A composition containing an elastin peptide having anti-wrinkle firming efficacy and a preparation method thereof

Abstract

The present application relates to the field of food, especially to a composition containing elastin peptide with anti-wrinkle and firming effect and a preparation method. The composition with anti-wrinkle and firming effect is composed of 15-25 parts by weight of elastin-collagen binding peptide powder, 8-12 parts by weight of red beet juice powder, 4-6 parts by weight of sambucus williamsii fruit juice powder, 1-4 parts by weight of sturgeon oligopeptide powder, 1-4 parts by weight of tremella fuciformis extract and 15-20 parts by weight of resistant dextrin. The present application can produce significant synergistic effect by compounding each component in the formula according to a certain content, effectively improve the content of elastin and collagen in the skin, increase the skin elasticity, effectively reduce the wrinkles and delay the skin aging.

Description

Technical Field

[0001] This invention relates to the food industry, and more particularly to a composition containing elastin peptides that has anti-wrinkle and firming effects, and a method for its preparation. Background Technology

[0002] As consumers increasingly demand anti-aging effects from skincare products, elastin and collagen, as key ingredients for maintaining skin elasticity and firmness, have become core active substances in anti-wrinkle cosmetics and functional foods. Elastin peptides and collagen peptides, due to their small molecular weight and high bioavailability, can effectively promote the synthesis of elastin and collagen in the skin through oral or topical application, thereby improving aging problems such as wrinkles and sagging.

[0003] Existing compositions often directly combine elastin peptides and collagen peptides. However, due to the lack of directional cross-linking of peptide segments, the target sites of elastin peptides and collagen peptides are dispersed, making it difficult to simultaneously activate the synthesis pathways of elastin and collagen through a synergistic mechanism, thus limiting the anti-wrinkle effect.

[0004] In addition, this study found that red beets contain a large number of nutrients, such as betaine, anthocyanins, vitamins and minerals, which can effectively work with elastin peptides and collagen peptides to synthesize skin proteins, thereby increasing skin elasticity and effectively reducing wrinkles. However, the geosmin contained in red beets can cause an off-flavor when the combination is taken. Traditional high-temperature inactivation or adsorption methods to remove geosmin will destroy the heat-sensitive nutrients in it, resulting in a reduction in the anti-aging effect of red beets. Summary of the Invention

[0005] To address the aforementioned technical deficiencies, this invention presents a method for preparing a composition containing elastin peptides with anti-wrinkle and firming effects. This method cross-links elastin peptides and collagen peptides to enhance their synergistic effect. Furthermore, it effectively deodorizes red beets by preparing enzyme-fixed magnetic beads. The resulting composition is odorless and can effectively increase skin elasticity, reduce wrinkles, and delay skin aging.

[0006] A composition containing elastin peptides with anti-wrinkle and firming effects comprises the following ingredients: 15-25 parts by weight of elastin-collagen-binding peptide powder, 8-12 parts by weight of red beet juice powder, 4-6 parts by weight of elderberry juice powder, 1-4 parts by weight of fish maw oligopeptide powder, 1-4 parts by weight of Pleurotus ostreatus extract and 15-20 parts by weight of resistant dextrin.

[0007] The elastic-collagen-binding peptide powder is made by purifying a mixture of bovine neck ligament and cod skin through alkali treatment and acid treatment, then sequentially hydrolyzing it with collagenase, flavor protease and transglutaminase, and finally separating, freeze-drying and pulverizing it.

[0008] Red beet juice powder is made by juicing red beets, then enzymatically hydrolyzing it using enzyme-immobilized magnetic beads assembled from neodymium iron boron magnetic beads treated with laccase, β-cyclodextrin, glutaraldehyde, and 3-aminopropyltriethoxysilane, followed by freeze-drying and pulverizing.

[0009] A method for preparing a composition containing elastin peptides with anti-wrinkle and firming effects includes the following steps:

[0010] S1: Preparation of elastic-collagen-binding peptide powder;

[0011] S2: Preparation of enzyme-immobilized magnetic beads;

[0012] S3: Deodorization of red beet juice using magnetic ball method and compounding with protein-bound peptide powder.

