional embodiments, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.02-10.00):100.

[0097] In these embodiments, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed can satisfy the relationship: m1:m2=(0.02-10.00):100, so that the compound livestock and poultry feed has sufficient feed additive, and the sufficient feed additive has good surface activity, which can promote the emulsification of the fat of the plant-derived steroidal derivative through the feed additive, thereby improving the digestion and absorption efficiency of the fat in the livestock and poultry animals.

[0098] The mass m1 of the feed additive can be 0.02, 0.03, 0.04, 0.05, 0.10, 0.20, 0.30, 0.40, 0.50, 1.00, 2.00, 3.00, 4.00, 5.00, 6.00, 7.00, 8.00, 9.00 or 10.00.

[0099] In some optional embodiments, if the compound livestock and poultry feed is used for egg production of poultry, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.1-1.0):100.

[0100] In these embodiments, if the compound livestock and poultry feed is used for egg production of poultry, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed can satisfy the relationship: m1:m2=(0.1-1.0):100, so that the compound livestock and poultry feed has sufficient feed additive, and the sufficient feed additive has good surface activity, which can promote the emulsification of the fat of the plant-derived steroidal derivative through the feed additive, thereby improving the digestion and absorption efficiency of the fat in the livestock and poultry animals.

[0101] The mass m1 of the feed additive can be 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1.00.

[0102] In some optional embodiments, if the compound livestock and poultry feed is used for weight gain of poultry or fish, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the relationship: m1:m2=(0.1-10.0):100.

[0103] In these embodiments, if the compound livestock and poultry feed is used for weight gain of poultry or fish, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed can satisfy the relationship: m1:m2=(0.1-10.0):100, so that the compound livestock and poultry feed has sufficient feed additives, and the sufficient feed additives have good surface activity, and the fats of the plant-derived steroidal derivatives can be emulsified by the feed additives, thereby improving the digestion and absorption efficiency of the fats in the livestock and poultry animals.

[0104] The mass m1 of the feed additive can be 0.1, 0.2, 0.3, 0.4, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 or 10.0.

[0105] The present application will be further described below in conjunction with specific examples. The experimental methods in the following examples are generally determined according to national standards / industry standards, unless otherwise specified. If there is no corresponding national standard / industry standard, the general international standard, conventional conditions or the conditions recommended by the manufacturer are used.

[0106] Example 1

[0107] The metabolic products of existing 3-OPCM and 3-OPDCM are structurally modified to obtain plant-derived steroidal derivatives with the molecular structure of Formula 1:

[0108]

[0109] According to different modification sites and modification groups, the plant-derived steroidal derivatives can be divided into two categories containing oxygen and containing hydroxyl groups, specifically:

[0110] In the case of R3 group being oxygen, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0111]

[0112]

[0113] If the R3 group is a cis-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0114]

[0115]

[0116] If the R3 group is a trans-hydroxyl group, the molecular structure of the plant-derived steroidal derivative includes at least one of the following molecular structures:

[0117]

[0118]

[0119] In the molecular structure of these plant-derived steroidal derivatives, Formula 2 is the carboxylic acid structure formed after the ester bond of 3-OPDCM is hydrolyzed, and the specific flow is as follows:

[0120] 30 g of 3-OPDCM is added to 120 g of a NaOH solution with a mass concentration of 20%, to obtain a mixture, the mixture is warmed to 40-50°C and reacted under stirring for 3 h, at which time thin layer chromatography (TLC, using a volume ratio of petroleum ether to ethyl acetate of 2:1) shows that 3-OPDCM disappears, to obtain a reaction product; the reaction product is cooled to 10-15°C, 3 mol / L hydrochloric acid is slowly added dropwise to adjust the pH to 1-2, then suction filtration is performed, and the filter cake is washed with water until the pH is 6-7, the filter cake after washing is placed in a blast drying oven at 50°C for 16 h, and the collected material is a white solid, which is a compound of Formula 2, and the product yield is 92%.

