A high-protein nutritional liquid composition and method of making the same

Abstract

The application provides a high-protein nutrient liquid composition and a preparation method thereof, and belongs to the technical field of food processing. The technical problem to be solved is that in the prior art, multi-source proteins are prone to demulsification, have poor taste, and are unstable. The high-protein nutrient liquid composition provided by the application comprises the following raw materials: a composite protein, a composite modified peptide, medium-chain triglycerides, sunflower seed oil, fish oil, phytosterol esters, vitamins, minerals, water and auxiliary materials; the composite protein is prepared by enzymatic reaction of pea protein isolate, chickpea protein isolate and whey protein isolate; the composite modified peptide is prepared by fermentation, enzymatic reaction and glycosylation grafting modification of deep-sea fish skin collagen and defatted walnut meal; and the auxiliary materials are selected from at least one of glycine, citric acid, microcrystalline cellulose and low-acyl gellan gum.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology and relates to a high-protein nutrient solution composition and its preparation method. Background Technology

[0002] High-protein nutritional solutions, as a high-density, easily absorbed liquid nutritional supplement, are widely used in special medical purpose formula foods, clinical enteral nutrition support, and sports nutrition. A study titled "The Effect of High-Energy Nutritional Solution Supplementation on Anti-Fatigue in Adolescent Athletes" (Sports World Academic, Li Jingsong) reveals that soybean polypeptide-based high-energy nutritional solutions can indeed improve the anti-fatigue ability of adolescent athletes. To meet comprehensive and balanced nutritional needs, such products typically require the formulation of multiple animal and plant-derived proteins, small-molecule active peptides, complex lipids of different polarities (such as medium- and long-chain lipids and oils rich in polyunsaturated fatty acids), and abundant vitamins and minerals.

[0003] However, the development and production of existing high-protein liquid nutrient solutions generally face the following three major technical bottlenecks due to the extremely complex composition of the system: First, the coexistence of multiple protein sources easily leads to phase separation and precipitation, while the coexistence of multiple polar lipids easily results in demulsification and flocculation. Current technologies typically involve the direct physical mixing of animal and plant protein powders from different sources. Due to the significant differences in isoelectric point, surface hydrophobicity, and molecular spatial conformation among proteins from different sources, thermodynamic incompatibility and interfacial competitive adsorption easily occur in aqueous liquid systems, leading to protein molecule aggregation and precipitation. Simultaneously, the significant differences in polarity among the compound lipids in the formulation make it difficult for traditional small-molecule emulsifiers to form a high-mechanical-strength interfacial film at the complex oil-water interface. After long-term storage, temperature fluctuations, or heat treatment, the product is highly susceptible to demulsification, flocculation, and lipid flocculation, severely affecting the product's appearance uniformity and physical stability.

[0004] Secondly, small-molecule bioactive peptides are prone to developing unpleasant flavors and have poor thermodynamic stability. To improve the bioactivity and intestinal absorption rate of products, small-molecule bioactive peptides derived from the deep enzymatic hydrolysis of animal and plant proteins are often added to the formulation. However, these small-molecule peptides expose a large number of hydrophobic amino acid residues during enzymatic hydrolysis, resulting in a strong bitter taste and unpleasant aftertaste, severely reducing patient compliance. In addition, commercially available over-hydrolyzed small-molecule peptides, due to their small molecular weight and lack of steric hindrance, are highly susceptible to intermolecular thermal aggregation and uncontrollable Maillard browning when subjected to intense heat treatments such as ultra-high temperature (UHT) sterilization. This leads to a darker product color, abnormal viscosity fluctuations, and a significant loss of the peptide's biological potency.

[0005] Third, micronutrients have poor chemical stability, resulting in significant nutrient degradation and deterioration during their shelf life. Fat-soluble vitamins and polyunsaturated fatty acids are extremely sensitive to oxygen, light, and heat, readily undergoing lipid peroxidation and becoming ineffective, producing off-flavors. Simultaneously, free or simple inorganic mineral salts added to the formula not only easily combine with large protein molecules or phytic acid in the system to form insoluble precipitates, but the free metal ions they release also act as potent pro-oxidants, accelerating the oxidative rancidity of lipids in the system. This leads to low vitamin retention rates and a high susceptibility to mineral precipitation and lipid oxidation deterioration during the product's normal shelf life.

[0006] For example, Chinese patent application CN109744317A discloses a high-protein sports milk containing DHA algal oil and its preparation method. The high-protein sports milk containing DHA algal oil includes: 94-97 parts by weight of milk; 2.0-3.2 parts by weight of concentrated milk protein; 0.4-1.6 parts by weight of casein; 0.1-0.4 parts by weight of whey protein; 0.01-0.05 parts by weight of Peptide; 0.05-0.6 parts by weight of DHA algal oil; 0.05-0.1 parts by weight of complex minerals; 0.03-0.05 parts by weight of complex vitamins; 0.05-0.2 parts by weight of stabilizer; and 0.045-0.4 parts by weight of antioxidant. However, this patent does not provide any efficacy verification. Chinese patent application CN108835262A discloses a modified milk powder for improving memory and its preparation method. The modified milk powder comprises the following components: 40-50 parts whole milk powder or an equivalent dry matter whole milk emulsion; 17-25 parts skim milk powder or an equivalent dry matter skim milk emulsion; 15-23 parts whey powder; 5-15 parts glucose syrup solids or lactose; 2-5 parts vegetable oil; 0.2-3 parts fish oil extract; 0.1-1 part hydrolyzed egg yolk powder; 0.01-2 parts food-grade medicinal materials; 0.01-3 parts dietary fiber; 0.2-2 parts complex minerals; 0.01-0.15 parts complex vitamins; 0.01-0.5 parts sialic acid; 0.025-0.6 parts phosphatidylserine; and 0.002-0.008 parts zeaxanthin. However, it does not include tests on taste or stability.

