Composite peptide nutritional composition for promoting wound healing and preparation process and application thereof

Abstract

The invention belongs to the technical field of medical nutritional food, and particularly relates to a composite peptide nutritional composition for promoting wound healing and a preparation process and application thereof. The composite peptide nutritional composition for promoting wound healing is prepared from the following preparation raw materials in percentage by weight: 15%-25% of fish collagen peptide, 0.03%-0.05% of cattle spleen peptide, 0.10%-0.30% of blood protein oligopeptide, 0.10%-0.30% of albumin peptide, 0.01%-0.03% of ginseng powder, 15%-20% of a composite fermentation extract, 10%-20% of a plant extract, 20%-25% of prebiotics and the balance of whey protein matrix, totaling 100%. The self-made fish collagen peptide, the compound fermentation extract and other raw materials are adopted, the content of various active ingredients such as healing-promoting peptide fragments and anti-inflammatory repair is increased, the bioavailability of the plant extract is improved through the embedding technology, and therefore the wound healing rate and quality are remarkably improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical nutritional foods, and in particular relates to a composite peptide nutritional composition for promoting wound healing, and a preparation process and application thereof. Background Art

[0002] In the field of wound repair and tissue regeneration medicine, accelerating wound healing and improving healing quality through nutritional interventions have always been a key focus of clinical research. Traditional treatments often rely on exogenous growth factors or antibiotics, but these are limited by low bioavailability, the risk of drug resistance, and a single mechanism of action. In recent years, complex peptide nutritional compositions that combine natural active ingredients with biotechnology have become a research hotspot, but existing technologies still have limitations.

[0003] As a key functional ingredient, collagen peptides are susceptible to damage by conventional enzymatic hydrolysis, resulting in insufficient release of short peptide segments with healing activity, which directly affects the migration ability of fibroblasts. Although some products contain active ingredients, they are single-ingredient, and while they can improve wound healing, the improvement is limited and the healing efficiency is low. Furthermore, as an oral nutritional product, sensitive active ingredients must be protected from environmental damage such as gastric acid, ensuring that they have high bioavailability so that they can be absorbed by the human body and exert their efficacy.

[0004] Therefore, there is an urgent need to develop a complex peptide nutritional composition that can promote wound healing through oral administration, improve the speed and quality of wound healing by enriching the content of active short peptide segments and effective ingredients, and provide an efficient solution for postoperative wounds, trauma, ulcers, dermatitis and other wounds. Summary of the Invention

[0005] The purpose of the present invention is to provide a composite peptide nutritional composition, preparation process, and application for promoting wound healing, which significantly improves the wound healing rate and quality, and provides an efficient solution for wounds such as postoperative wounds, trauma, ulcers, and dermatitis.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A composite peptide nutritional composition for promoting wound healing. The raw materials for its preparation, calculated by weight percentage, include 15%-25% fish collagen peptide, 0.03%-0.05% bovine spleen peptide, 0.10%-0.30% blood protein oligopeptide, 0.10%-0.30% albumin peptide, 0.01%-0.03% ginseng powder, 15%-20% composite fermentation extract, 10%-20% plant extract, 20%-25% prebiotics, and whey protein matrix supplemented with the remainder to 100%.

[0008] The method for preparing fish collagen peptide comprises the following steps: A1, mixing collagen peptide raw material with sodium citrate solution, centrifuging and then washing with water to obtain demineralized fish scales;

[0009] A2. Add water to the demineralized fish scales and adjust the pH, add alkaline protease, react, add flavor protease to react, inactivate the enzyme, ultrafilter using an ultrafiltration membrane, and then freeze-dry.

[0010] Preferably, the method for preparing the fish collagen peptide comprises the following steps: A1, mixing the collagen peptide raw material with a 0.1M sodium citrate solution, stirring at 0-4°C for 10-14h, centrifuging and washing with water until neutral, to obtain demineralized fish scales;

[0011] A2. Add water to the demineralized fish scales and adjust the pH to 7. Add alkaline protease, react at 25°C for 1-3 hours, add flavor protease, react at 25°C for 1-3 hours, inactivate the enzyme, first use an ultrafiltration membrane with a molecular weight cutoff of 3000Da to obtain an ultrafiltrate, then use an ultrafiltration membrane with a molecular weight cutoff of 300Da to obtain a retained material, and the retained material is freeze-dried.

[0012] Preferably, the collagen peptide raw material includes fish scales or fish skin.

[0013] Preferably, the fish scales include tilapia scales; and the fish skin includes deep-sea cod skin.

[0014] Preferably, the solid-liquid ratio of the collagen peptide raw material to the 0.1M sodium citrate solution is 1 g: (8-12) mL.

[0015] Preferably, the mass ratio of the demineralized fish scales to water is 1:(4-6).

[0016] Preferably, the added amount of the alkaline protease is 300-400 U / g collagen peptide raw material.

[0017] Preferably, the added amount of the flavor protease is 300-400 U / g collagen peptide raw material.

