High-absorption type egg white peptide sports protein powder rich in branched chain amino acid and preparation method of high-absorption type egg white peptide sports protein powder

Abstract

The invention discloses high-absorption type egg white peptide sports protein powder rich in branched chain amino acid and a preparation method thereof, and relates to the field of foods for special diets. The preparation method of the sports protein powder comprises the following step of mixing the egg white peptide containing the DKVVDVD sequence, the whey protein powder, the skim milk powder, the strawberry essence, the sucralose, the composite mineral substances and the composite vitamins step by step according to a specific sequence. Compared with commercially available products, the product disclosed by the invention has no significant difference in sensory evaluation, but the content of branched chain amino acid is increased by 64.42%, and the particle size and the PDI value are respectively reduced by 28.20% and 17.62%, so that the product has higher absorption efficiency. The product is simple and convenient in preparation process, is nutritional and delicious, can meet the nutritional requirements of sports crowds, remarkably improves the nutrient utilization degree, and achieves the functional goals of promoting muscle growth and relieving sports fatigue.

Description

Technical Field

[0001] The invention belongs to the field of special dietary foods, and particularly relates to a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids and a preparation method thereof. Background Art

[0002] In recent years, with the rise of a nationwide fitness craze, the market for sports nutrition foods has continued to expand. Protein, as a core nutrient in sports nutrition foods, promotes muscle repair and replenishes energy, playing a key role in nutritional supplementation for athletes. However, despite the diverse range of sports nutrition products, they generally face the problem of raw material homogeneity, especially the underdevelopment of domestic protein resources beyond traditional protein sources such as whey protein and soy protein. For example, as the world's largest producer of eggs, egg whites, with their high protein content and low fat content, are a promising source of high-quality protein. However, their development and application in sports nutrition foods has not yet become widespread. Furthermore, the particle size and polymer dispersibility index (PDI) of sports protein powder solutions are relatively high, resulting in uneven dispersion of protein particles in the solution and poor stability. These characteristics significantly slow the dissolution rate of protein in the gastrointestinal tract, hindering its effective contact with intestinal absorption sites, and significantly reducing the body's absorption efficiency of protein powder. Therefore, it is of great significance to develop high-quality protein resources and create highly absorbable protein powders that meet the nutritional needs of athletes.

[0003] Branched-chain amino acids (leucine, isoleucine and valine) are essential amino acids for the human body. They stimulate muscle protein synthesis and improve muscle mass during the body's metabolic process. However, the human body lacks the key enzymes to synthesize these amino acids and must be obtained exogenously through diet or nutritional supplements. It is worth noting that free branched-chain amino acids are easily directly decomposed by pepsin during digestion, resulting in a significant decrease in their bioavailability. The introduction of bioactive peptides provides a solution to the above problems. Bioactive peptides are low-molecular-weight fragments produced by processes such as enzymatic hydrolysis of proteins. They can not only prolong the action time of amino acids and improve their absorption efficiency, but also have multiple physiological functions such as anti-inflammatory and antioxidant. Therefore, the preparation of bioactive peptides rich in branched-chain amino acids and their application in the field of sports nutrition has broad prospects.

[0004] This patent has fundamentally different protection directions from the already authorized "A composite fitness powder for rapid recovery of physical strength and muscle growth" (Patent No.: CN201710701559.0) and "A sports nutritional supplement and its preparation method" (Patent No.: CN201810327904.3). The composite fitness powder disclosed in Chinese patent CN201710701559.0 uses whey protein and soy protein as the main raw materials. Its relative molecular weight is large and the solution particle size is relatively high. After entering the human digestive system, it needs to undergo step-by-step hydrolysis by multiple digestive enzymes such as pepsin and trypsin before it can be broken down into absorbable oligopeptides and amino acids. This makes it unable to meet the body's urgent needs for rapid nutrient absorption, timely energy replenishment, and repair of damaged tissues after exercise, and has certain limitations in terms of timeliness. The sports nutritional supplement disclosed in Chinese patent CN201810327904.3 contains additional free branched-chain amino acids, but the free branched-chain amino acids are easily directly decomposed during the digestion process, resulting in a significant reduction in their bioavailability, which is not conducive to the body's absorption efficiency of the nutritional powder.