[0007] Therefore, there is an urgent need in the field for a high-protein nutrient solution composition and its preparation method that has high physical stability, excellent sensory flavor and long-lasting nutrient retention rate, in order to overcome the technical defects of existing technologies such as multi-source protein precipitation, peptide bitterness and heat aggregation, and micronutrient attenuation. Summary of the Invention

[0008] The purpose of this invention is to provide: A high-protein nutrient solution composition and its preparation method, as well as related technologies, are disclosed to solve technical problems such as easy demulsification, poor taste, and instability of multi-source proteins, or combinations thereof.

[0009] To achieve the above-mentioned objectives of the invention, In a first aspect, the present invention provides a high-protein nutrient solution composition comprising the following raw materials: a complex protein, a complex modified peptide, medium- and long-chain triglycerides, sunflower seed oil, fish oil, phytosterol esters, vitamins, minerals, water, and excipients; wherein the complex protein is prepared by enzymatic hydrolysis of pea protein isolate, chickpea protein isolate, and whey protein isolate; wherein the complex modified peptide is prepared by fermentation, enzymatic hydrolysis, and glycosylation grafting modification of deep-sea fish skin collagen and defatted walnut meal; and wherein the excipients are selected from at least one of glycine, citric acid, microcrystalline cellulose, and low-acyl gellan gum.

[0010] Preferably, the method for preparing the composite protein includes the following steps: (1) Mix pea protein isolate and chickpea protein isolate with water, add alkaline protease to hydrolyze for 30-40 minutes, inactivate the enzyme, and obtain protein solution; (2) Mix the protein solution with whey protein isolate, add ferulic acid and laccase for enzymatic hydrolysis and enzyme inactivation, and the protein is obtained.

[0011] Preferably, the mass ratio of whey protein isolate, pea protein isolate, and chickpea protein isolate is 3-5:2-4:2-4.

[0012] More preferably, the mass ratio of the whey protein isolate, pea protein isolate, and chickpea protein isolate is 4:3:3.

[0013] More preferably, the method for preparing the composite protein includes the following steps: (1) Mix pea protein isolate and chickpea protein isolate with water, the mass percentage of pea protein isolate and chickpea protein isolate in water is 5-12%, adjust the pH to 7.5-8.0, add alkaline protease and enzymatically hydrolyze at 45-55℃ for 20-40 min, the amount of alkaline protease added is 400-600 U / g, the amount added is based on the total dry matter mass of pea protein isolate and chickpea protein isolate; heat to 85-90℃ to inactivate enzyme for 8-10 min, cool to 35-45℃ to obtain protein solution; (2) The protein solution from step (1) is mixed with whey protein isolate, and ferulic acid and laccase at a mass of 0.5-1% of the total dry matter of protein in the system and 4-8 U / g are added. The amount of laccase added is based on the mass of the total dry matter of protein in the system. The pH is adjusted to 6-6.8, and the mixture is stirred at a constant temperature of 35-40℃ for 2-3 hours. The temperature is then raised to 85-90℃ to inactivate the enzyme for 8-10 minutes. The mixture is then dried to obtain the final product.

[0014] Preferably, the preparation method of the composite modified peptide includes the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal are mixed with water, sterilized, inoculated with Lactobacillus plantarum and Bacillus subtilis, fermented, inactivated, and fermented to obtain fermentation broth; S2: Cool the fermentation broth to 45-55℃, adjust the pH to 8-8.5, add alkaline protease and hydrolyze for 1.5-2.5h; then adjust the pH to 7.0-7.5, add flavor protease and specific leucine aminopeptidase, continue hydrolysis for 2-3h, inactivate the enzyme, and obtain the hydrolysate. S3: After concentrating the enzymatic hydrolysate, add 1.5-2.0 times the dry matter weight of the enzymatic hydrolysate, add galactooligosaccharides (DP3-5), adjust the pH to 7.5-8.0, react at 60-65℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction to obtain the final product.

[0015] More preferably, the preparation method of the composite modified peptide includes the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal are mixed with water at a mass ratio of 6-7:3-4, with a mass concentration of 15-20%. The mixture is sterilized at 110-120℃ for 15-20 minutes, and inoculated with Lactobacillus plantarum and Bacillus subtilis, with a live bacteria ratio of (2-3):1. The total inoculation amount is 2%-5% of the fermentation liquid volume. The mixture is fermented at 35-40℃ for 24-36 hours, and then heated to 85-90℃ for 8-10 minutes to inactivate the bacteria, thus obtaining the fermentation liquid. S2: Cool the fermentation broth to 45-55℃, adjust the pH to 8-8.5, add alkaline protease and hydrolyze for 1.5-2.5 hours. The amount of alkaline protease added is 1800-2000 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Then adjust the pH to 7.0-7.5, add flavor protease and specific leucine aminopeptidase. The amount of flavor protease added is 400-600 U / g, and the amount of specific leucine aminopeptidase added is 900-1100 U / g, both calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Continue hydrolysis for 2-3 hours, then heat to 85-90℃ to inactivate the enzymes for 8-10 minutes to obtain the hydrolysate. S3: Concentrate the enzymatic hydrolysate to a solid content of 25-30%, then add 1.5-2.0 times the dry matter weight of the enzymatic hydrolysate of galactooligosaccharides (DP3-5), adjust the pH to 7.5-8.0, react at 60-65℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction, dry, and the product is obtained.

[0016] Preferably, the vitamins include fat-soluble vitamins and water-soluble vitamins.

[0017] The fat-soluble vitamins include at least one of vitamin A, vitamin D, and vitamin E; the water-soluble vitamins include at least one of vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, sodium L-ascorbate, and biotin.

[0018] Preferably, the mineral is selected from at least one of magnesium oxide, ferric pyrophosphate, zinc sulfate, and sodium selenite.

[0019] Preferably, the excipients are glycine, citric acid, microcrystalline cellulose, and low-acyl gellan gum.