[0018] Preferably, the enzyme inactivation conditions are: temperature of 85-90° C. and time of 10-15 min.

[0019] Preferably, the freeze-drying conditions are: pre-freezing at -40°C for 4 hours, and then drying at a temperature of -30°C and a pressure of 8-12 Pa for 24 hours.

[0020] By first demineralizing the collagen peptide raw material with sodium citrate and then preparing fish collagen peptides through a two-stage enzymatic hydrolysis, the product's ability to promote wound healing can be enhanced. This may be because decalcification with sodium citrate at low temperature can protect the triple helix structure. Then, through a step-by-step enzymatic hydrolysis technique, alkaline protease first cleaves the hydrophobic peptide bonds in the non-helical region of the collagen, initially releasing large molecular weight peptides. Subsequently, flavor protease is added for deep hydrolysis at the same temperature, precisely cleaving specific sites in the helical region and releasing pro-healing functional peptides such as Gly-Pro-Hyp tripeptide. Finally, by controlling the molecular weight cut-off of the ultrafiltration membrane, functional peptides with a concentrated molecular weight are obtained. This ensures the biological activity of the collagen peptides while promoting intestinal absorption, thereby increasing the collagen peptide's ability to promote fibroblast migration and accelerate wound healing. The step-by-step enzymatic hydrolysis avoids mutual inhibition of enzyme activity due to overlapping substrate specificity, ensuring that each enzyme acts independently under optimal conditions, significantly improving the degree of hydrolysis and product yield. Considering cost, tilapia scales are generally more economical to use as a raw material. However, deep-sea cod skin is richer in higher-quality collagen, amino acids, and various essential trace elements, and has higher bioactivity. It can effectively promote skin cell regeneration and repair, and is often used in high-end products. In actual industry, the choice between tilapia scales and deep-sea cod skin can be determined according to needs.

[0021] Preferably, the preparation method of the composite fermentation extract comprises the following steps: inoculating yeast on culture medium 1, obtaining fermentation product 1 after fermentation, adding Cordyceps mycelium to obtain culture medium 2, inoculating Cordyceps militaris and Cordyceps militaris, obtaining fermentation product 2 after culture, adding Centella asiatica powder to obtain culture medium 3, inoculating mold, fermenting to obtain fermentation product 3, inoculating Lactobacillus plantarum, after fermentation, membrane filtration, concentration, and spray drying to obtain.

[0022] Preferably, the preparation method of the composite fermentation extract comprises the following steps: inoculating yeast on culture medium 1, shaking and fermenting at 28-32°C and 120-180rpm for 72 hours to obtain fermentation product 1, adding Cordyceps mycelium to obtain culture medium 2, inoculating Cordyceps militaris and Cordyceps militaris, and culturing at 23-25°C for 10 days to obtain fermentation product 2, adding Centella asiatica powder to obtain culture medium 3, inoculating mold, shaking and fermenting at 28-30°C and 180-220rpm for 96 hours to obtain fermentation product 3, inoculating Lactobacillus plantarum, and anaerobic fermentation at 37°C for 48 hours, filtering with a ceramic membrane with a molecular weight cutoff of 50kDa, vacuum concentrating at 45°C to a solid content of ≥30%, and spray drying to obtain the product.

[0023] Preferably, the yeast comprises Candida utilis.

[0024] Preferably, the inoculation amount of the yeast is 1×10 7 CFU / g culture medium 1.

[0025] Preferably, the culture medium 1 comprises, by weight percentage, 5% soybean meal, 3%-4% sucrose, 0.01% sodium selenite, and water to make up the balance to 100%.

[0026] Preferably, the added amount of the Cordyceps mycelium is consistent with the mass of the fermentation product 1.

[0027] Preferably, the inoculation amount of the cicada fungus and Cordyceps militaris is the same, both of which are 2×10 7 -3×10 7 CFU / g culture medium 2.

[0028] Preferably, the added amount of the Centella asiatica powder is 2%-4% of the mass of the fermentation product 2.

[0029] Preferably, the mold includes Monascus and Aspergillus niger.

[0030] Preferably, the inoculation amount of Monascus is 1×10 7 -2×10 7 CFU / g culture medium 3; the inoculum size of Aspergillus niger was 2×10 6 -4×10 6 CFU / g culture medium 3.

[0031] Preferably, the inoculation amount of the plant lactobacillus is 4×10 6 -6×10 6 CFU / g fermentation product 3.

[0032] Preferably, the vacuum degree of the vacuum concentration is 0.08-0.1 MPa.

[0033] Preferably, the specific conditions of the spray drying are: air inlet temperature of 150-220°C, air outlet temperature of 80-100°C, and atomizer speed of 20,000-30,000 rpm.