[0121] Formula 3 is the carboxylic acid structure formed after the ester bond of 3-OPCM is hydrolyzed, and the specific flow is as follows:

[0122] 30 g of 3-OPDCM is added to 120 g of a NaOH solution with a mass concentration of 20%, to obtain a mixture, the mixture is warmed to 40-50°C and reacted under stirring for 3 h, at which time thin layer chromatography (TLC, using a volume ratio of petroleum ether to ethyl acetate of 2:1) shows that 3-OPDCM disappears, to obtain a reaction product; the reaction product is cooled to 10-15°C, 3 mol / L hydrochloric acid is slowly added dropwise to adjust the pH to 1-2, then suction filtration is performed, and the filter cake is washed with water until the pH is 6-7, the filter cake after washing is placed in a blast drying oven at 50°C for 16 h, and the collected material is a white solid, which is a compound of Formula 2, and the product yield is 93%.

[0123] Formula 4 is obtained based on the reduction of the double bond at the α position of the 3-carbonyl group in the molecular structure of Formula 2, and the specific flow is as follows:

[0124] In an autoclave, 5 g of the plant-derived steroidal derivative of Formula 2 was added to 50 mL of methanol, and 0.05 g of Pd / C catalyst with a mass concentration of 10% was added. The reaction was carried out by replacing the atmosphere with nitrogen three times, replacing it with hydrogen three times, and pressurizing to 0.1 MPa for 1 h. After the reaction was completed, the reaction product was suction filtered, and the filter cake was washed with 20 mL of methanol. The filtrate was concentrated to dryness at 50°C to obtain 4.8 g of a white solid, which was the plant-derived steroidal derivative of Formula 4, with a product yield of 96%.

[0125] Similarly, Formula 5 was obtained by reducing the double bond at the α position of the 3-carbonyl group in the molecular structure of Formula 3. The specific process is as follows:

[0126] In an autoclave, 5 g of the plant-derived steroidal derivative of Formula 3 was added to 50 mL of methanol, and 0.05 g of Pd / C catalyst with a mass concentration of 10% was added. The reaction was carried out by replacing the atmosphere with nitrogen three times, replacing it with hydrogen three times, and pressurizing to 0.1 MPa for 1 h. After the reaction was completed, the reaction product was suction filtered, and the filter cake was washed with 20 mL of methanol. The filtrate was concentrated to dryness at 50°C to obtain 4.6 g of a white solid, which was the plant-derived steroidal derivative of Formula 5, with a product yield of 91%.

[0127] Formula 6 is a cis-hydroxyl group obtained by reducing the 3-carbonyl group in the molecular structure of Formula 2, and Formula 10 is a trans-hydroxyl group obtained by reducing the 3-carbonyl group in the molecular structure of Formula 2. The specific process is as follows:

[0128] In an autoclave, 5 g of the plant-derived steroidal derivative of Formula 2 was added to 50 mL of methanol, and 0.05 g of Pd / C catalyst with a mass concentration of 10% was added. The reaction was carried out by replacing the atmosphere with nitrogen three times, replacing it with hydrogen three times, and pressurizing to 0.1 MPa for 1 h. After the reaction was completed, the reaction product was suction filtered, and the filter cake was washed with 20 mL of methanol. The filtrate was concentrated to dryness at 50°C to obtain 4.8 g of a white solid, which was the plant-derived steroidal derivative of Formula 4, with a product yield of 96%.

[0129] Similarly, Formula 7 is a cis-hydroxyl group obtained by reducing the 3-carbonyl group in the molecular structure of Formula 3, and Formula 11 is a trans-hydroxyl group obtained by reducing the 3-carbonyl group in the molecular structure of Formula 3. The specific process is as follows:

[0130] Take 2.34 g of NaOH into 20 mL of water, add 5 g of plant sterol derivative of formula 3, stir until the solid is completely dissolved, then slowly add 2.25 g of NaBH4, react overnight at room temperature, after the reaction is complete, adjust the pH to 1-2 with 4 mol / L hydrochloric acid, filter, and obtain a mixture of plant sterol derivative of formula 7 and plant sterol derivative of formula 11, which are separated by silica gel column chromatography to obtain 1.3 g of plant sterol derivative of formula 7 and 1.2 g of plant sterol derivative of formula 11, both of which are white solids.