[0020] Preferably, the ingredients, by weight, include the following raw materials: 8-15 parts of compound protein, 2-6 parts of compound modified peptide, 2-5 parts of medium- and long-chain triglycerides, 2-5 parts of sunflower seed oil, 0.5-2 parts of fish oil, 0.5-1.5 parts of phytosterol esters, 0.05-0.2 parts of vitamins, 0.1-0.5 parts of minerals, 59-85 parts of water, and 0.05-0.1 parts of excipients.

[0021] More preferably, the ingredients, by weight, include the following: 8-15 parts of complex protein, 2-6 parts of complex modified peptide, 2-5 parts of medium- and long-chain triglycerides, 2-5 parts of sunflower seed oil, 0.5-2 parts of fish oil, 0.5-1.5 parts of phytosterol esters, 0.05-0.2 parts of vitamins, 0.1-0.5 parts of minerals, 59.94-84.92 parts of water, and 0.06-0.08 parts of excipients.

[0022] Preferably, the excipients include glycine, citric acid, microcrystalline cellulose, and low-acyl gellan gum.

[0023] Preferably, the mass ratio of glycine, citric acid, microcrystalline cellulose and low-acyl gellan gum is 0.01:(0.01-0.03):(0.08-0.12):(0.01-0.03).

[0024] Secondly, the present invention provides a method for preparing the above-mentioned high-protein nutrient solution composition, comprising the following steps: Step 1: Mix minerals, glycine, and citric acid with water to carry out a chelation reaction to obtain a mineral chelate solution; mix fat-soluble vitamins with medium- and long-chain triglycerides, sunflower seed oil, fish oil, and phytosterol esters, and heat to dissolve to obtain a composite oil phase; mix water-soluble vitamins with water to dissolve to obtain a water-soluble vitamin solution. Step 2: Mix the composite protein, composite modified peptide, microcrystalline cellulose with the mineral chelate solution and water-soluble vitamin solution from Step 1, add water, and shear disperse to obtain an aqueous phase; add the composite oil phase to the aqueous phase and perform high-shear emulsification to obtain the colostrum; Step 3: Take 15%-25% of the total volume of the colostrum obtained in Step 2 and subject it to high-pressure microfluidic treatment to obtain nano-emulsion; mix the nano-emulsion with the remaining colostrum and perform two-stage high-pressure homogenization to obtain a homogenized emulsion. Step 4: Add low-acyl gellan gum to the homogenized emulsion, shear hydration, and then perform temperature control cooling treatment. Subsequently, perform ultra-high temperature instantaneous sterilization, cool and fill to obtain the high-protein nutrient solution composition.

[0025] Preferably, in step 1, the chelation reaction is carried out at a temperature of 45-55°C for 30-40 minutes; the heating and dissolving temperature is 55-65°C.

[0026] Preferably, in step 2, the rotation speed of the shear dispersion is 8000-10000 r / min, and the time is 10-15 min; the rotation speed of the high-shear emulsification is 8000-12000 r / min, the time is 5-10 min, and the emulsification temperature is 55-65℃.

[0027] Preferably, in step 3, the pressure of the high-pressure microjet treatment is 100-120 MPa, and the treatment is repeated 2-4 times; the temperature of the two-stage high-pressure homogenization is 55-65℃, the pressure of the first-stage homogenization is 25-35 MPa, and the pressure of the second-stage homogenization is 5-10 MPa.

[0028] Preferably, in step 4, the temperature of the shear hydration is 75-85℃, the rotation speed is 3000-5000 r / min, and the time is 15-20 min; the temperature control and cooling process is: slowly cooling to 20-30℃ at a cooling rate of 0.5-1.5℃ / min, while maintaining continuous stirring at 200-400 r / min during the cooling process; the temperature of the ultra-high temperature instantaneous sterilization is 137-139℃, and the time is 4-6 s; the cooling and filling process is aseptic cold filling after cooling to below 25℃.

[0029] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention utilizes a specific preparation method to obtain a composite protein, which, through specific enzymatic hydrolysis and ferulic acid-mediated laccase-directed cross-linking, reconstructs asymmetric heterologous protein aggregates of "flexible plant protein chains - rigid whey protein spheres" at the molecular level. Combined with a staged emulsification process of "high-pressure microfluidic jet + two-stage high-pressure homogenization" and specific excipients, a specific "micron-nano" bimodal particle size distribution emulsion is constructed. This solves the technical problems of easy demulsification and poor stability.

[0030] 2. This invention uses deep-sea fish skin collagen and defatted walnut meal as substrates. First, it employs co-fermentation with *Lactobacillus plantarum* and *Bacillus subtilis* for natural flavor enhancement and pre-degradation. Subsequently, it utilizes a multi-enzyme cascade enzymatic hydrolysis, and leverages a specific leucine aminopeptidase to precisely remove the hydrophobic bitter amino acids at the peptide terminals, eradicating bitterness at the molecular level. Furthermore, galactooligosaccharides are covalently grafted onto the peptide molecules, significantly improving the product's sensory quality and thermal stability.

[0031] 3. This invention addresses the issues of mineral precipitation and catalytic oxidation by chelating minerals with glycine and citric acid, forming a chelate with an extremely stable spatial structure, completely isolating the catalytic oxidation of lipids by free metal ions. Simultaneously, fat-soluble vitamins are dissolved in the composite oil phase, further improving product stability and effectively preventing nutritional degradation and off-flavors. Detailed Implementation

[0032] Terminology and Declarations of this Invention: 1. Articles “a,” “a kind,” and “the”: These include plural objects unless otherwise explicitly specified as a single (kind) object.

[0033] 2. Numerical Range: Unless otherwise expressly stated, all ranges or ratios disclosed herein shall be construed as including any and all subranges or subratios contained herein. For example, a stated range or ratio of 1 to 30 shall be considered to be included between the minimum value of 1 and the maximum value of 30, and includes any subranges or subratios, integers, decimals, or subranges or subratios consisting of integers or decimals, including endpoints.