[0005] The above two patents do not affect the creativity and innovation of the patent of this invention. On the contrary, they confirm that sports protein powder in the prior art generally uses high molecular weight protein or free branched-chain amino acids as raw materials, resulting in low bioavailability and insufficient absorption efficiency of the body. Therefore, the development of bioactive peptide protein powder with both low molecular weight characteristics and branched-chain amino acid enrichment advantages has become an urgent problem to be solved in the field of sports nutrition. The technical solution of the present invention can not only provide theoretical support and practical path for the research and development of sports protein powder products, but also has important significance for promoting the extension and value-added of the big health industry. Summary of the Invention

[0006] The purpose of the present invention is to provide a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids and a preparation method thereof, which solves the problems of low branched-chain amino acid content, high particle size and single raw materials in commercially available sports protein powder. The present invention has the advantages of simple process and low cost, and is suitable for factory production.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] Step 1: Egg white peptide preparation and peptide sequence determination

[0009] (1) Preparation of egg white peptide: After washing the fresh eggs, sterilize, beat the eggs, separate and collect the fresh egg whites, take an appropriate amount of egg white and mix it with water in equal proportions to prepare a 5% egg white solution. Place it in a 90℃ magnetic stirring water bath and heat it for 10 minutes to completely denature it. The denatured solution is placed in a 45℃ magnetic stirring water bath and allowed to stand for 20 minutes. Add 20,000-40,000U / g alkaline protease and hydrolyze it for 90-150 minutes. After the first step of enzymatic hydrolysis is completed, the solution temperature is raised to 55℃, and 10,000-25,000U / g flavor protease is added for the second step of enzymatic hydrolysis for 60-120 minutes. After the enzymatic hydrolysis is completed, the solution is placed in a 90℃ water bath to inactivate the enzyme for 10 minutes. After the solution temperature cools to room temperature, place it in a 4℃ centrifuge at 10,000rpm for 10 minutes, and collect the supernatant, which is the enzymatic hydrolyzate. The enzymatic hydrolysate was mixed with 50-200 mesh activated carbon at a solid-liquid ratio of 1:5-1:10 and stirred at 25-65°C for 30-120 minutes. After adsorption, the solution was filtered through a 0.22 μm filter to remove the activated carbon and then spray-dried. The inlet air temperature was 100-130°C, the outlet air temperature was 60-80°C, and the peristaltic pump was operated at 8-10 rpm. Egg white peptide was obtained after spray drying.

[0010] (2) Peptide sequence determination: The sequence of egg white peptide was analyzed using Orbitrap Elite high-field electrostatic field orbital ion hydrazine mass spectrometer.

[0011] Step 2: Preparation of sports protein powder and determination of particle size and PDI

[0012] Add 2-15 parts of egg white peptide, 3-10 parts of whey protein powder, 0.5-8 parts of skim milk powder, 0.01-0.5 parts of strawberry flavor, 0.005-0.06 parts of sucralose, 0.005-0.08 parts of complex minerals, and 0.000001-0.00007 parts of complex vitamins to a blender in a specific order and gradually mix until uniform. The product is then sieved through a 100-200 mesh sieve, packaged by weight, sealed, and stored at 4°C. The sports protein powders prepared from different raw material mixing sequences and commercially available products are added to 40-50°C water, thoroughly mixed, and their average particle size and PDI are measured using a laser particle size analyzer.

[0013] Preferably, the composite mineral is a mixture of magnesium oxide, zinc lactate and ferric pyrophosphate.

[0014] The preferred vitamin complex is vitamin C, vitamin E, vitamin A, vitamin D3, vitamin B1, vitamin B2, and vitamin B6.