[0013] Furthermore, the preparation of the elastic-collagen-binding peptide powder in step S1 specifically includes the following steps:

[0014] S1.1: Clean the bovine neck ligament and cod skin at a mass ratio of 1:(1-1.5), then remove the surface fat and muscle, and grind them together in a meat grinder. Add a 0.1-0.15mol / L NaOH solution at a material-to-liquid ratio of 1:(10-12)g / mL. Refrigerate and stir at 3-4℃ and 120-150rpm for 10-12 hours. Then filter and wash the solid with water until neutral. After draining the water, add deionized water at a material-to-liquid ratio of 1:(8-10)g / mL, and add hydrochloric acid to adjust the pH to 2-3. Shake in a water bath at a frequency of 200-250 times / minute for 30-40 minutes. Filter and wash the solid with water until neutral. Then add deionized water at a material-to-liquid ratio of 1:(10-15)g / mL. Stir in a water bath at 60-65℃ for 6-8 hours. Discard the supernatant and filter to obtain the pretreated composite protein.

[0015] S1.2: Mix the pretreated composite protein and deionized water at a material-to-liquid ratio of 1:(10-15) g / mL. Add 0.1-0.15wt% collagenase and hydrolyze at 50-55℃ for 2-2.5 hours. Then adjust the pH to 8-8.5, add 0.1-0.15wt% flavor protease, maintain the temperature and hydrolyze for 2-2.5 hours. Then inactivate the enzyme at 105℃ for 8-10 minutes. Add deionized water to maintain the original volume. Then adjust the pH to 6-6.5, maintain the temperature at 50-52℃, add 0.05-0.1wt% transglutaminase and stir evenly. Keep warm for 2-3 hours. After inactivating the enzyme at 105℃ for 8-10 minutes, place it in a centrifuge and centrifuge at 8000-10000rpm for 10-15 minutes. Take the supernatant and freeze-dry it. After pulverizing, obtain elastic-collagen-binding peptide powder.

[0016] Further, step S2, preparing enzyme-immobilized magnetic beads, specifically includes the following steps:

[0017] S2.1: Mix laccase and β-cyclodextrin in a mass ratio of 1:(4-5) in a container and stir magnetically for 25-30 minutes at 3-4℃ and 200-220rpm to obtain the enzyme-β-CD complex.

[0018] S2.2: Disperse NdFeB magnetic beads ultrasonically in anhydrous toluene, then add 2-5% by volume of 3-aminopropyltriethoxysilane, heat to 70-80℃ under a nitrogen atmosphere, stir for 8-12 hours, then wash with toluene 2-3 times, and vacuum dry at 60-65℃ to obtain aminated magnetic beads.

[0019] S2.3: Mix the aminated magnetic beads and the enzyme-β-CD complex at a mass ratio of 1:(2.5-3) in a container, slowly inject 6-6.5wt% glutaraldehyde over 25-30 minutes, and then carry out a shaking reaction at 23-25℃ and 150-180rpm for 2-2.5 hours. Collect the solid by magnetic separation, wash it 2-3 times with phosphate buffer, air dry it under a nitrogen atmosphere, and store it at 3-4℃ to obtain enzyme-immobilized magnetic beads.

[0020] Furthermore, step S3, the magnetic ball deodorization of red beet juice and the compounding of protein-bound peptide powder, includes the following steps:

[0021] S3.1: Wash fresh, non-rotten red beets, peel them, cut them into pieces, and juice them. Then, pass the juice through an 80-100 mesh sieve to obtain red beet juice. Add 2-3 wt% of enzyme-fixed magnetic beads, and stir at a constant temperature of 25-27℃ and 100-120 rpm for 1-2 hours. Then, separate the enzyme-fixed magnetic beads using a 0.5-0.6T magnetic field. Filter the remaining liquid through a filter membrane to obtain deodorized red beet juice. Freeze-dry the juice at -40--30℃ and then pulverize it to obtain red beet juice powder.

[0022] S3.2: Mix 15-25 parts by weight of elastic-collagen-binding peptide powder, 8-12 parts by weight of red beet juice powder, 4-6 parts by weight of elderberry juice powder, 1-4 parts by weight of fish maw oligopeptide powder, 1-4 parts by weight of golden top pleurotus extract and 15-20 parts by weight of resistant dextrin evenly, sterilize and seal packaging to obtain a composition with anti-wrinkle and firming effects.

[0023] Furthermore, in step S1.1, when adding NaOH solution for alkaline treatment, the NaOH solution should be replaced after 5-6 hours.

[0024] Furthermore, in step S2.2, the particle size of the neodymium iron boron magnetic beads is 0.3-0.5 mm.

[0025] Furthermore, the pore size of the filter membrane in step S3.1 is 0.22-0.25 μm.