[0034] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0035] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention were obtained through conventional commercial means. Unless otherwise specified, all contents mentioned below are mass contents. Unless otherwise specified, it is understood that the process was carried out at room temperature.

[0036] In the following examples, some of the reagents were sourced from the following table: Table 1

[0037] The preparation method of the complex protein consists of the following steps: (1) Pea protein isolate and chickpea protein isolate are mixed with water, with the mass percentage of pea protein isolate and chickpea protein isolate in water being 10%. The pH is adjusted to 7.8, and alkaline protease is added and enzymatically hydrolyzed at 50°C for 30 min. The amount of alkaline protease added is 500 U / g, and the amount added is based on the total dry matter mass of pea protein isolate and chickpea protein isolate. The temperature is raised to 90°C to inactivate the enzyme for 8 min, and then cooled to 40°C to obtain protein solution. (2) The protein solution from step (1) is mixed with whey protein isolate, and ferulic acid and laccase at a mass of 0.8% of the total dry matter of the protein in the system and 6 U / g are added. The amount of laccase added is based on the mass of the total dry matter of the protein in the system. The pH is adjusted to 6.5, and the mixture is stirred at 35°C for 3 hours. The temperature is then raised to 90°C to inactivate the enzyme for 8 minutes. The mixture is then dried to obtain the composite protein.

[0038] The mass ratio of whey protein isolate in step (2), pea protein isolate in step (1), and chickpea protein isolate in step (1) is 4:3:3.

[0039] The prepared composite protein was used in the examples and some comparative examples of the present invention.

[0040] The method for preparing the composite modified peptide involves the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal were mixed with water at a mass ratio of 6.5:3.5, with a mass concentration of 18%. The mixture was sterilized at 115°C for 18 minutes, and then inoculated with Lactobacillus plantarum and Bacillus subtilis, with a live bacteria ratio of 2.5:1. The total inoculation amount was 4% of the fermentation liquid volume. The mixture was fermented at 35°C for 30 hours, and then heated to 90°C for 8 minutes to inactivate the bacteria, thus obtaining the fermentation liquid. S2: Cool the fermentation broth to 50℃, adjust the pH to 8.2, add alkaline protease for 2 hours of enzymatic hydrolysis. The amount of alkaline protease added is 2000 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Then adjust the pH to 7.2, add flavor protease and specific leucine aminopeptidase. The amount of flavor protease added is 500 U / g, and the amount of specific leucine aminopeptidase added is 1000 U / g, both calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Continue enzymatic hydrolysis for 2.5 hours, then heat to 90℃ for 8 minutes to inactivate the enzymes, obtaining the enzymatic hydrolysate. S3: After concentrating the enzymatic hydrolysate to a solid content of 28%, add 1.8 times the dry matter weight of the enzymatic hydrolysate, add galactooligosaccharides (DP3-5), adjust the pH to 8.0, react at 60℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction, dry, and the product is obtained.

[0041] The prepared composite modified peptides were used in the examples and some comparative examples of this invention.

[0042] Example 1 A high-protein nutrient solution composition comprises the following raw materials: 12 parts of complex protein, 4 parts of complex modified peptides, 4 parts of medium- and long-chain triglycerides, 3 parts of sunflower seed oil, 1.5 parts of fish oil, 1 part of phytosterol esters, 0.15 parts of vitamins (vitamin A, vitamin B1, vitamin B12, niacin, biotin, and vitamin E in a mass ratio of 2:1:1:3:1:1), 0.3 parts of minerals (zinc sulfate and ferric pyrophosphate in a mass ratio of 1:1), 0.005 parts of glycine, 0.01 parts of citric acid, 0.05 parts of microcrystalline cellulose, 0.01 parts of low-acyl gellan gum, and 75.92 parts of water.

[0043] The preparation method consists of the following steps: Step 1: Mix minerals, glycine, and citric acid with water (the mass of water is 1 / 2 of the mass of water in the formula) to carry out a chelation reaction to obtain a mineral chelate solution; mix fat-soluble vitamins with medium- and long-chain triglycerides, sunflower seed oil, fish oil, and phytosterol esters, and heat to dissolve to obtain a composite oil phase; mix water-soluble vitamins with water (the mass of water is 1 / 5 of the mass of water in the formula) to dissolve to obtain a water-soluble vitamin solution; the chelation reaction temperature is 50℃ and the time is 35 min; the heating and dissolving temperature is 60℃; Step 2: Mix the composite protein, composite modified peptide, microcrystalline cellulose with the mineral chelate solution and water-soluble vitamin solution from Step 1, add the remaining water, and shear disperse to obtain an aqueous phase; add the composite oil phase to the aqueous phase and perform high-shear emulsification to obtain a primary emulsion; the shear dispersion speed is 9000 r / min and the time is 12 min; the high-shear emulsification speed is 10000 r / min and the time is 8 min, and the emulsification temperature is 60℃.

[0044] Step 3: Take 20% of the total volume of the colostrum obtained in Step 2 and perform high-pressure microjets to obtain nano-emulsions; mix the nano-emulsions with the remaining colostrum and perform two-stage high-pressure homogenization to obtain a homogenized emulsion; the pressure of the high-pressure microjets is 110 MPa, and the process is repeated 3 times; the temperature of the two-stage high-pressure homogenization is 60℃, the pressure of the first-stage homogenization is 30 MPa, and the pressure of the second-stage homogenization is 8 MPa.

[0045] Step 4: Add low-acyl gellan gum to the homogenized emulsion, shear and hydrate at 80°C and 4000 r / min for 18 min, then perform temperature-controlled cooling treatment, slowly cooling to 25°C at a cooling rate of 1°C / min, while maintaining continuous stirring at 300 r / min during the cooling process. Subsequently, perform ultra-high temperature instantaneous sterilization at a temperature of 138°C for 5 s; cool to below 25°C and perform aseptic cold filling to obtain the high-protein nutrient solution composition.