[0034] By preparing a composite fermentation extract, not only can the immune system and anti-inflammatory effects of the prepared composite peptide nutritional composition be improved, but it can also promote tissue repair and regulate intestinal flora, shortening the healing time through a multi-party synergistic effect. This may be because first, soybean meal is used as a nitrogen source, sucrose is used as a carbon source, and sodium selenite is used as a selenium source. The biotransformation of selenium is achieved through fermentation by Candida utilis, generating organic selenium forms such as selenomethionine, while also producing glycolic acid that can soften the cell wall of the subsequent Cordyceps mycelium and provide a pretreatment substrate for subsequent fermentation. The fermentation liquid of stage 1 is then mixed with the Cordyceps mycelium and inoculated with cicada fungus and Cordyceps militaris. The cicada fungus secretes laccase to degrade the chitin skeleton of the mycelium, and Cordyceps militaris produces a cordycepin precursor. The two work synergistically to increase the cordycepin content in the product. Centella asiatica powder was then added, and the mixture was inoculated with Monascus purpureus and Aspergillus niger. Monascus purpureus secretes red pigments to inhibit bacterial contaminants, while Aspergillus niger produces saccharifying enzymes to activate triterpenoid saponins. The two synergistically enhance the conversion rate of triterpenoid compounds. The resulting asiaticoside also produces a synergistic antioxidant effect with salidroside in the Rhodiola rosea extract. Finally, Lactobacillus plantarum was inoculated for anaerobic fermentation. This lactic acid bacteria metabolizes short-chain fatty acids, lowering the pH of the system, promoting the dissolution of active ingredients, and increasing the γ-aminobutyric acid (GABA) content in the final product, significantly enhancing the anti-inflammatory activity of the composition. The resulting composite fermentation extract is rich in active ingredients such as cordycepin, selenomethionine, asiaticoside, GABA, and short-chain fatty acids. These synergistic effects can shorten the inflammatory phase of wounds, promote fibroblast proliferation and collagen synthesis, increase the rate of granulation tissue formation, inhibit excessive scar formation, and accelerate wound healing.

[0035] By selecting the progressive fermentation method, the product of the previous fermentation process is used as the substrate of the next fermentation process. Compared with fermentation alone, the generation of active ingredients can be further improved, while improving the conversion efficiency of plant lactobacillus, thereby increasing the content of active ingredients in the final product, and improving the wound healing effect and efficiency of the composite peptide nutritional composition. This may be because the fermentation process of the present invention has a metabolite cascade amplification effect on the one hand, and the seleno-isoflavones of stage 1 can induce stage 2 cicada fungus to produce cordycepin, and the cordycepin of stage 2 can suppress the risk of bacterial contamination of stage 3 Monascus, thereby improving the fungal reaction efficiency, and plant lactobacillus can degrade the by-products such as phytic acid produced by Monascus fermentation. On the other hand, in the coupled fermentation of stage 3 Monascus and Aspergillus niger, the cellulase produced by Aspergillus niger can release the triterpenoid saponins in the cell wall of Centella asiatica, thereby improving the product repair healing effect. In addition, the progressive fermentation liquid contains rich amino acids and short-chain fatty acids, which can provide a carbon source for stage 4 plant lactobacillus, promote its bioconversion efficiency, and improve the content of various active ingredients in the composite fermentation extract. Moreover, the progressive fermentation method avoids the complicated process of preparing multiple active ingredients and the separation and purification in each step, thus reducing costs.

[0036] Preferably, the plant extract includes green tea extract and rhodiola rosea extract.

[0037] Preferably, the mass ratio of the green tea extract to the rhodiola rosea extract is (3-5):1.

[0038] Preferably, the preparation method of the green tea extract comprises the following steps:

[0039] B1. Grind dried green tea leaves through a 60-mesh sieve to obtain green tea powder, add 65%-75% ethanol aqueous solution, perform ultrasonic-assisted extraction, filter, and concentrate by rotary evaporation at 45°C to a concentration of 1-3 times the mass of the green tea powder to obtain a concentrate;

[0040] B2. Take the concentrated liquid, wall material liquid and emulsifier, mix them, homogenize them twice, freeze-dry them, and grind them through a 100-mesh sieve to obtain the product.

[0041] Preferably, the solid-to-liquid ratio of the green tea powder to the ethanol aqueous solution with a mass fraction of 65%-75% is 1 g: (14-16) mL.

[0042] Preferably, the specific conditions of the ultrasound-assisted extraction are: frequency of 38-42 kHz, power of 220-270 W, time of 25-35 min, and temperature of 25-35°C.

[0043] Preferably, the components of the wall material liquid include, by weight percentage of the wall material liquid, 2% sodium alginate, 0.5% pectin, and water as the balance to 100%.

[0044] Preferably, the emulsifier is lecithin, and the added amount is 0.1% of the mass of the concentrate.

[0045] Preferably, the volume ratio of the concentrated liquid to the wall material liquid is 1:3.

[0046] Preferably, the homogenization pressure is 75-85 MPa, and the homogenization time is 3-5 min.

[0047] Preferably, the freeze-drying conditions are: pre-freezing at -40°C for 2 hours, and then drying at a temperature of -30°C and a pressure of 8-12 Pa for 24 hours.

[0048] Preferably, the content of salidroside in the Rhodiola rosea extract is ≥3%.