[0026] The beneficial effects are as follows: 1. This invention produces an elastic-collagen-binding peptide powder by compound enzymatic hydrolysis of bovine cervical ligament and cod skin. The powder contains elastin peptides and collagen peptides. After consumption, it can promote the expression of related proteins and factors in the elastin and collagen synthesis pathway, such as lysyl oxidase and transforming growth factor, and promote the synthesis of elastin and collagen in the skin. The natural betaine and anthocyanins in red beet juice powder can also inhibit skin aging. In addition, vitamins and a large number of minerals can also synergistically promote the synthesis of elastin and collagen. The various components in the formula are compounded in a certain amount to produce a significant synergistic effect, effectively increase the content of elastin and collagen in the skin, increase skin elasticity, effectively reduce wrinkles, and delay skin aging.

[0027] 2. This invention involves purifying bovine cervical ligament and cod skin, then sequentially hydrolyzing them with collagenase, flavor protease, and transglutaminase. First, collagenase specifically decomposes the proteins, followed by enzymatic hydrolysis with flavor protease, which has a broad action site on proteins, yielding highly active elastin peptides and collagen peptides. After inactivating the flavor protease and collagenase, transglutaminase treatment enables the elastin peptides and collagen peptides to pair and bind, allowing them to simultaneously act on the elastin and collagen synthesis pathways, thereby enhancing their synergistic effect and further promoting the synthesis of elastin and collagen in the skin.

[0028] 3. This invention first amides neodymium iron boron magnetic beads, and then, under the action of glutaraldehyde crosslinking agent, immobilizes the enzyme-β-CD complex prepared by laccase and β-cyclodextrin on the surface of the aminated magnetic beads to obtain enzyme-immobilized magnetic beads. Using enzyme-immobilized magnetic beads loaded with laccase to enzymatically hydrolyze red beet juice, it can catalytically oxidize geosmin in red beet juice, converting phenolic hydroxyl groups into quinones, which are further polymerized into large molecular odorless substances. Moreover, the enzyme-immobilized magnetic beads can be separated by magnetic field, quickly stopping the enzymatic hydrolysis process without the need for high-temperature inactivation, thus avoiding the destruction of nutrients by high temperature. Attached Figure Description

[0029] Figure 1 This is a scanning electron microscope image of the elastic-collagen binding peptide prepared in Example 1 of the present invention, magnified 500 times. Detailed Implementation

[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0031] Example 1: A composition containing elastin peptides with anti-wrinkle and firming effects and its preparation method, comprising the following steps:

[0032] S1: Preparation of elastic-collagen-binding peptide powder

[0033] S1.1: Clean the beef neck ligament and cod skin at a 1:1 mass ratio, remove the surface fat and muscle, and grind them together in a meat grinder. Add a 0.1mol / L NaOH solution at a material-to-liquid ratio of 1:10g / mL. Refrigerate and stir at 3℃ and 120rpm for 10 hours. Replace the NaOH solution after 5 hours. Filter the solid and wash it with water until neutral. Drain the water and add deionized water at a material-to-liquid ratio of 1:8g / mL. Add hydrochloric acid to adjust the pH to 2. Shake in a water bath at 200 times / minute for 30 minutes. Filter the solid and wash it with water until neutral. Add deionized water at a material-to-liquid ratio of 1:10g / mL. Stir in a water bath at 60℃ for 6 hours. Discard the supernatant and filter to obtain the pretreated composite protein.

[0034] S1.2: Mix the pretreated composite protein and deionized water at a material-to-liquid ratio of 1:10 g / mL. Add 0.1 wt% collagenase and hydrolyze at 50℃ for 2 hours. Then adjust the pH to 8, add 0.1 wt% flavor protease, maintain the temperature for 2 hours, and then inactivate the enzyme at 105℃ for 8 minutes. Add deionized water to maintain the original volume, then adjust the pH to 6, maintain the temperature at 50℃, add 0.05 wt% transglutaminase, stir well, and incubate for 2 hours. After enzyme inactivation at 105℃ for 8 minutes, the sample was centrifuged at 8000 rpm for 10 minutes. The supernatant was then freeze-dried to obtain elastin-collagen-binding peptides. Small pieces of the elastin-collagen-binding peptides were fixed onto a slide. The peptides were then torn to create small cross-sections and subjected to vacuum gold sputtering. The images were then observed and photographed under a scanning electron microscope at 500x magnification, resulting in a 500x magnified image of the elastin-collagen-binding peptides. Figure 1 As shown, the remaining elastin-collagen-binding peptides were pulverized to obtain elastin-collagen-binding peptide powder.

