Albumin active polypeptide composition for promoting healing of skin mucosa wounds

Abstract

The application discloses an albumin active polypeptide composition for promoting skin mucosa wound healing, which comprises the following components: albumin active polypeptide, yeast-beta-glucan, concentrated wolfberry juice, preservative, thickening agent, sweetening agent and water. The albumin active polypeptide is encapsulated by a carrier formed by cross-linking of phosphorylated microcrystalline cellulose and glycidyltrimethylammonium chloride modified chitosan, which can well protect the albumin active polypeptide, prevent the degradation of the active polypeptide by complex gastrointestinal digestive enzymes, effectively improve the retention rate of the active polypeptide in the gastrointestinal environment, significantly increase the enrichment amount of the active polypeptide in the small intestinal epithelial cells, promote the absorption of the active polypeptide by the small intestinal epithelial cells, and slowly release in the body, and then be transported to the target through blood circulation, so that the bioavailability of the active polypeptide is improved, and the ability of the composition for promoting skin mucosa wound healing is improved.

Description

Technical Field

[0001] This invention relates to the field of polypeptide preparation technology, and more particularly to an albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds. Background Technology

[0002] The skin is one of the organs that protects the body, helps with excretion, regulates temperature changes, and senses external signals and stimuli. It is the outermost layer of the body, connecting the body to the external environment, and is the first line of defense between the body and its environment. It is also the largest organ in the human body, accounting for 10% of total body mass, and plays a crucial role in pathogen protection, vitamin D synthesis initiation, excretion, and temperature regulation. However, skin tissue is relatively fragile and inevitably suffers varying degrees of damage in daily life, commonly including mechanical trauma, burns, scalds, scratches, postoperative wounds, and diabetic wounds.

[0003] Wound healing, or trauma healing, is the natural process of skin and epidermal tissue regeneration after injury. Normally, the epidermis and dermis of the skin exist in a balanced state, forming a protective barrier for the wound. When the eschar ruptures, the normal wound healing process begins rapidly. Its basic processes include: the hemostatic phase, the inflammatory phase, the proliferative phase, and the maturation and remodeling phase.

[0004] Wound healing is a slow process. Without intervention, local blood flow will decrease, followed by inflammatory cell infiltration, which can induce muscle spasms, seriously affecting wound healing. In severe cases, it can develop into a chronic wound. How to accelerate wound healing has become a common treatment problem worldwide.

[0005] Food-derived peptides refer to mixtures of small-molecule peptides produced from edible proteins using processes such as enzymatic hydrolysis, separation, and purification. In recent years, bioactive food-derived peptides have attracted increasing attention due to their properties such as lowering blood pressure and blood lipids, antioxidation, and antibacterial activity. Numerous studies have shown that food-derived peptides can promote skin wound healing. Compared to large-molecule proteins, small-molecule food-derived peptides are more easily absorbed by the body, and because they are derived from food, they have virtually no toxic side effects. Furthermore, when these food-derived peptides are applied to the skin, they can avoid gastrointestinal digestion and maintain their structure and biological activity. These characteristics make food-derived peptides highly valuable and promising novel therapeutic agents for accelerating skin wound healing.

[0006] CN112544988A discloses a collagen peptide composition for promoting wound healing and its preparation method. The collagen peptide composition for promoting wound healing comprises the following raw material components in parts by weight: 1-5 parts blackcurrant fruit powder, 0.2-1.5 parts compound nutritional fortifier, 70-90 parts collagen peptides, 5-20 parts whey protein isolate, 1-10 parts elderberry juice powder, and 1-12 parts prickly pear juice powder; the compound nutritional fortifier includes vitamins A, B, C, and D. The collagen peptide composition for promoting wound healing provided by this invention can provide sufficient protein, minerals, and vitamins, ensuring the rich nutrition needed for wound recovery and effectively promoting wound healing. However, this invention uses collagen peptides and whey protein isolate directly as raw materials. After ingestion, these components are easily degraded by complex digestive enzymes in the gastrointestinal tract, losing their sequence integrity and thus reducing their wound-healing effect. Summary of the Invention

[0007] In view of the above-mentioned deficiencies of the prior art, the technical problem to be solved by the present invention is to provide a food-derived albumin active polypeptide composition that has no side effects and has a good effect on promoting the healing of skin and mucous membrane wounds.