[0046] Example 2 A high-protein nutrient solution composition comprises the following raw materials: 8 parts of complex protein, 6 parts of complex modified peptides, 2 parts of medium- and long-chain triglycerides, 5 parts of sunflower seed oil, 0.5 parts of fish oil, 1.5 parts of phytosterol esters, 0.05 parts of vitamins (vitamin D, vitamin B2, vitamin B6, folic acid, sodium L-ascorbate, and vitamin E in a mass ratio of 2:1:1:1:1:1), 0.5 parts of minerals (magnesium oxide), 0.004 parts of glycine, 0.012 parts of citric acid, 0.032 parts of microcrystalline cellulose, 0.012 parts of low-acyl gellan gum, and 59.94 parts of water.

[0047] The preparation method consists of the following steps: Step 1: Mix minerals, glycine, and citric acid with water (the mass of water is 1 / 2 of the mass of water in the formula) to carry out a chelation reaction to obtain a mineral chelate solution; mix fat-soluble vitamins with medium- and long-chain triglycerides, sunflower seed oil, fish oil, and phytosterol esters, and heat to dissolve to obtain a composite oil phase; mix water-soluble vitamins with water (the mass of water is 1 / 5 of the mass of water in the formula) to dissolve to obtain a water-soluble vitamin solution; the chelation reaction temperature is 45℃ and the time is 40 min; the heating and dissolving temperature is 65℃; Step 2: Mix the composite protein, composite modified peptide, microcrystalline cellulose with the mineral chelate solution and water-soluble vitamin solution from Step 1, add the remaining water, and shear disperse to obtain an aqueous phase; add the composite oil phase to the aqueous phase and perform high-shear emulsification to obtain a primary emulsion; the shear dispersion speed is 8000 r / min and the time is 15 min; the high-shear emulsification speed is 12000 r / min and the time is 5 min, and the emulsification temperature is 55℃.

[0048] Step 3: Take 15% of the total volume of the colostrum obtained in Step 2 and perform high-pressure microjets to obtain nano-emulsions; mix the nano-emulsions with the remaining colostrum and perform two-stage high-pressure homogenization to obtain a homogenized emulsion; the pressure of the high-pressure microjets is 120 MPa, and the process is repeated twice; the temperature of the two-stage high-pressure homogenization is 55℃, the pressure of the first-stage homogenization is 35 MPa, and the pressure of the second-stage homogenization is 5 MPa.

[0049] Step 4: Add low-acyl gellan gum to the homogenized emulsion, shear hydrate at 75°C and 5000 r / min for 15 min, then perform temperature-controlled cooling treatment, slowly cooling to 20°C at a cooling rate of 1.5°C / min, while maintaining continuous stirring at 400 r / min during the cooling process. Subsequently, perform ultra-high temperature instantaneous sterilization at a temperature of 137°C for 6 seconds; cool to below 25°C and perform aseptic cold filling to obtain the high-protein nutrient solution composition.

[0050] Example 3 A high-protein nutrient solution composition comprises the following raw materials: 15 parts of complex protein, 2 parts of complex modified peptide, 5 parts of medium- and long-chain triglycerides, 2 parts of sunflower seed oil, 2 parts of fish oil, 0.5 parts of phytosterol esters, 0.2 parts of vitamins (vitamin D, vitamin B2, vitamin B6, pantothenic acid, sodium L-ascorbate, and vitamin E in a mass ratio of 2:1:1:1:1:1), 0.1 parts of minerals (sodium selenite), 0.005 parts of glycine, 0.005 parts of citric acid, 0.067 parts of microcrystalline cellulose, 0.005 parts of low-acyl gellan gum, and 84.92 parts of water.

[0051] The preparation method consists of the following steps: Step 1: Mix minerals, glycine, and citric acid with water (the mass of water is 1 / 2 of the mass of water in the formula) to carry out a chelation reaction to obtain a mineral chelate solution; mix fat-soluble vitamins with medium- and long-chain triglycerides, sunflower seed oil, fish oil, and phytosterol esters, and heat to dissolve to obtain a composite oil phase; mix water-soluble vitamins with water (the mass of water is 1 / 5 of the mass of water in the formula) to dissolve to obtain a water-soluble vitamin solution; the chelation reaction temperature is 55℃ and the time is 3 minutes; the heating and dissolving temperature is 55℃. Step 2: Mix the composite protein, composite modified peptide, microcrystalline cellulose with the mineral chelate solution and water-soluble vitamin solution from Step 1, add the remaining water, and shear disperse to obtain an aqueous phase; add the composite oil phase to the aqueous phase and perform high-shear emulsification to obtain a primary emulsion; the shear dispersion speed is 10000 r / min and the time is 10 min; the high-shear emulsification speed is 8000 r / min and the time is 10 min, and the emulsification temperature is 65℃.

[0052] Step 3: Take 25% of the total volume of the colostrum obtained in Step 2 and perform high-pressure microjets to obtain nano-emulsions; mix the nano-emulsions with the remaining colostrum and perform two-stage high-pressure homogenization to obtain a homogenized emulsion; the pressure of the high-pressure microjets is 100 MPa, and the process is repeated 4 times; the temperature of the two-stage high-pressure homogenization is 65℃, the pressure of the first-stage homogenization is 25 MPa, and the pressure of the second-stage homogenization is 10 MPa.

[0053] Step 4: Add low-acyl gellan gum to the homogenized emulsion, shear and hydrate at 85°C and 3000 r / min for 20 min, then perform temperature-controlled cooling treatment, slowly cooling to 30°C at a cooling rate of 0.5°C / min, while maintaining continuous stirring at 200 r / min during the cooling process. Subsequently, perform ultra-high temperature instantaneous sterilization at a temperature of 139°C for 4 seconds; cool to below 25°C and perform aseptic cold filling to obtain the high-protein nutrient solution composition.

[0054] Comparative Example 1 High-protein milk prepared according to Example 1 of Chinese patent application CN115644429A.