[0049] In some preferred embodiments, the Rhodiola rosea extract comes from Xi'an Tianyi Biotechnology Co., Ltd.

[0050] Nanoemulsification of green tea extract using a sodium alginate-pectin composite wall material improves its oral bioavailability, resists gastric acidity, and enhances the retention of active ingredients such as EGCG, thereby enhancing bioavailability. This is likely because EGCG, an active ingredient in green tea extract, is less stable than salidroside, an active ingredient in rhodiola rosea extract. It is susceptible to light, heat, oxidation, and degradation in gastric acid, making it less effective when administered orally. Therefore, emulsification is necessary. Sodium alginate and pectin form a double-layer wall material, stabilizing the microcapsule structure through electrostatic bonding. Lecithin reduces interfacial tension, enabling high-pressure homogenization to achieve nanoscale emulsification. Freeze-drying preserves the integrity of the microcapsules, enabling controlled release of active ingredients such as EGCG, prolonging their retention at the wound site, enhancing antioxidant and anti-inflammatory effects, and synergizing with salidroside to reduce collagen degradation.

[0051] Preferably, the prebiotics include resistant dextrin and galacto-oligosaccharide.

[0052] Preferably, the mass ratio of the resistant dextrin to galacto-oligosaccharide is 1:(1-2); more preferably, it is 2:3.

[0053] Preferably, the whey protein matrix comprises, by weight percentage of the whey protein matrix, 5%-10% lactose, 1%-2% vitamin C, and whey protein powder as the balance to 100%.

[0054] Preferably, the vitamin C is L-ascorbic acid.

[0055] In some preferred embodiments, an appropriate amount of vitamins, including but not limited to vitamin B1, vitamin B2, and vitamin B6, may be added to the complex peptide nutritional composition of the present invention, with the added amount being 0.01%-0.02% of the mass of the complex peptide nutritional composition.

[0056] The second aspect of the present invention provides a process for preparing the composite peptide nutritional composition for promoting wound healing, comprising the following steps:

[0057] S1, granulating and crushing fish collagen peptide, bovine spleen peptide, blood protein oligopeptide, albumin peptide, prebiotics and whey protein matrix to obtain a mixture;

[0058] S2. Add ginseng powder, compound fermentation extract, and plant extract to the mixture in sequence, and mix them to obtain the product.

[0059] Preferably, the preparation process comprises:

[0060] S1. Dry-mixing fish collagen peptide, bovine spleen peptide, blood protein oligopeptide, albumin peptide, prebiotics and whey protein matrix in a mixer, crushing to a particle size of ≤40 mesh to obtain a mixture;

[0061] S2. Add ginseng powder, compound fermentation extract, and plant extract to the mixture in sequence and mix under nitrogen protection to obtain the product.

[0062] In step S1, the specific conditions for dry mixing granulation are: rotation speed of 15-25 rpm, mixing time of 5-15 min, roller pressure of 60-70 kN, and roller spacing of 0.8-1.2 mm.

[0063] In step S2, the specific conditions for mixing are: a rotation speed of 5-15 rpm and a mixing time of 10-20 min.

[0064] The third aspect of the present invention provides the application of the composite peptide nutritional composition for promoting wound healing, which is applied in the field of medical nutritional food, including postoperative wounds, trauma, ulcers, dermatitis, etc.

[0065] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0066] 1. The present invention provides a composite peptide nutritional composition for promoting wound healing. By using homemade fish collagen peptides, composite fermentation extracts and other raw materials, the content of various active ingredients such as pro-healing peptides and anti-inflammatory repair is increased, and the bioavailability of plant extracts is improved through encapsulation technology, thereby significantly improving the wound healing rate and quality. In particular, it has excellent healing effects on iatrogenic wounds caused by tumor radiotherapy and chemotherapy, as well as pressure sores, diabetic foot, burns and other difficult-to-heal wounds, providing an efficient solution for postoperative wounds, trauma, ulcers, dermatitis and other wounds.

[0067] 2. The present invention first demineralizes fish scales / fish skin with sodium citrate and then uses a two-stage enzymatic hydrolysis to prepare fish collagen peptides, thereby improving the product's ability to promote wound healing.

[0068] 3. The present invention prepares a composite fermentation extract, which can not only improve the immune system and anti-inflammatory effects of the prepared composite peptide nutritional composition, but also promote tissue repair and regulate intestinal flora, thereby shortening the healing time through multi-party synergistic effects.

[0069] 4. The present invention adopts a progressive fermentation method and uses the product of the previous fermentation process as the substrate of the next fermentation process. Compared with fermentation alone, it can further improve the production of active ingredients and at the same time improve the conversion efficiency of Lactobacillus plantarum, thereby increasing the content of active ingredients in the final product and improving the wound healing effect and efficiency of the complex peptide nutritional composition.