[0008] To achieve the above objectives, this invention provides an albumin-based active polypeptide composition that promotes the healing of skin and mucous membrane wounds. The inventors encapsulate the albumin-based active polypeptide with a carrier formed by cross-linking phosphorylated microcrystalline cellulose and glycidyltrimethylammonium chloride-modified chitosan. This effectively protects the albumin-based active polypeptide, preventing degradation by complex digestive enzymes in the gastrointestinal tract, effectively increasing the retention rate of the active polypeptide in the gastrointestinal environment, significantly increasing the accumulation of the active polypeptide in small intestinal epithelial cells, promoting the absorption of the active peptide by small intestinal epithelial cells, and enabling slow release in vivo. The active peptide is then transported to its target site via blood circulation, improving its bioavailability and thus enhancing the composition's ability to promote the healing of skin and mucous membrane wounds.

[0009] To achieve the above-mentioned objectives, the present invention adopts the following technical solution:

[0010] An albumin-active polypeptide composition for promoting the healing of skin and mucous membrane wounds, comprising the following components: albumin-active polypeptide, yeast-β-glucan, concentrated wolfberry juice, preservative, thickener, sweetener, and water.

[0011] Preferably, the albumin-active polypeptide composition for promoting the healing of skin and mucous membrane wounds comprises the following components, all in parts by weight: 30-50 parts albumin-active polypeptide, 10-20 parts yeast-β-glucan, 1-5 parts concentrated wolfberry juice, 1-3 parts preservative, 1-3 parts thickener, 1-3 parts sweetener, and 100-200 parts water.

[0012] Preferably, the preservative is one or a mixture of two or more of sodium citrate, sodium gluconate, potassium sorbate, and propylparaben.

[0013] Preferably, the thickener is one or a mixture of two or more of xanthan gum, pectin, and carrageenan.

[0014] Preferably, the sweetener is one or a mixture of two or more of the following: sorbitol, sodium saccharin, acesulfame potassium, cyclamate, stevia, sucralose, and erythritol.

[0015] Preferably, the albumin-active polypeptide is prepared using the following method:

[0016] 1) Disperse 20-30g of egg white powder in 50-100mL of water, heat to 80-100℃, keep warm for 5-10min, then stir at 1000-3000rpm for 2-5min; cool to 40-55℃, adjust the pH of the system to 10-11 with 0.1-0.3mol / L NaOH aqueous solution; add 1-3g of alkaline protease, and hydrolyze for 20-24h; after hydrolysis, heat to 90-95℃ and inactivate the enzyme for 5-10min; cool to 40-50℃, add 6-8g of flavor protease, and hydrolyze for 10-12h; after hydrolysis, heat to 90-95℃ and inactivate the enzyme for 5-10min, then cool to room temperature, and adjust the pH to 6-8 with 0.1-0.2mol / L hydrochloric acid to obtain the hydrolysate for later use.

[0017] 2) Add activated carbon to the enzymatic hydrolysate and let it stand for 1-2 hours; filter with gauze, centrifuge at 6000-8000 rpm for 10-15 minutes, collect the supernatant, and use 1×10⁻⁶ ppm... 4 After ultrafiltration using Da's ultrafiltration membrane, the solution was concentrated under reduced pressure to 10-30% of the original volume, and then freeze-dried at -20 to -10°C for 20-24 hours to obtain albumin peptides.

[0018] Preferably, the amount of activated carbon added in step 2) is 5-10% of the mass of the enzymatic hydrolysate.

[0019] More preferably, the albumin polypeptide is an albumin-active polypeptide encapsulated by a carrier, and its preparation method is as follows:

[0020] (1) Add 5-10g of microcrystalline cellulose to 200-300mL of 5-10wt% sodium hydroxide aqueous solution, heat to 50-80℃, and stir at 300-500rpm for 2-3h; cool and filter, collect the filtrate, add 30-50mL of 1-3wt% sodium hypochlorite aqueous solution and 5-25mL of acetic acid to the filtrate, heat to 60-80℃ and react for 2-3h, then cool to room temperature, filter, collect the filtrate, add 3-5mL of 70-80wt% phosphoric acid aqueous solution, and react at 60-80℃ for 4-6h; after the reaction is complete, cool to room temperature, filter, collect the filter cake, and wash the filter cake with water until the pH of the filtrate is 6-8 to obtain phosphorylated microcrystalline cellulose;