[0131] Similarly, formula 8 is a cis-hydroxyl group obtained by reduction of the 3-carbonyl group in the molecular structure of formula 4, and formula 12 is a trans-hydroxyl group obtained by reduction of the 3-carbonyl group in the molecular structure of formula 4, and the specific process is as follows:

[0132] Take 2.34 g of NaOH into 20 mL of water, add 5 g of plant sterol derivative of formula 4, stir until the solid is completely dissolved, then slowly add 2.25 g of NaBH4, react overnight at room temperature, after the reaction is complete, adjust the pH to 1-2 with 4 mol / L hydrochloric acid, filter, and obtain a mixture of plant sterol derivative of formula 8 and plant sterol derivative of formula 12, which are separated by silica gel column chromatography to obtain 1.1 g of plant sterol derivative of formula 8 and 1.0 g of plant sterol derivative of formula 12, both of which are white solids.

[0133] Similarly, formula 9 is a cis-hydroxyl group obtained by reduction of the 3-carbonyl group in the molecular structure of formula 5, and formula 13 is a trans-hydroxyl group obtained by reduction of the 3-carbonyl group in the molecular structure of formula 5, and the specific process is as follows:

[0134] Take 2.34 g of NaOH into 20 mL of water, add 5 g of plant sterol derivative of formula 5, stir until the solid is completely dissolved, then slowly add 2.25 g of NaBH4, react overnight at room temperature, after the reaction is complete, adjust the pH to 1-2 with 4 mol / L hydrochloric acid, filter, and obtain a mixture of plant sterol derivative of formula 9 and plant sterol derivative of formula 13, which are separated by silica gel column chromatography to obtain 1.4 g of plant sterol derivative of formula 9 and 1.3 g of plant sterol derivative of formula 13, both of which are white solids.

[0135] Example 2

[0136] The plant-derived steroidal derivatives shown in Formula 2 to Formula 13 disclosed in Example 1 are respectively added into ordinary hen laying feed as feed additives in different addition amounts (0.1%, 0.3%, 0.6%, 1.0%), mixed to obtain various laying livestock and poultry feeds. Among them, the ordinary hen laying feed uses egg hen laying period compound feed 524 produced by Shijiazhuang Zhengda Co., Ltd.; the main components of the ordinary hen laying feed include, in mass fraction: crude protein ≥ 16.5%, crude fiber ≤ 7.0%, crude ash ≤ 15.0, calcium: 2.80%~4.20%, total phosphorus ≥ 0.50%, sodium chloride 0.2%~0.8%, total amount of methionine + cystine ≥ 0.65%, and water content ≤ 14.0%.

[0137] Example 3

[0138] Corn, soybean meal and wheat bran are used as main raw materials to prepare a basic daily ration, and the specific composition of the basic daily ration includes, in mass fraction: corn: 56 parts~59 parts, soybean meal: 32 parts~35 parts, and wheat bran: 6 parts~12 parts.

[0139] The plant-derived steroidal derivatives shown in Formula 2 to Formula 13 disclosed in Example 1 are respectively added into the basic daily ration as feed additives in different addition amounts (0.2%, 0.5%, 2.0%, 5.0%, 10.0%), mixed to obtain weight gain livestock and poultry feed.

[0140] Example 4

[0141] The plant-derived steroidal derivatives shown in Formula 2 to Formula 13 disclosed in Example 1 are respectively added into ice fresh feed as feed additives in an addition amount of 0.1%, mixed to obtain aquatic livestock and poultry feed. The ice fresh feed uses Hailin brand large yellow croaker compound feed (floating special purpose) produced by Xiamen Hailin Biological Technology Co., Ltd.; the main components of the ice fresh feed include, in mass fraction: crude protein ≥ 45.0%, crude fat ≥ 5.0%, crude fiber ≤ 5.0%, crude ash ≤ 15.0%, lysine ≥ 2.4%, calcium: 1.4%~4.5%, total phosphorus: 1.0%~2.2%, and water content ≤ 11.0%.