[0070] 5. The present invention uses a sodium alginate-pectin composite wall material to nano-emulsify and embed the green tea extract, thereby improving its oral bioavailability, resisting gastric acid, and increasing the retention rate of active ingredients such as EGCG, thereby improving bioavailability. DETAILED DESCRIPTION

[0071] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0072] The raw materials used in the present invention are all commercially available, specifically:

[0073] Bovine spleen peptide, blood protein oligopeptide, albumin peptide, and ginseng powder are all from Xi'an Zebang Biotechnology Co., Ltd.

[0074] Alkaline protease, with an enzyme activity of approximately 500,000 U / g, was obtained from Shanghai Huashang Xiangyang Biotechnology Co., Ltd.

[0075] Flavor protease, with an activity of approximately 30,000 U / g, was obtained from Nanning Dongheng Huadao Biotechnology Co., Ltd.

[0076] Soybean meal, from Shandong Rundong Agricultural Technology Co., Ltd.

[0077] Candida utilis, strain number CCTCC CY 20081274; Monascus purpureus, strain number CCTCC HF2008672; Aspergillus niger, strain number CCTCC AF 2022057; Lactobacillus plantarum, strain number CCTCC AB 2022408; all were obtained from China Center for Type Culture Collection.

[0078] Cicada fungus, strain number cfcc 50943; Cordyceps militaris, strain number cfcc 88944; both are from the China Forestry Microbial Culture Collection Management Center.

[0079] Cordyceps mycelium comes from Fufeng Snow Biotechnology Co., Ltd.

[0080] Sodium alginate, pectin, and lecithin were all from Shanghai MacLean Biochemical Technology Co., Ltd.

[0081] Rhodiola rosea extract, salidroside content ≥3%, from Xi'an Tianyi Biotechnology Co., Ltd.

[0082] Resistant dextrin, from Roquette, France, NUTRIOSE® FB06.

[0083] Galacto-oligosaccharide was obtained from Shanghai MacLean Biochemical Technology Co., Ltd.

[0084] Lactose was obtained from Jiangsu Caiwei Biotechnology Co., Ltd.

[0085] Whey protein powder, whey protein concentrate, WPC80, from Shandong Jingyun Food Ingredients Co., Ltd.

[0086] Example 1

[0087] The present embodiment provides a composite peptide nutritional composition for promoting wound healing. The raw materials for preparing the composite peptide nutritional composition include, by weight percentage, 20% fish collagen peptide, 0.04% bovine spleen peptide, 0.20% blood protein oligopeptide, 0.20% albumin peptide, 0.02% ginseng powder, 18% composite fermentation extract, 15% plant extract, 22% prebiotics, and whey protein matrix supplemented with the remainder to 100%.

[0088] The preparation method of the fish collagen peptide comprises the following steps:

[0089] A1. Mix the collagen peptide raw material with 0.1M sodium citrate solution, stir at 4°C for 12 hours, centrifuge, and wash with water until neutral to obtain demineralized fish scales;

[0090] A2. Water was added to the demineralized fish scales and the pH was adjusted to 7. Alkaline protease was added and the reaction was carried out at 25°C for 2 hours. Flavor protease was then added and the reaction was carried out at 25°C for 2 hours. The enzyme was inactivated. An ultrafiltration membrane with a molecular weight cutoff of 3000 Da was used to obtain an ultrafiltrate, which was then treated with an ultrafiltration membrane with a molecular weight cutoff of 300 Da to obtain a retained material. The obtained material was freeze-dried.

[0091] The collagen peptide raw material is deep-sea cod skin.

[0092] The solid-liquid ratio of the collagen peptide raw material to the 0.1M sodium citrate solution is 1 g:10 mL.

[0093] The mass ratio of the demineralized fish scales to water is 1:5.

[0094] The added amount of the alkaline protease is 350 U / g collagen peptide raw material.

[0095] The added amount of the flavor protease is 350 U / g collagen peptide raw material.

[0096] The enzyme inactivation conditions are: temperature 85° C., time 15 min.

[0097] The freeze-drying conditions are: pre-freezing at -40°C for 4 hours, and then drying at a temperature of -30°C and a pressure of 10 Pa for 24 hours.

[0098] The preparation method of the composite fermentation extract comprises the following steps: inoculating yeast on a culture medium 1, performing shaking fermentation at 30°C and 150 rpm for 72 hours to obtain a fermentation product 1, adding cordyceps mycelium to obtain a culture medium 2, inoculating cicadae and cordyceps militaris, and statically culturing at 23°C for 10 days to obtain a fermentation product 2, adding Centella asiatica powder to obtain a culture medium 3, inoculating mold, performing shaking fermentation at 28°C and 200 rpm for 96 hours to obtain a fermentation product 3, inoculating Lactobacillus plantarum, performing anaerobically fermenting at 37°C for 48 hours, filtering through a ceramic membrane with a molecular weight cutoff of 50 kDa, vacuum concentrating at 45°C to a solid content of 30%, and spray drying to obtain the product.

[0099] The yeast is Candida utilis.

[0100] The inoculation amount of the yeast was 1×10 7 CFU / g culture medium 1.

[0101] The culture medium 1 comprises, by weight percentage, 5% soybean meal, 2.5% sucrose, 0.01% sodium selenite, and water to make up the balance to 100%.