[0055] Comparative Example 2 A high-protein nutrient solution composition comprises the following raw materials: 15.5 parts of complex protein, 1 part of complex modified peptide, 1 part of medium- and long-chain triglycerides, 1 part of sunflower seed oil, 5 parts of fish oil, 2 parts of phytosterol esters, 0.3 parts of vitamins (vitamin A, vitamin B1, vitamin B12, niacin, biotin, and vitamin E in a mass ratio of 2:1:1:3:1:1), 0.05 parts of minerals (zinc sulfate and ferric pyrophosphate in a mass ratio of 1:1), 0.005 parts of glycine, 0.01 parts of citric acid, 0.05 parts of microcrystalline cellulose, 0.01 parts of low-acyl gellan gum, and 75.92 parts of water.

[0056] The preparation method is the same as in Example 1.

[0057] Comparative Example 3 A high-protein nutrient solution composition, compared with Example 1, except that the complex protein is replaced with a mixture of whey protein isolate, pea protein isolate and chickpea protein isolate in a mass ratio of 4:3:3.

[0058] Comparative Example 4 A high-protein nutrient solution composition, compared with Example 1, except that the composite modified peptide is replaced with the Peili peptide of Chinese patent application CN109744317A.

[0059] Comparative Example 5 A high-protein nutrient solution composition, compared with Example 1, differs only in the method of preparing the complex protein: The preparation method of the complex protein consists of the following steps: Whey protein, pea protein isolate, and chickpea protein isolate in a mass ratio of 4:3:3 were mixed with water, with the pea protein isolate and chickpea protein isolate comprising 10% by mass in the water. The pH was adjusted to 7.8, and alkaline protease was added and enzymatically hydrolyzed at 50°C for 30 minutes. The amount of alkaline protease added was 500 U / g, based on the total dry matter mass of the pea protein isolate and chickpea protein isolate. The enzyme was inactivated by heating to 90°C for 8 minutes, and then cooled to 40°C to obtain a protein solution. The solution was then dried to obtain the complex protein.

[0060] The rest is the same as in Example 1.

[0061] Comparative Example 6 A high-protein nutrient solution composition, compared with Example 1, differs only in the preparation method of the composite modified peptide: The method for preparing the composite modified peptide involves the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal were mixed with water at a mass ratio of 6.5:3.5, with a mass concentration of 18%. The mixture was sterilized at 115℃ for 18 minutes, inoculated with Lactobacillus plantarum, with a total inoculation amount of 4% of the fermentation liquid volume, and fermented at 35℃ for 30 hours. The mixture was then heated to 90℃ for 8 minutes to inactivate the bacteria, thus obtaining the fermentation liquid. S2: Cool the fermentation broth to 50℃, adjust the pH to 8.2, add alkaline protease for 2 hours of enzymatic hydrolysis. The amount of alkaline protease added is 2000 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Then adjust the pH to 7.2, add flavor protease at a rate of 1500 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Continue enzymatic hydrolysis for 2.5 hours, then heat to 90℃ to inactivate the enzyme for 8 minutes to obtain the enzymatic hydrolysate. S3: After concentrating the enzymatic hydrolysate to a solid content of 28%, add 1.8 times the dry matter weight of the enzymatic hydrolysate, add galactooligosaccharides (DP3-5), adjust the pH to 8.0, react at 60℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction, dry, and the product is obtained.

[0062] The rest is the same as in Example 1.

[0063] Comparative Example 7 A high-protein nutrient solution composition, compared with Example 1, differs only in the preparation method of the composite modified peptide: The method for preparing the composite modified peptide involves the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal were mixed with water at a mass ratio of 6.5:3.5, with a mass concentration of 18%. The mixture was sterilized at 115℃ for 18 minutes, inoculated with Bacillus subtilis, and the total inoculation amount was 4% of the fermentation liquid volume. The mixture was fermented at 35℃ for 30 hours, and then heated to 90℃ for 8 minutes to inactivate the bacteria, thus obtaining the fermentation liquid. S2: Cool the fermentation broth to 50℃, adjust the pH to 8.2, add alkaline protease for 2 hours of enzymatic hydrolysis. The amount of alkaline protease added is 2000 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Then adjust the pH to 7.2, add specific leucine aminopeptidase at a concentration of 1500 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Continue enzymatic hydrolysis for 2.5 hours, then heat to 90℃ for 8 minutes to inactivate the enzyme, obtaining the enzymatic hydrolysate. S3: After concentrating the enzymatic hydrolysate to a solid content of 28%, add 1.8 times the dry matter weight of the enzymatic hydrolysate, add galactooligosaccharides (DP3-5), adjust the pH to 8.0, react at 60℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction, dry, and the product is obtained.

[0064] The rest is the same as in Example 1.

[0065] Comparative Example 8 A high-protein nutrient solution composition, compared with Example 1, differs only in the preparation method of the composite modified peptide: The method for preparing the composite modified peptide involves the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal were mixed with water at a mass ratio of 6.5:3.5, with a mass concentration of 18%. The mixture was sterilized at 115°C for 18 minutes, and then inoculated with Lactobacillus plantarum and Bacillus subtilis, with a live bacteria ratio of 2.5:1. The total inoculation amount was 4% of the fermentation liquid volume. The mixture was fermented at 35°C for 30 hours, and then heated to 90°C for 8 minutes to inactivate the bacteria, thus obtaining the fermentation liquid. S2: Cool the fermentation broth to 50℃, adjust the pH to 8.2, add alkaline protease for 2 hours of enzymatic hydrolysis. The amount of alkaline protease added is 2000 U / g, calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Then adjust the pH to 7.2, add flavor protease and specific leucine aminopeptidase. The amount of flavor protease added is 500 U / g, and the amount of specific leucine aminopeptidase added is 1000 U / g, both calculated based on the total dry matter mass of deep-sea fish skin collagen and defatted walnut meal. Continue enzymatic hydrolysis for 2.5 hours, then heat to 90℃ for 8 minutes to inactivate the enzymes, obtaining the enzymatic hydrolysate. Dry to obtain the final product.