[0015] Step 3: Determination of nutritional content of sports protein powder

[0016] According to the "National Food Safety Standard - Determination of Protein in Foods" (GB 5009.5-2016) and the "National Food Safety Standard - Determination of Fat in Foods" (GB 5009.6-2016) standards and using the phenol-sulfuric acid method to determine the protein, fat and carbohydrate content in the product, evaluate whether the product meets the requirements for protein supplement products in the "National Food Safety Standard - General Rules for Sports Nutrition Foods" (GB24154-2015).

[0017] Step 4: Determination of branched-chain amino acid content and sensory evaluation of sports protein powder and commercial products

[0018] The branched-chain amino acid content of the sports protein powder was determined according to the method specified in the "National Food Safety Standard - Determination of Amino Acids in Food" (GB 5009.124-2016). A sensory evaluation scoring table was developed based on relevant evaluation standards. Volunteers were recruited to conduct a sensory evaluation of the sports nutrition powder of the present invention and commercially available products to assess consumer acceptance of the sports protein powder.

[0019] The present invention has the following beneficial effects:

[0020] (1) The present invention uses egg white peptide, whey protein and skimmed milk powder as main raw materials, combined with complex minerals and complex vitamins to develop a sports nutrition food that meets the requirements of GB 24154-2015 protein supplement products.

[0021] (2) The egg white peptide containing the DKVVDVD sequence in the present invention has a high content of branched-chain amino acids. Compared with free branched-chain amino acids, egg white peptides containing branched-chain amino acids prolong the action time of amino acids through sustained release, while improving absorption efficiency, reducing the metabolic pressure caused by the first-pass effect of the liver, and reducing the risk of gastrointestinal irritation. Whey protein is a high-quality protein widely used in the field of sports nutrition food. It has a slow digestion and absorption rate and can provide a long-lasting protein supply; the complex nutrient system is designed according to the needs of sports metabolism, and enhances the maintenance of neuromuscular function and fatigue recovery efficiency. The above-mentioned raw material combination breaks through the limitations of traditional formula homogeneity, and the content of branched-chain amino acids in the product is increased by 64.42% compared with similar products on the market that are not fortified with BCAA.

[0022] (3) The present invention utilizes a specific sequence of stepwise mixing of the raw materials, resulting in a product with a smaller average particle size and lower PDI. The resulting product significantly outperforms products prepared using other mixing sequences and similar commercially available products. Furthermore, sensory evaluation scores are not significantly different from those of commercially available products, demonstrating both high absorption efficiency and superior taste. Furthermore, the preparation method of this solution is simple to operate, low-cost, and readily available raw materials make it suitable for factory production. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1Comparison of particle size measurements of the product of Example 2 and Example 3 of the present invention and commercially available sports protein powder

[0024] Figure 2 Comparison of PDI measurements of the products of Example 2 and Example 3 of the present invention and commercially available sports protein powder

[0025] Figure 3 Comparison of branched-chain amino acid content in the product of the present invention and commercially available sports protein powder

[0026] Figure 4 Sensory evaluation comparison chart of the product of the present invention and commercially available sports protein powder DETAILED DESCRIPTION

[0027] The present invention will be described below with reference to specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments.

[0028] Example 1:

[0029] The preparation of egg white peptide and peptide sequence determination include the following steps:

[0030] Fresh eggs were washed, sterilized, cracked, and separated to collect the egg white. 150g of egg white was mixed with 150g of water in equal proportions to prepare a 5% egg white solution. This solution was heated in a 90°C magnetic stirring waterbath for 10 minutes to completely denature it. The denatured solution was placed in a 45°C magnetic stirring waterbath for 20 minutes before being enzymatically digested with 30,000U / g alkaline protease for 100 minutes. After the first enzymatic hydrolysis step, the solution temperature was raised to 55°C, and 20,000U / g flavor protease was added for the second enzymatic hydrolysis step for 90 minutes. After the enzymatic hydrolysis was completed, the solution was inactivated in a 90°C waterbath. After the solution cooled to room temperature, it was centrifuged at 10,000 rpm for 10 minutes in a 4°C centrifuge. The supernatant, the enzymatic hydrolyzate, was collected. The enzymatic hydrolyzate was adsorbed on 200-mesh activated carbon at a material-to-liquid ratio of 1:10 at 35°C for 120 minutes with stirring. After adsorption, the solution was filtered through a 0.22 micron filter membrane to remove the activated carbon and then spray-dried at an air inlet temperature of 100°C, an air outlet temperature of 80°C, and a peristaltic pump of 10 rpm to obtain egg white peptide.