[0102] The amount of the Cordyceps mycelium added is consistent with the mass of the fermentation product 1.

[0103] The inoculation amount of the cicada fungus and Cordyceps militaris was the same, both of which were 2.5×10 7 CFU / g culture medium 2.

[0104] The added amount of the Centella asiatica powder is 3% of the mass of the fermentation product 2.

[0105] The molds include Monascus and Aspergillus niger.

[0106] The inoculum size of Monascus was 1.5×10 7 CFU / g culture medium 3; the inoculum size of Aspergillus niger was 3×10 6 CFU / g culture medium 3.

[0107] The inoculation amount of the plant lactobacillus was 5×10 6 CFU / g fermentation product 3.

[0108] The vacuum degree of the vacuum concentration is 0.09 MPa.

[0109] The specific conditions of the spray drying are: air inlet temperature of 180° C., air outlet temperature of 90° C., and atomizer speed of 25,000 rpm.

[0110] The plant extracts are green tea extract and rhodiola rosea extract, with a mass ratio of 4:1.

[0111] The preparation method of the green tea extract comprises the following steps:

[0112] B1. Grind dried green tea leaves through a 60-mesh sieve to obtain green tea powder. Add 70% ethanol aqueous solution by mass, perform ultrasonic-assisted extraction, filter, and concentrate by rotary evaporation at 45°C to a concentration twice the mass of the green tea powder to obtain a concentrate.

[0113] B2. Take the concentrated liquid, wall material liquid and emulsifier, mix them, homogenize them twice, freeze-dry them, and grind them through a 100-mesh sieve to obtain the product.

[0114] The solid-to-liquid ratio of the green tea powder to the 70% by mass ethanol aqueous solution is 1 g:15 mL.

[0115] The specific conditions of the ultrasound-assisted extraction are: frequency of 40 kHz, power of 250 W, time of 30 min, and temperature of 30°C.

[0116] The components of the wall material liquid, calculated by weight percentage of the wall material liquid, include 2% sodium alginate, 0.5% pectin, and water in an amount that is supplemented to 100%.

[0117] The emulsifier is lecithin, and the added amount is 0.1% of the mass of the concentrated liquid.

[0118] The volume ratio of the concentrated liquid to the wall material liquid is 1:3.

[0119] The homogenization pressure is 85 MPa and the homogenization time is 4 min.

[0120] The freeze-drying conditions are: pre-freezing at -40°C for 2 hours, and then drying at a temperature of -30°C and a pressure of 10 Pa for 24 hours.

[0121] The prebiotics are resistant dextrin and galacto-oligosaccharide, with a mass ratio of 2:3.

[0122] The whey protein matrix comprises, by weight percentage of the whey protein matrix, 8% lactose, 1.2% vitamin C, and whey protein powder supplementing the remainder to 100%.

[0123] The vitamin C is L-ascorbic acid.

[0124] The preparation process of the composite peptide nutritional composition for promoting wound healing comprises the following steps:

[0125] S1. Dry-mixing fish collagen peptide, bovine spleen peptide, blood protein oligopeptide, albumin peptide, prebiotics and whey protein matrix in a mixer, crushing to a particle size of ≤40 mesh to obtain a mixture;

[0126] S2. Add ginseng powder, compound fermentation extract, and plant extract to the mixture in sequence and mix under nitrogen protection to obtain the product.

[0127] In step S1, the specific conditions for dry mixing granulation are: rotation speed of 20 rpm, mixing time of 10 min, roller pressure of 65 kN, and roller spacing of 1.0 mm.

[0128] In step S2, the specific conditions for mixing are: a rotation speed of 10 rpm and a mixing time of 15 min.

[0129] Example 2

[0130] The only difference between this embodiment and embodiment 1 is that the collagen peptide raw material is tilapia scale.

[0131] Comparative Example 1

[0132] The only difference between this comparative example and Example 1 is as follows: A2, water is added to the demineralized fish scales and the pH is adjusted to 7, alkaline protease and flavor protease are added, and after reacting at 25°C for 4 hours, the enzymes are inactivated, and the product is ultrafiltered using an ultrafiltration membrane and then freeze-dried.

[0133] Comparative Example 2

[0134] The only difference between this comparative example and Example 1 is as follows: A2, water is added to the demineralized fish scales and the pH is adjusted to 7, alkaline protease is added, and after reacting at 25°C for 2 hours, the enzyme is inactivated, and the product is ultrafiltered using an ultrafiltration membrane and then freeze-dried.

[0135] Comparative Example 3

[0136] The only difference between this comparative example and Example 1 is that the preparation method of the composite fermentation extract comprises the following steps: inoculating yeast into culture medium 1, shaking and fermenting at 30°C and 150 rpm for 72 hours to obtain fermentation product 1, adding Centella asiatica powder to obtain culture medium 2, inoculating mold, shaking and fermenting at 28°C and 200 rpm for 96 hours to obtain fermentation product 2, inoculating Lactobacillus plantarum, and anaerobically fermenting at 37°C for 48 hours, filtering through a ceramic membrane with a molecular weight cutoff of 50 kDa, vacuum concentrating at 45°C to a solid content of 30%, and spray drying to obtain the product.