[0021] (2) Dissolve 3-5g of chitosan in 50-80mL of 1-5wt% acetic acid aqueous solution and heat to 40-60℃; add 10-12g of glycidyltrimethylammonium chloride, react for 10-12h, add 50-150mL of acetone, stir for 5-10min; cool to room temperature, filter, collect the filtrate, and remove the solvent under reduced pressure to obtain glycidyltrimethylammonium chloride modified chitosan;

[0022] (3) Mix 5-8g of phosphorylated microcrystalline cellulose with 30-50mL of water and sonicate at 40-60W and 150-180Hz for 10-20min; add 5-15mL of glycidyl trimethylammonium chloride modified chitosan aqueous solution at a rate of 2-3 drops / second; after the addition is complete, add 3-6g of sodium tripolyphosphate and react under light-protected conditions for 10-20min; add 0.5-3g of albumin polypeptide, stir for 15-20min and freeze-dry at -20 to -10℃ for 20-24h to obtain the carrier-encapsulated albumin active polypeptide.

[0023] Albumin, also known as albumin, is broken down by enzymes into multiple peptides composed of amino acids, known as albumin bioactive peptides. As a type of dietary polypeptide, albumin bioactive peptides have advantages such as high nutritional value, diverse biological activities, and almost no toxic side effects. However, for albumin bioactive peptides to exert their biological activity in the body, they must be degraded by complex digestive enzymes in the gastrointestinal tract, and then pass through the small intestinal epithelial absorption and osmotic barrier intact. After being fully absorbed by the body, they are transported to their target sites through blood circulation and accumulate to a certain amount before they can finally exert their regulatory physiological functions. However, albumin bioactive peptides can be cleaved by proteases on the outer edge of the brush border cell membrane of the small intestine, losing the integrity of their sequence and thus losing their physiological activity.

[0024] The inventors encapsulated albumin-active peptides using a carrier formed by cross-linking phosphorylated microcrystalline cellulose and glycidyltrimethylammonium chloride-modified chitosan, effectively protecting the active peptides. Chitosan molecules possess -OH and -NH2 groups, allowing them to bind with phosphate groups. Modification of chitosan enables it to electrostatically bind with the active peptides. The cross-linking of phosphorylated microcrystalline cellulose and glycidyltrimethylammonium chloride-modified chitosan simultaneously forms a stable and dense three-dimensional network with the active peptides, enhancing the carrier's mechanical strength, the stability of the active peptides, and their tolerance to the human gastrointestinal environment. This prevents degradation of the active peptides by complex digestive enzymes in the gastrointestinal tract, effectively increasing the retention rate of the active peptides in the gastrointestinal environment. It significantly increases the accumulation of active peptides in small intestinal epithelial cells, promoting their absorption and slow release in vivo, transporting them to the target site via blood circulation, thus improving the bioavailability of the active peptides and enhancing the composition's ability to promote the healing of skin and mucous membrane wounds.

[0025] This invention also discloses a method for preparing the albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds, comprising the following steps:

[0026] Weigh out each ingredient according to the formula, dissolve yeast-β-glucan and concentrated wolfberry juice in water, add albumin active peptides, and after ultrasonic treatment, add preservatives, thickeners and sweeteners and mix evenly to obtain an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

[0027] Preferably, the ultrasonic power is 550–800W, the ultrasonic frequency is 28–40kHz, and the ultrasonic time is 5–15min.

[0028] Compared with existing technologies, the present invention has the following advantages: The albumin-based active polypeptide composition for promoting skin and mucous membrane wound healing provided by the present invention can provide the human body with protein, vitamins, and minerals, ensuring the rich nutrition required for wound recovery and effectively promoting wound healing; by encapsulating the albumin-based active peptides with a carrier formed by cross-linking phosphorylated microcrystalline cellulose and glycidyltrimethylammonium chloride-modified chitosan, the albumin-based active peptides can be better protected, improving the stability of the active peptides, preventing the degradation of the active peptides by complex digestive enzymes in the gastrointestinal tract, effectively increasing the retention rate of the active peptides in the gastrointestinal environment, significantly increasing the enrichment of the active peptides in small intestinal epithelial cells, promoting the absorption of the active peptides by small intestinal epithelial cells, and enabling slow release in vivo, which is transported to the target site through blood circulation, thereby improving the bioavailability of the active peptides and thus improving the ability of the composition to promote skin and mucous membrane wound healing; the albumin-based active polypeptide composition for promoting skin and mucous membrane wound healing provided by the present invention has a simple preparation method, readily available raw materials, and great commercial promotion value. Detailed Implementation

[0029] To avoid unnecessary details, unless otherwise specified, all items used in the following examples are commercially available products, and all methods used are conventional methods unless otherwise specified.