[0031] The egg white peptides were identified by mass spectrometry, and peptide sequences with XCorr>2.0, which are more reliable, were selected. The results are shown in Table 1. Among them, DKVVDVD, derived from egg white TWNP protein, contains 42.86% valine (V), which has a strong potential to relieve fatigue and promote muscle growth.

[0032] Table 1 Mass spectrometry results of egg white peptides

[0033]

[0034] Example 2:

[0035] The preparation of the sports protein powder and the determination of particle size and PDI in this embodiment include the following steps:

[0036] 8 parts of egg white peptide, 8 parts of whey protein powder, 6 parts of skim milk powder, 0.02 parts of strawberry flavor, 0.01 parts of sucralose, 0.005 parts of complex minerals, and 0.000001 parts of complex vitamins were added to a blender in the order of vitamins-minerals-whey protein-flavoring-skim milk powder-egg white peptide, and mixed step by step until uniform. The product was then sealed in aluminum foil bags at 20g / bag and stored at 4°C. The complex minerals were a mixture of magnesium oxide, zinc lactate, and ferric pyrophosphate. The complex vitamins were vitamin C, vitamin E, vitamin A, vitamin D3, vitamin B1, vitamin B2, and vitamin B6.

[0037] After thoroughly mixing 10g of sports nutrition powder and a commercially available product with 100mL of 40°C water, the average particle size and PDI were determined using a Nano-ZS laser particle size analyzer based on dynamic light scattering. Three replicates were performed for each sample, and the average value was calculated.

[0038] Example 3:

[0039] The preparation of the sports protein powder and the determination of particle size and PDI in this embodiment include the following steps:

[0040] 8 parts of egg white peptide, 8 parts of whey protein powder, 6 parts of skim milk powder, 0.02 parts of strawberry flavor, 0.01 parts of sucralose, 0.005 parts of complex minerals, and 0.000001 parts of complex vitamins were added to a blender in the order of egg white peptide-skim milk powder-whey protein-vitamins-minerals-flavors, and mixed step by step until uniform. The product was then packaged and sealed in aluminum foil bags at 20g / bag and stored at 4°C. The complex minerals were a mixture of magnesium oxide, zinc lactate, and ferric pyrophosphate. The complex vitamins were vitamin C, vitamin E, vitamin A, vitamin D3, vitamin B1, vitamin B2, and vitamin B6.

[0041] After thoroughly mixing 10g of sports nutrition powder and a commercially available product with 100mL of 40°C water, the average particle size and PDI were determined using a Nano-ZS laser particle size analyzer based on dynamic light scattering. Three replicates were performed for each sample, and the average value was calculated.

[0042] The particle size measurement results of the product in Example 2, the product in Example 3 and the commercially available product are as follows: Figure 1 The minimum average particle size of the solution of the product of Example 2 was 244.47 nm, which was significantly reduced by 26.76% and 28.20% compared with the product of Example 3 and the commercial product, respectively. However, there was no significant difference between the product of Example 3 and the commercial product.

[0043] The PDI test results of the product in Example 2, the product in Example 3 and the commercially available product are as follows: Figure 2 The PDI of the product in Example 2 is 0.3367, which is significantly lower than that of the product in Example 3 and the commercial product by 14.91% and 17.62%.