[0035] S2: Preparation of enzyme-immobilized magnetic beads

[0036] S2.1: Laccase and β-cyclodextrin were mixed in a mass ratio of 1:4 and placed in a container. The mixture was magnetically stirred for 25 minutes at 3°C ​​and 200 rpm to obtain the enzyme-β-CD complex.

[0037] S2.2: Neodymium iron boron magnetic beads with a particle size of 0.3 mm were ultrasonically dispersed in anhydrous toluene, and then 2% by volume of 3-aminopropyltriethoxysilane was added. The mixture was heated to 70°C under a nitrogen atmosphere and stirred for 8 hours. Then it was washed twice with toluene and dried under vacuum at 60°C to obtain aminated magnetic beads.

[0038] S2.3: Aminated magnetic beads and enzyme-β-CD complex were mixed in a mass ratio of 1:2.5 and placed in a container. 6 wt% glutaraldehyde was slowly injected over 25 minutes. The mixture was then subjected to a shaking reaction at 23°C and 150 rpm for 2 hours. The solid was collected by magnetic separation, washed twice with phosphate buffer, air-dried under a nitrogen atmosphere, and stored at 3°C ​​to obtain enzyme-immobilized magnetic beads.

[0039] S3: Magnetic ball deodorization of red beet juice and compounding with protein-bound peptide powder

[0040] S3.1: Wash fresh, non-rotten red beets, peel them, cut them into pieces, and juice them. Then, pass the juice through an 80-mesh sieve to obtain red beet juice. Add 2wt% enzyme-fixed magnetic beads and stir at a constant temperature of 25℃ and 100rpm for 1 hour. Then, separate the enzyme-fixed magnetic beads using a 0.5T magnetic field. Filter the remaining liquid through a 0.22μm pore size filter membrane to obtain deodorized red beet juice. Freeze-dry the juice at -40℃ and then pulverize it to obtain red beet juice powder.

[0041] S3.2: Mix 15 parts by weight of elastic-collagen-binding peptide powder, 8 parts by weight of red beet juice powder, 4 parts by weight of elderberry juice powder, 1 part by weight of fish maw oligopeptide powder, 1 part by weight of golden top pleurotus extract and 15 parts by weight of resistant dextrin evenly, sterilize and seal the mixture to obtain a composition with anti-wrinkle and firming effects.

[0042] Example 2: A composition containing elastin peptides with anti-wrinkle and firming effects and its preparation method, comprising the following steps:

[0043] S1: Elastic Collagen-Binding Peptide Powder

[0044] S1.1: Clean the beef neck ligament and cod skin at a mass ratio of 1:1.5, then remove the surface fat and muscle, and grind them together in a meat grinder. Add a 0.1mol / L NaOH solution at a material-to-liquid ratio of 1:10g / mL. Refrigerate and stir at 3℃ and 120rpm for 10 hours. Replace the NaOH solution after 5 hours. Filter the solid and wash it with water until neutral. Drain the water and add deionized water at a material-to-liquid ratio of 1:8g / mL. Add hydrochloric acid to adjust the pH to 2. Shake in a water bath at a frequency of 200 times / minute for 30 minutes. Filter the solid and wash it with water until neutral. Add deionized water at a material-to-liquid ratio of 1:10g / mL. Stir in a water bath at 60℃ for 6 hours. Discard the supernatant and filter to obtain the pretreated composite protein.

[0045] S1.2: Mix the pretreated composite protein and deionized water at a material-to-liquid ratio of 1:10 g / mL. Add 0.15 wt% collagenase and hydrolyze at 50℃ for 2 hours. Then adjust the pH to 8, add 0.15 wt% flavor protease, maintain the temperature and hydrolyze for 2 hours. Then inactivate the enzyme at 105℃ for 8 minutes. Add deionized water to maintain the original volume. Then adjust the pH to 6, maintain the temperature at 50℃, add 0.1 wt% transglutaminase and stir well. Keep warm for 2 hours. After inactivating the enzyme at 105℃ for 8 minutes, place it in a centrifuge and centrifuge at 8000 rpm for 10 minutes. Take the supernatant and freeze-dry it. After pulverizing, obtain elastic-collagen-binding peptide powder.

[0046] S2: Preparation of enzyme-immobilized magnetic beads

[0047] S2.1: Laccase and β-cyclodextrin were mixed in a mass ratio of 1:5 and placed in a container. The mixture was magnetically stirred for 25 minutes at 3°C ​​and 200 rpm to obtain the enzyme-β-CD complex.