[0030] The sources of some of the raw materials used in this invention are as follows:

[0031] Yeast-β-glucan, 70% content, 80 mesh particle size, moisture content ≤8%, ash content ≤3%, fat content ≤10%, protein content ≤3.5%, Shaanxi Angxu Biotechnology Co., Ltd.

[0032] Concentrated goji berry juice, specific gravity 1.2, produced by Fufeng Snow Biotechnology Co., Ltd.

[0033] Egg white powder, 99% purity, model number Y124, manufactured by Shaanxi Yuanyou Biotechnology Co., Ltd.

[0034] Alkaline protease, with an enzyme activity of 20,000 U / g, an operating temperature of 40-50℃, and an operating pH of 9-11, produced by Xingtai Wanda Bioengineering Co., Ltd.

[0035] Flavor protease, with a content of 99%, an enzyme activity of 50,000 U / g, an action temperature of 45-50℃, and an action pH of 5.5-8.5, produced by Henan Jushuo Biotechnology Co., Ltd.

[0036] Chitosan, with a degree of deacetylation of 90%, Nantong Lvshen Bioengineering Co., Ltd.

[0037] Microcrystalline cellulose, 97% purity, particle size 60-100 mesh, from Yiran Mineral Products Processing Plant, Lingshou County.

[0038] Pepsin, with an enzyme activity of 100,000 U / g, from Xi'an Musen Biotechnology Co., Ltd.

[0039] Pancreatic enzyme, with an enzyme activity of 8000 U / g, produced by Shenzhen Lefu Biotechnology Co., Ltd.

[0040] Sodium tripolyphosphate, with a content of 96% and a particle size of 0.28-0.8mm, product number SJLSN001, manufactured by Anhui Guanglan Biotechnology Co., Ltd.

[0041] Example 1

[0042] A method for preparing an albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds includes the following steps:

[0043] At a stirring speed of 1000 rpm, 200 g of yeast-β-glucan and 40 g of concentrated wolfberry juice were dissolved in 1500 g of water. Then, 500 g of albumin active peptides encapsulated in a carrier were added. After ultrasonic treatment at a power of 600 W and a frequency of 30 kHz for 10 min, 10 g of sodium citrate, 10 g of carrageenan, and 10 g of stevia were added and mixed evenly to obtain an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

[0044] The method for preparing the albumin-active polypeptide encapsulated in the carrier is as follows:

[0045] 1) Disperse 20g of egg white powder in 80mL of water, heat to 90℃, keep warm for 10min, and then stir at 2000rpm for 3min; cool to 45℃, adjust the pH of the system to 10 with 0.2mol / L NaOH aqueous solution; add 2g of alkaline protease, and hydrolyze for 24h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 8min; cool to 45℃, add 5g of flavor protease, and hydrolyze for 12h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 10min, then cool to room temperature, and adjust the pH to 7 with 0.1mol / L hydrochloric acid to obtain the hydrolysate for later use;

[0046] 2) Add activated carbon to the enzymatic hydrolysate, the amount of activated carbon added being 5% of the mass of the enzymatic hydrolysate; let stand for 2 hours; filter with gauze, centrifuge at 7000 rpm for 15 minutes, collect the supernatant, and use 1×10⁻⁶... 4 After ultrafiltration using Da's ultrafiltration membrane, the solution was concentrated under reduced pressure to 20% of the original volume, and then freeze-dried at -20°C for 24 hours to obtain albumin peptides.

[0047] 3) Add 10g of microcrystalline cellulose to 250mL of 6wt% sodium hydroxide aqueous solution, heat to 60℃, and stir at 400rpm for 3h; cool and filter, collect the filtrate, add 40mL of 2wt% sodium hypochlorite aqueous solution and 20mL of acetic acid to the filtrate, heat to 70℃ and react for 3h, cool to room temperature, filter, collect the filtrate, add 5mL of 75wt% phosphoric acid aqueous solution, and react at 70℃ for 5h; after the reaction is complete, cool to room temperature and filter, collect the filtrate cake, wash the filter cake with water until the pH of the filtrate is 7, to obtain phosphorylated microcrystalline cellulose;

[0048] 4) Dissolve 4g of chitosan in 60mL of 3wt% acetic acid aqueous solution and heat to 50℃; add 11g of glycidyltrimethylammonium chloride, react for 12h, add 100mL of acetone, stir for 10min; cool to room temperature, filter, collect the filtrate, remove the solvent under reduced pressure to obtain glycidyltrimethylammonium chloride modified chitosan.