[0137] The added amount of the Centella asiatica powder is 3% of the mass of the fermentation product 1.

[0138] The molds include Monascus and Aspergillus niger.

[0139] The inoculum size of Monascus was 1.5×10 7 CFU / g medium 2; the inoculum size of Aspergillus niger was 3×10 6 CFU / g culture medium 2.

[0140] The inoculation amount of the plant lactobacillus was 5×10 6 CFU / g fermentation product 2.

[0141] Comparative Example 4

[0142] The difference between this comparative example and Example 1 is that the culture medium 1, calculated by weight percentage, includes 5% soybean meal, 2.5% sucrose, and the balance is water to 100%.

[0143] Comparative Example 5

[0144] The only difference between this comparative example and Example 1 is that the preparation method of the composite fermentation extract comprises the following steps:

[0145] (1) Yeast was inoculated into culture medium 1, and fermentation was carried out at 30°C and 150 rpm for 72 h to obtain fermentation product 1;

[0146] (2) Mixing culture medium 2 and Cordyceps mycelium to obtain culture medium 2, inoculating Cordyceps militaris and Cordyceps militaris, and culturing at 23°C for 10 days to obtain fermentation product 2;

[0147] (3) Mixing medium 3 and Centella asiatica powder to obtain medium 3, inoculating mold, and fermenting at 28°C and 200 rpm for 96 hours to obtain fermentation product 3;

[0148] (4) Fermentation product 1, fermentation product 2, and fermentation product 3 were mixed to obtain culture medium 4, which was inoculated with Lactobacillus plantarum and anaerobically fermented at 37°C for 48 h. The culture medium was then filtered through a ceramic membrane with a molecular weight cutoff of 50 kDa, vacuum concentrated at 45°C to a solid content of 30%, and spray-dried to obtain the product.

[0149] The culture medium 2 and the culture medium 3, calculated by weight percentage, both contain 5% soybean meal, 2.5% sucrose, and water to make up the balance to 100%.

[0150] The amount of the Cordyceps mycelium added is consistent with the mass of culture medium 2.

[0151] The added amount of the Centella asiatica powder is 3% of the mass of the culture medium 3.

[0152] The inoculation amount of the plant lactobacillus was 5×10 6 CFU / g culture medium 4.

[0153] Comparative Example 6

[0154] The only difference between this comparative example and Example 1 is that the mold is Monascus purpureus, and the inoculation amount is 2×10 7 CFU / g culture medium 3.

[0155] Comparative Example 7

[0156] The only difference between this comparative example and Example 1 is that the green tea extract is prepared by the following steps: drying green tea, grinding the green tea and passing it through a 60-mesh sieve to obtain green tea powder, adding 70% ethanol aqueous solution by mass, performing ultrasonic-assisted extraction and filtering, concentrating by rotary evaporation at 45°C to twice the mass of the green tea powder, freeze-drying, and pulverizing and passing it through a 100-mesh sieve to obtain the green tea extract.

[0157] Experimental design

[0158] Animal Grouping: Male BALB / c mice (6-8 weeks old, 20 ± 2 g) were randomly divided into 10 groups, with 20 mice in each group. Experimental Groups 1-9 were administered with the composite peptide nutritional compositions prepared in Examples 1-2 and Comparative Examples 1-7 at a dose of 1 g / kg / day, dissolved in water and administered orally. Experimental Group 10 was administered as a blank group, with an equal amount of normal saline administered orally.

[0159] Modeling and drug administration: After shaving the back, a full-thickness skin defect was created with an 8 mm punch. Immediately after surgery, oral administration was started once a day for 14 consecutive days.

[0160] Observation indicators: ① Wound healing rate: The wound area was measured daily, and the healing rate was calculated as (initial area - residual area) / initial area × 100%; ② Histological analysis: Wound tissue was obtained on the 7th day, and the percentage of collagen deposition area was calculated by Masson trichrome staining; ③ Inflammatory factor detection: Blood was collected from the orbital venous plexus on the 7th day, and the blood was allowed to stand at room temperature for 30 minutes, then centrifuged to separate the serum, and the TNF-α concentration in the serum was determined by ELISA.

[0161] The evaluation criteria are shown in Table 1.

[0162] Table 1 Evaluation criteria

[0163] The evaluation results are shown in Table 2.