[0048] S2.2: Neodymium iron boron magnetic beads with a particle size of 0.3 mm were ultrasonically dispersed in anhydrous toluene, and then 5% by volume of 3-aminopropyltriethoxysilane was added. The mixture was heated to 70°C under a nitrogen atmosphere and stirred for 8 hours. Then it was washed twice with toluene and dried under vacuum at 60°C to obtain aminated magnetic beads.

[0049] S2.3: Aminated magnetic beads and enzyme-β-CD complex were mixed in a 1:3 mass ratio and placed in a container. 6.5wt% glutaraldehyde was slowly injected over 25 minutes. The mixture was then subjected to a shaking reaction at 23°C and 150 rpm for 2 hours. The solid was collected by magnetic separation, washed twice with phosphate buffer, air-dried under a nitrogen atmosphere, and stored at 3°C ​​to obtain enzyme-immobilized magnetic beads.

[0050] S3: Magnetic ball deodorization of red beet juice and compounding with protein-bound peptide powder

[0051] S3.1: Wash fresh, non-rotten red beets, peel them, cut them into pieces, and juice them. Then, pass the juice through an 80-mesh sieve to obtain red beet juice. Add 3wt% enzyme-fixed magnetic beads and stir at a constant temperature of 25℃ and 100rpm for 1 hour. Then, separate the enzyme-fixed magnetic beads using a 0.5T magnetic field. Filter the remaining liquid through a 0.22μm pore size filter membrane to obtain deodorized red beet juice. Freeze-dry the juice at -40℃ and then pulverize it to obtain red beet juice powder.

[0052] S3.2: Mix 25 parts by weight of elastic-collagen-binding peptide powder, 12 parts by weight of red beet juice powder, 6 parts by weight of elderberry juice powder, 4 parts by weight of fish maw oligopeptide powder, 4 parts by weight of golden top pleurotus extract and 20 parts by weight of resistant dextrin evenly, sterilize and seal the mixture to obtain a composition with anti-wrinkle and firming effects.

[0053] Example 3: A composition containing elastin peptides with anti-wrinkle and firming effects and its preparation method, comprising the following steps:

[0054] S1: Elastic Collagen-Binding Peptide Powder

[0055] S1.1: Clean the beef neck ligament and cod skin at a 1:1 mass ratio, then remove the surface fat and muscle, and grind them together in a meat grinder. Add a 0.15mol / L NaOH solution at a material-to-liquid ratio of 1:12g / mL. Refrigerate and stir at 4℃ and 150rpm for 12 hours. Replace the NaOH solution after 6 hours. Filter the solid and wash it with water until neutral. Drain the water and add deionized water at a material-to-liquid ratio of 1:10g / mL. Add hydrochloric acid to adjust the pH to 3. Shake in a water bath at 250 times / minute for 40 minutes. Filter the solid and wash it with water until neutral. Add deionized water at a material-to-liquid ratio of 1:15g / mL. Stir in a water bath at 65℃ for 8 hours. Discard the supernatant and filter to obtain the pretreated composite protein.

[0056] S1.2: Mix the pretreated composite protein and deionized water at a material-to-liquid ratio of 1:15 g / mL. Add 0.1 wt% collagenase and hydrolyze at 55℃ for 2.5 hours. Then adjust the pH to 8.5, add 0.1 wt% flavor protease, maintain the temperature and hydrolyze for 2.5 hours. Then inactivate the enzyme at 105℃ for 10 minutes. Add deionized water to maintain the original volume. Then adjust the pH to 6.5, the temperature to 52℃, add 0.05 wt% transglutaminase and stir well. Keep warm for 3 hours. After inactivating the enzyme at 105℃ for 10 minutes, place it in a centrifuge and centrifuge at 10000 rpm for 15 minutes. Take the supernatant and freeze-dry it. After pulverizing, obtain elastic-collagen-binding peptide powder.

[0057] S2: Preparation of enzyme-immobilized magnetic beads

[0058] S2.1: Laccase and β-cyclodextrin were mixed in a mass ratio of 1:4 and placed in a container. The mixture was magnetically stirred for 30 minutes at 4°C and 220 rpm to obtain the enzyme-β-CD complex.