[0049] 5) Mix 5g of phosphorylated microcrystalline cellulose with 40mL of water and sonicate at 50W and 160Hz for 15min; add 10mL of 10g / L glycidyltrimethylammonium chloride modified chitosan aqueous solution at a rate of 2 drops / second; after the addition is complete, add 4g of sodium tripolyphosphate and react for 15min under light-protected conditions; add 2.5g of albumin peptide, stir for 15min and freeze-dry at -20℃ for 24h to obtain the carrier-encapsulated albumin active peptide.

[0050] Comparative Example 1

[0051] A method for preparing an albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds includes the following steps:

[0052] At a stirring speed of 1000 rpm, 200 g of yeast-β-glucan and 40 g of concentrated wolfberry juice were dissolved in 1500 g of water. Then, 500 g of albumin active peptides encapsulated in a carrier were added. After ultrasonic treatment at a power of 600 W and a frequency of 30 kHz for 10 min, 10 g of sodium citrate, 10 g of carrageenan, and 10 g of stevia were added and mixed evenly to obtain an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

[0053] The method for preparing the albumin-active polypeptide encapsulated in the carrier is as follows:

[0054] 1) Disperse 20g of egg white powder in 80mL of water, heat to 90℃, keep warm for 10min, and then stir at 2000rpm for 3min; cool to 45℃, adjust the pH of the system to 10 with 0.2mol / L NaOH aqueous solution; add 2g of alkaline protease, and hydrolyze for 24h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 8min; cool to 45℃, add 5g of flavor protease, and hydrolyze for 12h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 10min, then cool to room temperature, and adjust the pH to 7 with 0.1mol / L hydrochloric acid to obtain the hydrolysate for later use;

[0055] 2) Add activated carbon to the enzymatic hydrolysate, the amount of activated carbon added being 5% of the mass of the enzymatic hydrolysate; let stand for 2 hours; filter with gauze, centrifuge at 7000 rpm for 15 minutes, collect the supernatant, and use 1×10⁻⁶... 4 After ultrafiltration using Da's ultrafiltration membrane, the solution was concentrated under reduced pressure to 20% of the original volume, and then freeze-dried at -20°C for 24 hours to obtain albumin peptides.

[0056] 3) Dissolve 4g of chitosan in 60mL of 3wt% acetic acid aqueous solution and heat to 50℃; add 11g of glycidyltrimethylammonium chloride, react for 12h, add 100mL of acetone, stir for 10min; cool to room temperature, filter, collect the filtrate, remove the solvent under reduced pressure to obtain glycidyltrimethylammonium chloride modified chitosan.

[0057] 4) Mix 5g of microcrystalline cellulose with 40mL of 6wt% sodium hydroxide aqueous solution and sonicate at 50W and 160Hz for 15min; add 10mL of 10g / L glycidyltrimethylammonium chloride modified chitosan aqueous solution at a rate of 2 drops / second; after the addition is complete, add 4g of sodium tripolyphosphate and react for 15min under light-protected conditions; add 2.5g of albumin peptide, stir for 15min and freeze-dry at -20℃ for 24h to obtain the carrier-encapsulated albumin active peptide.

[0058] Comparative Example 2

[0059] A method for preparing an albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds includes the following steps:

[0060] At a stirring speed of 1000 rpm, 200 g of yeast-β-glucan and 40 g of concentrated wolfberry juice were dissolved in 1500 g of water. Then, 500 g of albumin active peptides encapsulated in a carrier were added. After ultrasonic treatment at a power of 600 W and a frequency of 30 kHz for 10 min, 10 g of sodium citrate, 10 g of carrageenan, and 10 g of stevia were added and mixed evenly to obtain an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

[0061] The method for preparing the albumin-active polypeptide encapsulated in the carrier is as follows:

[0062] 1) Disperse 20g of egg white powder in 80mL of water, heat to 90℃, keep warm for 10min, and then stir at 2000rpm for 3min; cool to 45℃, adjust the pH of the system to 10 with 0.2mol / L NaOH aqueous solution; add 2g of alkaline protease, and hydrolyze for 24h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 8min; cool to 45℃, add 5g of flavor protease, and hydrolyze for 12h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 10min, then cool to room temperature, and adjust the pH to 7 with 0.1mol / L hydrochloric acid to obtain the hydrolysate for later use;

[0063] 2) Add activated carbon to the enzymatic hydrolysate, the amount of activated carbon added being 5% of the mass of the enzymatic hydrolysate; let stand for 2 hours; filter with gauze, centrifuge at 7000 rpm for 15 minutes, collect the supernatant, and use 1×10⁻⁶... 4After ultrafiltration using Da's ultrafiltration membrane, the solution was concentrated under reduced pressure to 20% of the original volume, and then freeze-dried at -20°C for 24 hours to obtain albumin peptides.

[0064] 3) Mix 5g of microcrystalline cellulose with 40mL of 6wt% sodium hydroxide aqueous solution and sonicate at 50W and 160Hz for 15min; add chitosan solution obtained by dissolving 4g of chitosan in 60mL of 3wt% acetic acid aqueous solution at a rate of 2 drops / second; after the addition is complete, add 4g of sodium tripolyphosphate and react for 15min under light-protected conditions; add 2.5g of albumin peptide, stir for 15min and freeze-dry at -20℃ for 24h to obtain carrier-encapsulated albumin active peptide.

[0065] Comparative Example 3

[0066] A method for preparing an albumin-active polypeptide composition that promotes the healing of skin and mucous membrane wounds includes the following steps:

[0067] Dissolve 200g of yeast-β-glucan and 40g of concentrated wolfberry juice in 1500g of water at a stirring speed of 1000rpm, then add 500g of albumin active peptides. After ultrasonic treatment at a power of 600W and a frequency of 30kHz for 10min, add 10g of sodium citrate, 10g of carrageenan and 10g of stevia and mix well to obtain an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

[0068] The preparation method of the albumin-active polypeptide is as follows:

[0069] 1) Disperse 20g of egg white powder in 80mL of water, heat to 90℃, keep warm for 10min, and then stir at 2000rpm for 3min; cool to 45℃, adjust the pH of the system to 10 with 0.2mol / L NaOH aqueous solution; add 2g of alkaline protease, and hydrolyze for 24h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 8min; cool to 45℃, add 5g of flavor protease, and hydrolyze for 12h; after hydrolysis, heat to 95℃ and inactivate the enzyme for 10min, then cool to room temperature, and adjust the pH to 7 with 0.1mol / L hydrochloric acid to obtain the hydrolysate for later use;

[0070] 2) Add activated carbon to the enzymatic hydrolysate, the amount of activated carbon added being 5% of the mass of the enzymatic hydrolysate; let stand for 2 hours; filter with gauze, centrifuge at 7000 rpm for 15 minutes, collect the supernatant, and use 1×10⁻⁶... 4 After ultrafiltration using Da's ultrafiltration membrane, the solution was concentrated under reduced pressure to 20% of its original volume, and then freeze-dried at -20°C for 24 hours to obtain albumin peptides.

[0071] Test Example 1

[0072] Encapsulation efficiency determination: 4 mL of albumin active peptides encapsulated with the carriers prepared in Example 1 and Comparative Examples 1-2 were placed in 10 kDa ultrafiltration centrifuge tubes and centrifuged at 4000 r / min for 10 min. The unencapsulated albumin active peptides were separated into the filtrate through the filter membrane. The absorbance of the sample at 220 nm was measured using a UV spectrophotometer. The encapsulation efficiency was calculated using the following formula:

[0073] Encapsulation efficiency = (1 - mass of free albumin-active peptides / total mass of albumin-active peptides) × 100%

[0074] The test results are shown in Table 1:

[0075] Table 1. Test results of encapsulation efficiency

[0076] Encapsulation efficiency (%) Example 1 95.2 Comparative Example 1 83.6 Comparative Example 2 78.4

[0077] As can be seen from the experimental data in Table 1, the albumin active peptides encapsulated by the carrier prepared in Example 1 have the highest encapsulation efficiency. The difference between Example 1 and other comparative examples is that the carrier used is a cross-linked chitosan modified with phosphorylated microcrystalline cellulose and glycidyl trimethylammonium chloride. The reason for this phenomenon may be that chitosan has -OH and -NH2 groups in its molecular structure, which can bind to phosphate groups. By modifying chitosan, the modified chitosan can bind to the active peptides through electrostatic attraction. The cross-linking of phosphorylated microcrystalline cellulose and chitosan modified with glycidyl trimethylammonium chloride forms a stable and dense three-dimensional network with the active peptides, which can better encapsulate the active peptides.