[0142] Related experiments and effect data:

[0143] 1. The plant-derived steroidal derivative shown in Formula 2 disclosed in Example 1 is actually measured, and the nuclear magnetic resonance hydrogen spectrum is shown in Figure 1 and Figure 2 The results show that the molecular formula of the plant-derived steroidal derivative is C 22 H 30 O3, the plant-derived steroidal derivative shown in Formula 2 contains 30 hydrogen atoms, and the nuclear magnetic resonance hydrogen spectrum result is:

[0144] 1 HNMR(400MHz,DMSO-d6): δ11.97(s, 1 H), 5.64(d, 0.5 H), 2.70-2.77(q, 1 H), 2.57(m, 1 H), 2.15-2.41(m, 4.5 H), 1.97(m, 1 H), 1.88-2.00 (m, 4 H), 1.50-1.77 (m, 6 H),

[0145] 1.13-1.36(m, 6 H), 0.70-1.06(m, 5 H).

[0146] The molecular weight M of the plant-derived steroid derivative shown in Formula 2 is 342.22. The plant-derived steroid derivative was detected by liquid chromatography-mass spectrometry (LC-MS), and the results are as follows: Figure 3 As shown. By Figure 3 A and Figure 3 As can be seen from B, the plant-derived steroid derivative shown in Formula 2 has a molecular ion peak of 343.2, which is the M+1 peak, further confirming the chemical structure of the plant-derived steroid derivative shown in Formula 2.

[0147] 2. Actual feeding experiments were conducted on the egg-laying livestock and poultry feeds of Example 2. The experimental procedure for each egg-laying livestock and poultry feed was as follows:

[0148] One hundred hens of the same age (25 weeks) at their peak egg production were selected and randomly divided into one control group and four experimental groups, with 20 hens in each group. Under the same feeding conditions such as light, temperature, humidity, and sufficient water, the control group was fed ordinary egg-laying feed, while the experimental groups were fed different amounts of plant-derived steroidal derivatives with molecular structures shown in Formulas 2 to 13.

[0149] During the 100-day egg-laying cycle, the number of eggs laid by each group of 20 hens was counted, as shown in Table 1.

[0150] Table 1. Distribution of egg production by hen in each group.

[0151]

[0152]

[0153]

[0154]

[0155] 3. The weight gain livestock feed of Example 3 is subjected to actual feeding experiments, and the experimental procedure for each weight gain livestock feed is as follows:

[0156] 480 one-day-old fast-growing white-feathered broilers with similar body weights are selected and randomly divided into 6 groups, with 4 replicates in each group and 20 in each replicate. The whole test period is 42 days. The control group is only fed with the basic diet, and the experimental groups are added with plant-derived sterol derivatives with different molecular structures shown in Formula 2 to Formula 13 on the basis of the basic diet. The body weight changes of the white-feathered broilers during the test period are counted, and the results are shown in Table 2.

[0157] Table 2: Body weight changes of white-feathered broilers in each group

[0158]

[0159]

[0160]

[0161]

[0162]

[0163] 4. The aquaculture livestock feed of Example 4 is subjected to actual feeding experiments, and the experimental procedure for each aquaculture livestock feed is as follows:

[0164] About 26000 large yellow croakers with an initial body weight of about 120g are selected and randomly divided into a control group and multiple experimental groups, with about 2000 in each group. Then the large yellow croakers are raised in net cages. The control group is only fed with ice fresh feed, and the experimental groups are added with plant-derived sterol derivatives with different molecular structures shown in Formula 2 to Formula 13 on the basis of the ice fresh feed according to an addition amount of 0.1%. After 28 days of feeding, the growth state changes of the large yellow croakers during the test period are counted, and the results are shown in Table 3.