[0059] S2.2: Neodymium iron boron magnetic beads with a particle size of 0.5 mm were ultrasonically dispersed in anhydrous toluene, and then 2% by volume of 3-aminopropyltriethoxysilane was added. The mixture was heated to 80°C under a nitrogen atmosphere and stirred for 12 hours. Then it was washed three times with toluene and dried under vacuum at 65°C to obtain aminated magnetic beads.

[0060] S2.3: Aminated magnetic beads and enzyme-β-CD complex were mixed in a 1:3 mass ratio and placed in a container. 6 wt% glutaraldehyde was slowly injected over 30 minutes. The mixture was then subjected to a shaking reaction at 25°C and 180 rpm for 2.5 hours. The solid was collected by magnetic separation, washed three times with phosphate buffer, air-dried under a nitrogen atmosphere, and stored at 4°C to obtain enzyme-immobilized magnetic beads.

[0061] S3: Magnetic ball deodorization of red beet juice and compounding with protein-bound peptide powder

[0062] S3.1: Wash fresh, non-rotten red beets, peel them, cut them into pieces, and juice them. Then, pass the juice through a 100-mesh sieve to obtain red beet juice. Add 2wt% enzyme-fixed magnetic beads and stir at a constant temperature of 27℃ and 120rpm for 2 hours. Then, separate the enzyme-fixed magnetic beads using a 0.6T magnetic field. Filter the remaining liquid through a 0.22μm pore size filter membrane to obtain deodorized red beet juice. Freeze-dry the juice at -30℃ and then pulverize it to obtain red beet juice powder.

[0063] S3.2: Mix 15 parts by weight of elastic-collagen-binding peptide powder, 8 parts by weight of red beet juice powder, 4 parts by weight of elderberry juice powder, 1 part by weight of fish maw oligopeptide powder, 1 part by weight of golden top pleurotus extract and 15 parts by weight of resistant dextrin evenly, sterilize and seal the mixture to obtain a composition with anti-wrinkle and firming effects.

[0064] Comparative Example 1: The difference from Example 1 is that the composition with anti-wrinkle and firming effects prepared in Comparative Example 1 is formulated with 15 parts by weight of elastic-collagen-binding peptide powder, 4 parts by weight of elderberry juice powder, 1 part by weight of fish maw oligopeptide powder, 1 part by weight of Pleurotus ostreatus extract and 15 parts by weight of resistant dextrin.

[0065] Comparative Example 2: The difference from Example 1 is that the composition with anti-wrinkle and firming effects prepared in Comparative Example 2 is formulated with 15 parts by weight of elastic-collagen-binding peptide powder, 8 parts by weight of red beet juice powder, 1 part by weight of fish maw oligopeptide powder, 1 part by weight of Pleurotus ostreatus extract and 15 parts by weight of resistant dextrin.

[0066] Comparative Example 3: The difference from Example 1 is that the composition with anti-wrinkle and firming effects prepared in Comparative Example 3 is formulated with 15 parts by weight of elastic-collagen-binding peptide powder, 8 parts by weight of red beet juice powder, 4 parts by weight of elderberry juice powder, 1 part by weight of Pleurotus ostreatus extract and 15 parts by weight of resistant dextrin.

[0067] Comparative Example 4: The difference from Example 1 is that the composition with anti-wrinkle and firming effects prepared in Comparative Example 4 is formulated with 15 parts by weight of elastic-collagen-binding peptide powder, 8 parts by weight of red beet juice powder, 4 parts by weight of elderberry juice powder, 1 part by weight of fish maw oligopeptide powder and 15 parts by weight of resistant dextrin.

[0068] Comparative Example 5: The difference from Example 1 is that in Comparative Example 5, transglutaminase was not added in step S1.2 for 2 hours of heat preservation treatment. The remaining steps were the same as in Example 1 to obtain elastic-collagen complex peptide powder. The subsequent elastic-collagen bound peptide powder was replaced with an equal mass of elastic-collagen complex peptide powder.

[0069] Comparative Example 6: The difference from Example 1 is that Comparative Example 6 did not use enzyme-immobilized magnetic beads to treat red beet juice, but the remaining steps were the same as in Example 1.

[0070] Experiment 1: The compositions prepared in Examples 1-3 and the compositions prepared in Comparative Examples 1-5 were all prepared by adding 5 wt% white sugar to obtain samples. Fifty-four SD rats, half male and half female, weighing 200±20g, were divided into 9 groups of 6 rats each. One group served as the blank group, and the remaining groups served as experimental groups. The rats were administered one of the samples prepared in Examples 1-3 or Comparative Examples 1-5 by gavage. The dosage was 10mg / kg based on the elastic-collagen-binding peptide powder. The administration was once a day for 30 days.