[0078] Test Example 2

[0079] In vitro gastrointestinal simulated digestion experiment:

[0080] Weigh out 5g of the albumin-encapsulated active peptides prepared in Examples 1 and 1-2, and the albumin-encapsulated active peptides prepared in Comparative Example 3. Add 0.1g of pepsin at 37°C, adjust the pH of the system to 2.0 with 0.1-0.5mol / L hydrochloric acid, and then stir at 120rpm for 90min for enzymatic hydrolysis. Subsequently, adjust the pH of the system to 7.0 with 1mol / L sodium hydroxide aqueous solution, add 0.2g of trypsin, and continue enzymatic hydrolysis for 240min. The component with only enzyme added and the unencapsulated albumin-encapsulated active peptides are designated as control group 1 and control group 2, respectively. After the simulated digestion, the absorbance of the remaining albumin-encapsulated active peptides at 220nm is measured using a UV spectrophotometer. The retention rate of the albumin-encapsulated active peptides before and after encapsulation is calculated using the standard curve method, as shown in the following formula:

[0081] Retention rate (%) = (mass of undegraded peptides / mass of total peptides) × 100%

[0082] The test results are shown in Table 2:

[0083] Table 2 Results of in vitro gastrointestinal simulated digestion experiments

[0084] Retention rate (%) Example 1 90.3 Comparative Example 1 82.5 Comparative Example 2 71.2 Comparative Example 3 55.8

[0085] As can be seen from the experimental data in Table 2, the albumin active peptides encapsulated by the carrier prepared in Example 1 are encapsulated by a carrier formed by cross-linking phosphorylated microcrystalline cellulose and glycidyl trimethylammonium chloride-modified chitosan. The chitosan modified with glycidyl trimethylammonium chloride can bind to the active peptides through electrostatic binding. The cross-linking of phosphorylated microcrystalline cellulose and glycidyl trimethylammonium chloride-modified chitosan forms a stable and dense three-dimensional network with the active peptides, which improves the mechanical strength of the carrier and effectively prevents the degradation of active peptides by pepsin and pancreatic enzymes in the gastrointestinal tract.

[0086] Test Example 3

[0087] Mouse wound healing test:

[0088] Fifty mice, weighing 18-22g, half male and half female, were randomly divided into five groups: a blank control group, a group treated with an albumin-active polypeptide composition that promotes skin and mucous membrane wound healing, and a group treated with the albumin-active polypeptide composition that promotes skin and mucous membrane wound healing. A 1.5×1.5cm wound was created on the neck and back of each mouse to establish the wound model. After model establishment, the wound was rinsed with physiological saline, disinfected with iodine and alcohol, covered with petroleum jelly gauze, and then wrapped with sterile dry gauze. The group treated with the albumin-active polypeptide composition that promotes skin and mucous membrane wound healing was fed the albumin-active polypeptide composition prepared in Example 1 and Comparative Examples 1-3 at a dose of 1.5g / kg body weight. The control group was given the same volume of physiological saline. The treatments were administered by gavage once daily for 9 consecutive days. Each mouse's wound was rinsed with physiological saline daily and then treated using the same method. After 9 days, the wound area was measured, and the wound healing rate was calculated. The formula for calculating the wound healing rate is as follows:

[0089] Wound healing rate = (Original wound area - Unhealed wound area) / Original wound area

[0090] The test results are averaged and are shown in Table 3.