[0066] The rest is the same as in Example 1.

[0067] Test Example 1 Stability test: 1. Testing Method: (1) Centrifugation sedimentation rate: Accurately weigh the sample (about 50g) into a centrifuge tube (recorded as the total mass of the sample and the centrifuge tube). m 1) Centrifuge at 4000 r / min and 25℃ for 15 min. After centrifugation, carefully discard the supernatant and weigh the precipitate and the total mass of the centrifuge tube (recorded as ). m 2), and weigh the empty centrifuge tubes in advance (recorded as ). m 0).

[0068] Centrifugal sedimentation rate (%) = ( m 2 m 0) / ( m 1 m 0)×100%.

[0069] (2) Particle size: The average particle size (Z-average) of the sample was determined using a Malvern laser particle size analyzer (ZetasizerNanoZS). The sample was appropriately diluted with deionized water before the test, and the test temperature was 25℃.

[0070] (3) Fat floating: After centrifuging, take out the centrifuge tube, place it vertically upright, observe and measure the height (mm) of the upper fat layer.

[0071] (4) Viscosity: The apparent viscosity of the sample was measured using a Brookfield rotational viscometer at 25°C and a rotor speed of 60 r / min.

[0072] 2. Evaluation Criteria Centrifugal sedimentation rate: The reasonable range is 0.1% - 2.0%. The smaller the value, the better the suspension stability of the system.

[0073] Particle size: The reasonable range is 200-600 nm. The smaller the particle size and the narrower the distribution, the better the emulsification effect and the more stable the system.

[0074] Fat floating: A small amount / slight floating (height ≤ 2mm) is acceptable; a large amount of floating (height > 2mm) is unacceptable.

[0075] Viscosity: The reasonable range is 20 - 40 mPa·s. Within this range, the taste is smooth and phase separation is not easy to occur.

[0076] 3. The test results are shown in Table 2 below.

[0077] Table 2. Stability test results of each high-protein nutrient solution composition.

[0078] As shown in Table 2, the centrifugal sedimentation rate, particle size, and viscosity of Examples 1-3 are all within the optimal range, and no fat floats to the surface after standing for 30 days, demonstrating excellent physical stability. This is due to the excellent emulsifying and water-holding properties of the composite protein and composite modified peptide in this invention, as well as the nanoscale fine emulsion structure formed by the specific process.

[0079] Test Example 2 Sensory evaluation test 1. Testing Method: Twenty professionally trained sensory evaluators (half male and half female, aged 20-40) were recruited to conduct blind testing in an evaluation room at room temperature (25℃), with sufficient light and no odor. Each evaluator tasted approximately 30mL of sample, rinsing their mouth with purified water between tastings.

[0080] 2. The evaluation criteria adopt a 100-point system, and scores are given from four dimensions: color, smell, taste and texture. The specific criteria are shown in Table 3.

[0081] Table 3 Sensory Evaluation Criteria

[0082] 3. The test results are shown in Table 4 below.

[0083] Table 4. Sensory evaluation scores (average scores) of each high-protein nutrient solution composition.

[0084] As shown in Table 4, Examples 1-3 all scored above 93 points, demonstrating excellent performance. This invention cross-links the complex protein with alkaline protease and laccase, and prepares a complex modified peptide using multi-enzyme combined enzymatic hydrolysis and Maillard reaction. This effectively removes the beany taste of peas and chickpeas, as well as the bitter peptides produced by enzymatic hydrolysis. Simultaneously, the Maillard reaction imparts an appealing color and nutty aroma to the product.

[0085] Test Example 3 Anti-fatigue animal experiments 1. Testing Method (1) Experimental animals and grouping: 130 healthy male SPF-grade Kunming mice weighing 18-22g were selected. After 3 days of acclimatization feeding, they were randomly divided into 13 groups (blank control group, Example 1-3 groups, and Comparative Example 1-8 groups), with 10 mice in each group.

[0086] (2) Administration method: Each treatment group was given the corresponding high-protein nutrient solution by gavage at a dose of 10 mL / kg·bw, while the blank control group was given an equal volume of physiological saline by gavage. The gavage was performed once a day for 30 consecutive days.

[0087] (3) Weighted swimming experiment: 30 minutes after the last gavage, a lead wire weighing 5% of the mouse's body weight was placed on the base of the mouse's tail, and the mouse was placed in a swimming tank with a water depth of 30 cm and a water temperature of 25±1℃. The time from when the mouse entered the water until it sank to the bottom of the water and could not float to the surface for 10 seconds was recorded as the weighted swimming time.

[0088] (4) Biochemical index determination: After the mice finished swimming, the eyeballs were immediately removed and blood was collected. The serum was separated by centrifugation at 3000 r / min for 10 min. The serum urea nitrogen (BUN) and blood lactate (BLA) content were determined by a fully automated biochemical analyzer. The mice were then euthanized by cervical dislocation, and the liver was quickly removed. The liver glycogen content was determined by anthrone colorimetric method.

[0089] 2. Evaluation Criteria Swimming time with added weight: The longer the time, the stronger the ability to resist fatigue.

[0090] Serum blood urea nitrogen (BUN): a protein metabolite. The lower the value, the less protein is broken down for energy, and the faster the fatigue recovery.

[0091] Blood lactate (BLA): a product of anaerobic glycolysis. The lower the value, the less lactate accumulates, and the better the anti-fatigue effect.

[0092] Glycogen: an energy reserve substance. The higher the value, the more abundant the energy reserves and the stronger the ability to resist fatigue.

[0093] 3. Test Results Table 5. Results of weight-bearing swimming time and biochemical indicators for each group of mice ( ±s, n=10)

[0094] Note: Compared with the blank control group % P<0.05, %% P<0.01.

[0095] Compared with Example 1 group,# P<0.05, ## P<0.01.