[0164] Table 2 Measurement results

[0165] According to the above data, the composite peptide nutritional composition prepared in Examples 1-2 of the present invention has a fast wound healing speed and significant effect, a high proportion of collagen deposition, and significantly reduces the level of TNF-α. Among them, the hydroxyproline content in tilapia scale collagen peptide is lower than that in cod skin, which has a certain effect on the wound healing effect, but it can be ignored, and the overall total effective rate can still reach 100%. In comparative example 1, mixed enzymatic hydrolysis leads to substrate competition, destroys the molecular weight distribution of collagen peptides, and reduces the healing effect; in comparative example 2, the lack of flavor protease leads to the unhydrolyzed collagen helical region, which reduces the proportion of peptides below 3000Da; in comparative example 3, the cordyceps stage is missing, and cordycepin is not produced; in comparative example 4, selenomethionine is missing in the yeast fermentation product, resulting in a weakened anti-inflammatory effect and affecting the overall healing effect; in comparative example 5, the fermentation products of the three stages are mixed and inoculated with plant lactobacillus, which destroys the metabolic cascade, leads to competitive inhibition of the bacterial flora, and reduces the content of effective products, affecting the healing effect; in comparative example 6, after the lack of Aspergillus niger, the conversion rate of asiaticoside is insufficient, the content of triterpenoid saponins is reduced, the anti-inflammatory effect is reduced, and the healing effect is affected; in comparative example 7, the green tea extract is not embedded, resulting in its effective ingredients such as EGCG being degraded by gastric acid, reducing the antioxidant and anti-inflammatory effects, and affecting the wound healing efficiency. Therefore, the composite peptide nutritional composition prepared using the raw materials and methods described in this application can promote wound healing. The composite peptide nutritional composition heals quickly, achieving significant healing on the 7th day and almost complete healing on the 14th day. At the same time, the collagen deposition area is large and the TNF-α concentration decreases rapidly, indicating that by increasing the content of pro-healing peptide segments, increasing the key active ingredients with anti-inflammatory and repair effects, and increasing the bioavailability of effective ingredients, it can synergistically promote the inflammatory, proliferative, and remodeling phases of wound healing, improve the speed and quality of wound healing, and provide an efficient solution for postoperative wounds, trauma, ulcers, dermatitis and other wounds.

[0166] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A composite peptide nutritional composition for promoting wound healing, characterized in that: The raw materials for its preparation, calculated by weight percentage, include 15%-25% fish collagen peptide, 0.03%-0.05% bovine spleen peptide, 0.10%-0.30% blood protein oligopeptide, 0.10%-0.30% albumin peptide, 0.01%-0.03% ginseng powder, 15%-20% compound fermentation extract, 10%-20% plant extract, 20%-25% prebiotics, and whey protein matrix supplemented with the remainder to 100%; The preparation method of the fish collagen peptide comprises the following steps: A1. Mix the collagen peptide raw material with sodium citrate solution, stir, centrifuge, and then wash with water to obtain demineralized fish scales; A2. Add water to the demineralized fish scales and adjust the pH, add alkaline protease, react, add flavor protease to react, inactivate the enzyme, ultrafilter using an ultrafiltration membrane, and then freeze-dry.

2. The composite peptide nutritional composition for promoting wound healing according to claim 1, characterized in that: The addition amount of the alkaline protease is 300-400 U / g collagen peptide raw material; the addition amount of the flavor protease is 300-400 U / g collagen peptide raw material.

3. The composite peptide nutritional composition for promoting wound healing according to claim 1, characterized in that: The preparation method of the composite fermentation extract comprises the following steps: inoculating yeast on a culture medium 1, obtaining fermentation product 1 after fermentation, adding cordyceps mycelium to obtain a culture medium 2, inoculating cicadae and cordyceps militaris, obtaining fermentation product 2 after culture, adding Centella asiatica powder to obtain a culture medium 3, inoculating mold, fermenting to obtain fermentation product 3, inoculating Lactobacillus plantarum, after fermentation, membrane filtration, concentration, and spray drying to obtain the product.

4. The composite peptide nutritional composition for promoting wound healing according to claim 3, characterized in that: The yeast includes Candida utilis.

5. The composite peptide nutritional composition for promoting wound healing according to claim 3, characterized in that: The culture medium 1 comprises, by weight percentage, 5% soybean meal, 3%-4% sucrose, 0.01% sodium selenite, and water to make up the balance to 100%.

6. The composite peptide nutritional composition for promoting wound healing according to claim 1, characterized in that: The plant extracts include green tea extract and rhodiola rosea extract.

7. The composite peptide nutritional composition for promoting wound healing according to claim 1, characterized in that: The prebiotics include resistant dextrin and galacto-oligosaccharide.

8. The composite peptide nutritional composition for promoting wound healing according to claim 1, characterized in that: The whey protein matrix comprises, by weight percentage of the whey protein matrix, 5%-10% lactose, 1%-2% vitamin C, and whey protein powder as the remainder to 100%.

9. A process for preparing the composite peptide nutritional composition for promoting wound healing according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1, granulating and crushing fish collagen peptide, bovine spleen peptide, blood protein oligopeptide, albumin peptide, prebiotics and whey protein matrix to obtain a mixture; S2. Add ginseng powder, compound fermentation extract, and plant extract to the mixture in sequence, and mix them to obtain the product.

10. Use of the complex peptide nutritional composition for promoting wound healing according to any one of claims 1 to 8 in the field of medical nutritional food.

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