[0091] Table 3 Results of mouse wound healing test

[0092] wound healing rate Example 1 0.62±0.03 Comparative Example 1 0.55±0.02 Comparative Example 2 0.49±0.01 Comparative Example 3 0.41±0.02 Blank group 0.35±0.03

[0093] As can be seen from the data in Table 3, the albumin active polypeptide composition for promoting skin and mucous membrane wound healing prepared in Example 1 has the best wound healing effect. The possible reason is that after the albumin active polypeptide is encapsulated by the carrier formed by cross-linking phosphorylated microcrystalline cellulose and glycidyl trimethylammonium chloride modified chitosan, it prevents the degradation of the active polypeptide by the complex digestive enzyme system in the gastrointestinal tract, effectively improves the retention rate of the active polypeptide in the gastrointestinal environment, significantly increases the enrichment of the active polypeptide in the small intestinal epithelial cells, promotes the absorption of the active peptide by the small intestinal epithelial cells, and can be slowly released in vivo and transported to the target site through blood circulation, thereby improving the bioavailability of the active peptide and thus improving the ability of the composition to promote skin and mucous membrane wound healing.

Claims

1. An albumin-based active polypeptide composition for promoting the healing of skin and mucous membrane wounds, characterized in that, It includes the following components, by weight: 30-50 parts albumin active peptides, 10-20 parts yeast-β-glucan, 1-5 parts concentrated wolfberry juice, 1-3 parts preservatives, 1-3 parts thickeners, 1-3 parts sweeteners, and 100-200 parts water. The albumin-active polypeptide is a carrier-encapsulated albumin-active polypeptide, and its preparation method is as follows: (1) Add 5-10g of microcrystalline cellulose to 200-300mL of 5-10wt% sodium hydroxide aqueous solution, heat to 50-80℃, and stir at 300-500rpm for 2-3h; cool and filter, collect the filtrate, add 30-50mL of 1-3wt% sodium hypochlorite aqueous solution and 5-25mL of acetic acid to the filtrate, heat to 60-80℃ and react for 2-3h, then cool to room temperature, filter, collect the filtrate, add 3-5mL of 70-80wt% phosphoric acid aqueous solution, and react at 60-80℃ for 4-6h; after the reaction is complete, cool to room temperature, filter, collect the filter cake, and wash the filter cake with water until the pH of the filtrate is 6-8 to obtain phosphorylated microcrystalline cellulose; (2) Dissolve 3-5g of chitosan in 50-80mL of 1-5wt% acetic acid aqueous solution and heat to 40-60℃; add 10-12g of glycidyltrimethylammonium chloride and react for 10-12h, then add 50-150mL of acetone and stir for 5-10min; cool to room temperature, filter, collect the filtrate, and remove the solvent under reduced pressure to obtain glycidyltrimethylammonium chloride modified chitosan; (3) Mix 5-8g of phosphorylated microcrystalline cellulose with 30-50mL of water and sonicate at 40-60W and 150-180Hz for 10-20min; add 5-15mL of glycidyl trimethylammonium chloride modified chitosan aqueous solution at a rate of 2-3 drops / second; after the addition is complete, add 3-6g of sodium tripolyphosphate and react under light-protected conditions for 10-20min; add 0.5-3g of albumin polypeptide, stir for 15-20min and freeze-dry at -20~-10℃ for 20-24h to obtain the carrier-encapsulated albumin active polypeptide.

2. The albumin-active polypeptide composition for promoting skin and mucous membrane wound healing as described in claim 1, characterized in that: The preservative is one or a mixture of two or more of sodium citrate, sodium gluconate, potassium sorbate, and propylparaben.

3. The albumin-active polypeptide composition for promoting skin and mucous membrane wound healing as described in claim 1, characterized in that: The thickener is one or a mixture of two or more of xanthan gum, pectin, and carrageenan.

4. The albumin-active polypeptide composition for promoting skin and mucous membrane wound healing as described in claim 1, characterized in that: The sweetener is one or a mixture of two or more of the following: sorbitol, sodium saccharin, acesulfame potassium, cyclamate, stevia, sucralose, and erythritol.

5. The method for preparing the albumin-active polypeptide composition for promoting skin and mucous membrane wound healing as described in any one of claims 1-4, characterized in that, The process includes the following steps: dissolving yeast-β-glucan and concentrated wolfberry juice in water, adding albumin active peptides, and then treating with ultrasound. Finally, adding preservatives, thickeners, and sweeteners and mixing thoroughly yields an albumin active peptide composition that promotes the healing of skin and mucous membrane wounds.

6. The method for preparing the albumin-active polypeptide composition for promoting skin and mucous membrane wound healing as described in claim 5, characterized in that: The ultrasonic power is 550~800W, the ultrasonic frequency is 28~40kHz, and the ultrasonic time is 5~15min.