[0071] On days 1 and 31 of the experiment, after cleaning the back skin of the same area of ​​mice in each group by removing the fine hairs, skin elasticity was tested using a skin elasticity tester (Cutometer dualMPA580). The average value of each group was recorded as R1 and R2, respectively. The rate of change of R is calculated as (R1-R2) / R1×100%. The closer the skin elasticity R value is to 1, the better the skin elasticity. The higher the rate of change of R, the better the effect of the composition with anti-wrinkle and firming effects on improving skin elasticity. The recorded rates of change are shown in Table 1.

[0072] After elasticity testing, mice were euthanized by cervical dislocation. 200 mg of skin tissue was accurately weighed from each mouse in each group. The skin tissue from each group was mixed and minced to obtain meat tissue. Three 200 mg portions of meat tissue were randomly selected from different mice in each group. The meat tissue was processed according to GB / T9695.23-2008, and the hydroxyproline content was measured and calculated. The collagen content was then calculated as hydroxyproline content / 13% (hydroxyproline accounts for approximately 13% of the total amino acid content of collagen). The average value was taken. The collagen content of the control group was set to 100%. The proportions of Examples 1-3 and Comparative Examples 1-5 relative to the control group were calculated. The test results are shown in Table 1.

[0073] Table 1

[0074]

[0075] As can be seen from the data in Table 1, compared with the blank group, the anti-wrinkle and firming composition prepared in the examples can significantly increase the collagen content in mouse skin and improve the elasticity of mouse skin. This proves that the composition prepared from elasticity-collagen-binding peptide powder, red beet juice powder, elderberry juice powder, fish maw oligopeptide powder, and Pleurotus ostreatus extract can effectively increase the collagen content in the skin, reduce wrinkles, and improve skin elasticity. Furthermore, as can be seen from the data in Comparative Examples 1-5, when one of the components is removed, the collagen content and elasticity in mouse skin decrease significantly, proving that there is a synergistic effect among the components of the composition provided in this example, which can provide anti-wrinkle and firming effects for the skin.

[0076] Experiment 2: Red beet juice powder prepared in equal masses from Examples 1-3 and Comparative Example 6 was used as samples. The content of geosmin in the samples was determined by gas chromatography-mass spectrometry. Three parallel tests were performed, and then the odor was evaluated. The test results are shown in Table 2.

[0077] Table 2

[0078]

[0079] As can be seen from Table 2, the content of geosmin in the red beet juice powder prepared in Examples 1-3 is significantly lower than that in Comparative Example 6. This proves that the present application can significantly reduce the content of geosmin by preparing enzyme-immobilized magnetic beads to catalytically oxidize geosmin in red beet juice, and the resulting red beet juice powder has no odor.