[0096] As shown in Table 5, compared with the blank control group, the weight-bearing swimming time of mice in Examples 1-3 was significantly prolonged (P<0.01), and the serum BUN and BLA levels were significantly decreased (P<0.01), while the liver glycogen reserves were significantly increased (P<0.01). This indicates that the high-protein nutrient solution prepared in this invention can significantly improve the body's exercise endurance, effectively delay the onset of fatigue, and accelerate fatigue elimination. Its core mechanism lies in the fact that the specially formulated "composite modified peptide" of this invention releases a large amount of highly active small molecule peptides rich in branched-chain amino acids (BCAAs) through multi-enzyme directed enzymatic decomposition, which are easily and rapidly absorbed by the intestines and directly participate in muscle energy metabolism; at the same time, the Maillard reaction products endow the system with strong antioxidant activity, which can efficiently scavenge free radicals generated during exercise and reduce muscle cell damage.

[0097] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A high-protein nutrient solution composition, characterized in that, The product comprises the following raw materials: compound protein, compound modified peptide, medium- and long-chain triglycerides, sunflower seed oil, fish oil, phytosterol esters, vitamins, minerals, water, and excipients; the compound protein is prepared by enzymatic hydrolysis of pea protein isolate, chickpea protein isolate, and whey protein isolate; the compound modified peptide is prepared by fermentation, enzymatic hydrolysis, and glycosylation grafting modification of deep-sea fish skin collagen and defatted walnut meal; the excipients are selected from at least one of glycine, citric acid, microcrystalline cellulose, and low-acyl gellan gum.

2. The high-protein nutrient solution composition according to claim 1, characterized in that, The method for preparing the complex protein includes the following steps: (1) Mix pea protein isolate and chickpea protein isolate with water, add alkaline protease to hydrolyze for 30-40 minutes, inactivate the enzyme, and obtain protein solution; (2) Mix the protein solution with whey protein isolate, add ferulic acid and laccase for enzymatic hydrolysis and enzyme inactivation, and the protein is obtained.

3. The high-protein nutrient solution composition according to claim 2, characterized in that, The mass ratio of whey protein isolate, pea protein isolate, and chickpea protein isolate is 3-5:2-4:2-4.

4. The high-protein nutrient solution composition according to claim 1, characterized in that, The preparation method of the composite modified peptide includes the following steps: S1: Deep-sea fish skin collagen and defatted walnut meal are mixed with water, sterilized, inoculated with Lactobacillus plantarum and Bacillus subtilis, fermented, inactivated, and fermented to obtain fermentation broth; S2: Cool the fermentation broth to 45-55℃, adjust the pH to 8-8.5, add alkaline protease and hydrolyze for 1.5-2.5h; then adjust the pH to 7.0-7.5, add flavor protease and specific leucine aminopeptidase, continue hydrolysis for 2-3h, inactivate the enzyme, and obtain the hydrolysate. S3: After concentrating the enzymatic hydrolysate, add 1.5-2.0 times the dry matter weight of the enzymatic hydrolysate, add galactooligosaccharides (DP3-5), adjust the pH to 7.5-8.0, react at 60-65℃, and monitor the absorbance of the reaction solution at 294nm in real time. When A294 reaches 0.40-0.55, stop the reaction to obtain the final product.

5. The high-protein nutrient solution composition according to claim 1, characterized in that, The vitamins include fat-soluble vitamins and water-soluble vitamins.

6. The high-protein nutrient solution composition according to claim 5, characterized in that, The fat-soluble vitamins include at least one of vitamin A, vitamin D, and vitamin E; the water-soluble vitamins include at least one of vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, sodium L-ascorbate, and biotin.

7. The high-protein nutrient solution composition according to claim 1, characterized in that, The mineral is selected from at least one of magnesium oxide, ferric pyrophosphate, zinc sulfate, and sodium selenite.

8. The high-protein nutrient solution composition according to claim 1, characterized in that, By weight, it includes the following raw materials: 8-15 parts of compound protein, 2-6 parts of compound modified peptide, 2-5 parts of medium and long chain triglycerides, 2-5 parts of sunflower seed oil, 0.5-2 parts of fish oil, 0.5-1.5 parts of phytosterol esters, 0.05-0.2 parts of vitamins, 0.1-0.5 parts of minerals, 59-85 parts of water, and 0.05-0.1 parts of excipients.

9. The high-protein nutrient solution composition according to claim 1, characterized in that, The excipients are glycine, citric acid, microcrystalline cellulose, and low-acyl gellan gum in a mass ratio of 0.01:(0.01-0.03):(0.08-0.12):(0.01-0.03).

10. A method for preparing the high-protein nutrient solution composition according to any one of claims 1-9, characterized in that, Includes the following steps: Step 1: Mix minerals, glycine, and citric acid with water to carry out a chelation reaction to obtain a mineral chelate solution; mix fat-soluble vitamins with medium- and long-chain triglycerides, sunflower seed oil, fish oil, and phytosterol esters, and heat to dissolve to obtain a composite oil phase; mix water-soluble vitamins with water to dissolve to obtain a water-soluble vitamin solution. Step 2: Mix the composite protein, composite modified peptide, microcrystalline cellulose with the mineral chelate solution and water-soluble vitamin solution from Step 1, add water, and shear disperse to obtain an aqueous phase; add the composite oil phase to the aqueous phase and perform high-shear emulsification to obtain the colostrum; Step 3: Take 15%-25% of the total volume of the colostrum obtained in Step 2 and subject it to high-pressure microfluidic treatment to obtain nano-emulsion; mix the nano-emulsion with the remaining colostrum and perform two-stage high-pressure homogenization to obtain a homogenized emulsion. Step 4: Add low-acyl gellan gum to the homogenized emulsion, shear hydration, and then perform temperature control cooling treatment. Subsequently, perform ultra-high temperature instantaneous sterilization, cool and fill to obtain the high-protein nutrient solution composition.

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