[0165] Table 3: Growth state changes of large yellow croakers in each group

[0166]

[0167]

[0168] As shown in Tables 1 to 3, the application provides the application of the plant-derived sterol derivative. The plant-derived sterol derivative produced by the metabolism of the specific strain of the plant sterol is used as an additive in the livestock and poultry feed. Based on the molecular structure distribution of the plant-derived sterol derivative, the hydrophilicity and hydrophobicity of the plant-derived sterol derivative can be reasonably distributed to improve the stability of the surface activity of the plant-derived sterol derivative. The feed added with the plant-derived sterol derivative can increase the daily egg production of the hens by 1 to 5 eggs in a 100-day egg production period. In the 42-day weight gain test of the basic diet, the body weight of the white-feathered broiler chickens can be increased by more than 30 g. In the 28-day aquaculture test, the growth state of the large yellow croaker can be obviously improved.

[0169] In addition, the application provides a compound livestock and poultry feed. The compound livestock and poultry feed uses the plant-derived sterol derivative as a feed additive. The high stability of the surface activity of the plant-derived sterol derivative can effectively promote the fat emulsification of the livestock and poultry feed to form fat acid chylomicron particles that can be suspended in the hydrophilic system. The fat acid chylomicron particles can fully contact with lipase in the livestock and poultry animal body, so that the fat acid chylomicron particles are decomposed under the action of the lipase to accelerate the digestion and absorption of fat in the livestock and poultry animal body.

[0170] In addition, the application provides a compound livestock and poultry feed. The compound livestock and poultry feed is found through the actual application experiment that it can significantly improve the egg production rate of poultry, promote the growth and weight gain of livestock and poultry, improve the meat production performance, and reduce animal malaria and the like when the compound livestock and poultry feed is added into ordinary livestock and poultry feed, basic diet and ice fresh feed.

[0171] The above description is only the specific implementation of the application, which enables those skilled in the art to understand or implement the application. Various modifications of these embodiments will be apparent to those skilled in the art, and the general principles defined in the application can be implemented in other embodiments without departing from the spirit or scope of the application. Therefore, the application will not be limited to these embodiments shown in the application, but will conform to the widest scope consistent with the principles and novel features of the application.

Claims

1. The application of a plant-derived steroid derivative in the preparation of an additive for livestock and poultry feed, wherein a plant-derived steroid derivative having a molecular structure as shown in Formula 1 is applied to an additive for livestock and poultry feed. ， Formula 1, In Formula 1, the R1 and R2 groups respectively include carbon-carbon single bonds or carbon-carbon double bonds; The R3 group includes oxygen or hydroxyl groups; When the R3 group is oxygen, the molecular structure of the plant-derived steroid derivative includes at least one of the following molecular structures: ， Equation 2, ， Formula 3, ， Equation 4 and ， Formula 5; When the R3 group is a hydroxyl group, the R3 group includes cis-hydroxyl or trans-hydroxyl groups; If the hydroxyl group is a cis-hydroxyl group, then the molecular structure of the plant-derived steroid derivative includes at least one of the following molecular structures: ， Formula 6, ， Equation 7, ， Formula 8 and ， Equation 9; If the hydroxyl group is a trans-hydroxyl group, then the molecular structure of the plant-derived steroid derivative includes at least one of the following molecular structures: ， Formula 10, ， Formula 11, ， Equation 12 and ， Formula 13.

2. A compound livestock and poultry feed, wherein the compound livestock and poultry feed comprises the plant-derived steroidal derivatives used in the application of claim 1.

3. According to claim 2, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the following relationship: m1:m2 = (0.02~10.00):

100.

4. According to claim 3, if the compound livestock and poultry feed is used for poultry egg production, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the following relationship: m1:m2 = (0.1~1.0):

100.

5. According to claim 4, if the compound livestock and poultry feed is used for weight gain in poultry or fish, the mass m1 of the feed additive and the mass m2 of the compound livestock and poultry feed satisfy the following relationship: m1:m2 = (0.1~10.0):100.

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