[0044] These results demonstrate that the present invention, through the precisely designed raw material mixing sequence, achieves a significant reduction in product particle size and effectively lowers the PDI of the solution. This means the resulting product has a larger specific surface area and a more uniform dispersion, effectively shortening the product's dissolution time and promoting full contact between the active ingredient and the absorption site, thereby significantly improving the product's absorption efficiency and significantly enhancing its bioavailability.

[0045] Example 4:

[0046] Sports protein powder was prepared as described in Example 2, and its nutritional content was determined:

[0047] (1) Protein content was determined using the Kjeldahl method according to GB 5009.5-2016: 0.5 g of sample, 0.4 g of copper sulfate, 6.0 g of potassium sulfate, and 20 mL of concentrated sulfuric acid were mixed and digested in a 420°C digestion furnace for 1 h. After cooling, the digestate was transferred to an automatic Kjeldahl nitrogen analyzer. Total nitrogen was determined by distillation, absorption, and titration, and the protein content was calculated using a coefficient of 6.25. Each sample was measured three times and the average value was taken.

[0048] (2) Determine fat content using the Soxhlet extraction method according to GB 5009.6-2016: Accurately weigh 3.000 g of sample into a constant-weight filter paper cartridge, place in a Soxhlet extractor, add 10 mL of petroleum ether, and extract under reflux in a water bath for 10 h. After extraction, remove the filter paper cartridge, evaporate the residual solvent, and weigh it. Calculate fat content by comparing the mass before and after extraction. Repeat the measurement three times for each sample and take the average value.

[0049] (3) Determination of carbohydrate content using the phenol-sulfuric acid method: Prepare a 0.0-1.0 mg / mL glucose standard solution and measure the absorbance at 490 nm to create a standard curve. Add 1.0 mL of the diluted sample to 1.0 mL of 6% phenol solution and 5.0 mL of concentrated sulfuric acid. Mix well, let stand for 30 minutes, then measure the absorbance and substitute it into the standard curve to calculate the carbohydrate content. Repeat the measurement three times for each sample and take the average value.

[0050] (4) Results and analysis

[0051] The nutritional composition of the sports protein powder produced in this project is shown in the table. The protein content is 53.36%, the fat content is 5.21%, and the carbohydrate content is 13.99%. It is worth noting that GB 24154-2015 requires that powdered protein supplements contain at least 50% protein, and this product meets this requirement.

[0052] Table 2 Determination of nutritional components of protein powder

[0053]

[0054] Example 5:

[0055] Sports protein powder was prepared as described in Example 2, and its branched-chain amino acid content and sensory evaluation were determined and compared with commercially available products. The specific determination protocol is as follows:

[0056] (1) According to GB 5009.124-2016, the content of branched-chain amino acids was determined as follows: the sample was hydrolyzed with 6 mol / L hydrochloric acid at 110°C for 24 h, the hydrolyzate was derivatized with o-phthalaldehyde-mercaptoethanol, and separated by cation exchange chromatography in an amino acid analyzer. The contents of 17 amino acids in the product were quantitatively analyzed using an ultraviolet / fluorescence detector. The content of branched-chain amino acids was calculated using the external standard method. The content of branched-chain amino acids was calculated as (L-valine content + L-isoleucine content + L-leucine content) / total free amino acid content.

[0057] (2) Sensory evaluation: The sensory standards are shown in Table 3. 20 g of the product was dissolved in 200 mL of 40°C water, and 10 professionals conducted a sensory evaluation of the product based on four dimensions: color, solubility, odor, and taste.

[0058] Table 3 Sensory evaluation standards

[0059]

[0060] (3) Results and analysis:

[0061] The content of branched-chain amino acids in the product of the present invention and commercially available products is as follows: Figure 3 The branched-chain amino acid content of similar products on the market is 13.07%, while the branched-chain amino acid content of the product of the present invention reaches 21.49%, which is significantly increased by 64.42% compared with the commercial products.

[0062] Sensory evaluation of the product of the present invention and commercially available products is as follows Figure 4 The sensory evaluation score of the commercial product was 95.27 points, while the sensory evaluation score of the product of the present invention was 96.50 points, which was not significantly different from the commercial product.