[0080] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A composition containing elastin peptides with anti-wrinkle and firming effects, characterized in that, It includes the following ingredients: 15-25 parts by weight of elastin-collagen-binding peptide powder, 8-12 parts by weight of red beet juice powder, 4-6 parts by weight of elderberry juice powder, 1-4 parts by weight of fish maw oligopeptide powder, 1-4 parts by weight of golden top pleurotus extract and 15-20 parts by weight of resistant dextrin. The elastic-collagen-binding peptide powder is made by purifying a mixture of bovine neck ligament and cod skin through alkali treatment and acid treatment, then sequentially hydrolyzing it with collagenase, flavor protease and transglutaminase, and finally separating, freeze-drying and pulverizing it. Red beet juice powder is made by juicing red beets, then enzymatically hydrolyzing it using enzyme-immobilized magnetic beads assembled from neodymium iron boron magnetic beads treated with laccase, β-cyclodextrin, glutaraldehyde and 3-aminopropyltriethoxysilane, and then freeze-drying and pulverizing it. The method for preparing the composition containing elastin peptides and having anti-wrinkle and firming effects includes the following steps: S1: Preparation of elastic-collagen-binding peptide powder S1.1: Clean the bovine neck ligament and cod skin at a mass ratio of 1:(1-1.5), then remove the surface fat and muscle, and grind them together in a meat grinder. Add a 0.1-0.15mol / L NaOH solution at a material-to-liquid ratio of 1:(10-12)g / mL. Refrigerate and stir at 3-4℃ and 120-150rpm for 10-12 hours. Then filter and wash the solid with water until neutral. After draining the water, add deionized water at a material-to-liquid ratio of 1:(8-10)g / mL, and add hydrochloric acid to adjust the pH to 2-3. Shake in a water bath at a frequency of 200-250 times / minute for 30-40 minutes. Filter and wash the solid with water until neutral. Then add deionized water at a material-to-liquid ratio of 1:(10-15)g / mL. Stir in a water bath at 60-65℃ for 6-8 hours. Discard the supernatant and filter to obtain the pretreated composite protein. S1.2: Mix the pretreated composite protein and deionized water at a ratio of 1:(10-15) g / mL until homogeneous. Add 0.1-0.15wt% collagenase and hydrolyze at 50-55℃ for 2-2.5 hours. Then adjust the pH to 8-8.5, add 0.1-0.15wt% flavor protease, and hydrolyze at the same temperature for 2-2.5 hours. Then inactivate the enzyme at 105℃ for 8-10 minutes. Add deionized water to maintain the original volume. Then adjust the pH to 6-6.5, maintain the temperature at 50-52℃, add 0.05-0.1wt% transglutaminase, stir until homogeneous, keep warm for 2-3 hours, inactivate the enzyme at 105℃ for 8-10 minutes, and then place in a centrifuge. Centrifuge at 8000-10000 rpm for 10-15 minutes. Take the supernatant, freeze dry, and pulverize to obtain elastic-collagen-binding peptide powder. S2: Preparation of enzyme-immobilized magnetic beads S2.1: Mix laccase and β-cyclodextrin in a mass ratio of 1:(4-5) in a container and stir magnetically for 25-30 minutes at 3-4℃ and 200-220rpm to obtain the enzyme-β-CD complex. S2.2: Disperse NdFeB magnetic beads ultrasonically in anhydrous toluene, then add 2-5% by volume of 3-aminopropyltriethoxysilane, heat to 70-80℃ under a nitrogen atmosphere, stir for 8-12 hours, then wash with toluene 2-3 times, and vacuum dry at 60-65℃ to obtain aminated magnetic beads. S2.3: Mix the aminated magnetic beads and the enzyme-β-CD complex at a mass ratio of 1:(2.5-3) in a container, slowly inject 6-6.5wt% glutaraldehyde over 25-30 minutes, and then carry out a shaking reaction at 23-25℃ and 150-180rpm for 2-2.5 hours. Collect the solid by magnetic separation, wash it 2-3 times with phosphate buffer, air dry it under a nitrogen atmosphere, and store it at 3-4℃ to obtain enzyme-immobilized magnetic beads. S3: Magnetic ball deodorization of red beet juice and compounding with protein-bound peptide powder S3.1: Wash fresh, non-rotten red beets, peel them, cut them into pieces, and juice them. Then, pass the juice through an 80-100 mesh sieve to obtain red beet juice. Add 2-3 wt% of enzyme-fixed magnetic beads, and stir at a constant temperature of 25-27℃ and 100-120 rpm for 1-2 hours. Then, separate the enzyme-fixed magnetic beads using a 0.5-0.6T magnetic field. Filter the remaining liquid through a filter membrane to obtain deodorized red beet juice. Freeze-dry the juice at -40--30℃ and then pulverize it to obtain red beet juice powder. S3.2: Mix 15-25 parts by weight of elastic-collagen-binding peptide powder, 8-12 parts by weight of red beet juice powder, 4-6 parts by weight of elderberry juice powder, 1-4 parts by weight of fish maw oligopeptide powder, 1-4 parts by weight of golden top pleurotus extract and 15-20 parts by weight of resistant dextrin evenly, sterilize and seal packaging to obtain a composition with anti-wrinkle and firming effects.

2. The composition containing elastin peptides with anti-wrinkle and firming effects as described in claim 1, characterized in that, In the preparation method of the composition containing elastin peptides with anti-wrinkle and firming effects, when adding NaOH solution for alkaline treatment in step S1.1, the NaOH solution is replaced after 5-6 hours.

3. The composition containing elastin peptides with anti-wrinkle and firming effects as described in claim 1, characterized in that, In the preparation method of the composition containing elastin peptides with anti-wrinkle and firming effects, the particle size of the neodymium iron boron magnetic beads in step S2.2 is 0.3-0.5 mm.

4. The composition containing elastin peptides with anti-wrinkle and firming effects as described in claim 1, characterized in that, In the preparation method of the composition containing elastin peptides with anti-wrinkle and firming effects, the pore size of the filter membrane in step S3.1 is 0.22-0.25 μm.

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