[0063] In summary, compared with commercially available products, the products of this project have no difference in sensory evaluation, but show better results in branched-chain amino acid content.

[0064] The above description is only a preferred embodiment of the present invention. Those skilled in the art may make various improvements and modifications to the above embodiment without departing from the principles and spirit of the present invention. These improvements and modifications shall fall within the scope of protection of the present invention.

Claims

1. A method for preparing a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids, characterized in that: The following steps are involved: (1) Preparation of egg white peptide: After washing fresh eggs, disinfect, beat, separate and collect fresh egg whites, take an appropriate amount of egg white and mix it with water in equal proportions to prepare a 5% egg white solution; place it in a 90°C magnetic stirring water bath and heat it for 10 minutes to completely denature it; the denatured solution is placed in a 45°C magnetic stirring water bath and allowed to stand for 20 minutes, then 20,000-40,000 U / g alkaline protease is added for enzymatic hydrolysis for 90-150 minutes; after the first step of enzymatic hydrolysis is completed, the solution temperature is raised to 55°C, and 10,000-25,000 U / g flavor protease is added for the second step of enzymatic hydrolysis for 60-120 minutes. After the enzymatic hydrolysis is completed, the solution is placed in a 90°C water bath to inactivate the enzyme for 10 minutes; after the solution temperature is cooled to room temperature, it is placed in a 4°C centrifuge at 10,000 rpm for 10 minutes, and the supernatant is collected as the enzymatic hydrolyzate; the enzymatic hydrolyzate is mixed with 50-200 mesh activated carbon at a material-liquid ratio of 1:5-1:10 at 25-65°C for 30-120 minutes; after the adsorption is completed, the solution is filtered through a 0.22 μm filter membrane to remove the activated carbon and the solution is spray-dried, with an inlet air temperature of 100-130°C, an outlet air temperature of 60-80°C, and a peristaltic pump of 8-10 rpm; after spray drying, egg white peptide is obtained; (2) Preparation of sports protein powder: 2-15 parts of egg white peptide, 3-10 parts of whey protein powder, 0.5-8 parts of skim milk powder, 0.01-0.5 parts of strawberry flavor, 0.005-0.06 parts of sucralose, 0.005-0.08 parts of complex minerals, and 0.000001-0.00007 parts of complex vitamins are added to a blender in a specific order, mixed step by step, and stirred evenly; then, the product is sieved through a 100-200 mesh sieve, packaged according to weight, sealed, and stored at 4°C; (3) The average particle size and solution dispersibility index (PDI) were determined using a laser particle size analyzer; the protein content was determined using a Kjeldahl nitrogen analyzer; the branched-chain amino acid content was determined using an amino acid analyzer; and consumer acceptance was evaluated using sensory evaluation.

2. The method for preparing a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids according to claim 1, characterized in that: The prepared egg white peptide contains the DKVVDVD sequence.

3. The method for preparing a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids according to claim 1, characterized in that: The composite mineral is a mixture of magnesium oxide, zinc lactate and ferric pyrophosphate.

4. The method for preparing a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids according to claim 1, characterized in that: The complex vitamin is a mixture of vitamin C, vitamin E, vitamin A, vitamin D3, vitamin B1, vitamin B2 and vitamin B6.

5. The method for preparing a highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids according to claim 1, characterized in that: The raw materials are mixed step by step in a specific order, and the average particle size of the obtained product is 200-300nm and the PDI is 0.2-0.

4.

6. The highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids prepared by the preparation method according to claim 1, characterized in that: The protein content accounts for 51-55%.

7. The highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids prepared by the preparation method according to claim 1, characterized in that: The content of branched-chain amino acids accounts for 19.00-23.00%.

8. The highly absorbable egg white peptide sports protein powder rich in branched-chain amino acids prepared by the preparation method according to claim 1, characterized in that: The total score of color, solubility, smell and taste in the sensory evaluation was 94.20-98.50 points.

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