Preparation method of cubilose peptide for promoting cell repair

Abstract

The invention relates to the technical field of bioactive peptides, and particularly discloses a preparation method of a cubilose peptide for promoting cell repair, which comprises the following steps: crushing cleaned and pretreated cubilose, mixing the crushed cubilose with deionized water, and grinding for 20-30 minutes to obtain a cubilose crude extract; adding mannitol and ascorbic acid into the cubilose crude extract, heating to 70-80 DEG C, keeping the temperature for 2-3 hours, and performing enzymolysis reaction in the presence of papain; after enzymolysis, carrying out high-temperature enzyme deactivation, cooling to room temperature to obtain zymolyte, adding glycerol, carrying out ultrasonic treatment, and filtering to obtain cubilose peptide crude liquid; centrifuging the cubilose peptide crude liquid, and freeze-drying to obtain cubilose peptide; according to the preparation method, the extraction rate of effective active ingredients is high, the complexity of the preparation process is reduced, and the stability of the cubilose peptide product quality is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of bioactive peptides, and more specifically, it relates to a method for preparing bird's nest peptides that promote cell repair. Background Art

[0002] As a traditional nourishing food material, bird's nest has the reputation of "sacred nourishing product" in our country. In recent years, with the development of biotechnology and nutrition, bird's nest has been further processed and extracted, and its active ingredients have been efficiently utilized, forming various functional products, among which bird's nest peptides are one of them. Bird's nest peptides have attracted wide attention due to their unique biological activities and health care functions.

[0003] The primary step in preparing bird's nest peptides is to select high-quality bird's nest as the raw material. High-quality bird's nest is generally harvested from the natural nests of swiftlets and undergoes strict screening and testing to ensure its nutritional value and safety. Subsequently, the bird's nest is pretreated, including steps such as washing and removing impurities, to prepare for the subsequent extraction process.

[0004] Patent CN115896219B relates to a method for preparing bird's nest peptides with the effects of promoting cell repair, high moisturizing and whitening, and the preparation method includes: mixing bird's nest and water, stewing to prepare cooked bird's nest; homogenizing the cooked bird's nest to prepare bird's nest slurry; enzymatically hydrolyzing the bird's nest slurry with alkaline protease, inactivating the enzyme to prepare an enzymatic hydrolysate; centrifuging the enzymatic hydrolysate, collecting the supernatant to prepare a crude bird's nest peptide solution; ultrafiltering the crude bird's nest peptide solution, collecting the ultrafiltrate to prepare bird's nest peptides; the conditions for centrifugation include: a speed of 10000 rpm to 15000 rpm and a time of 10 min to 30 min; the membrane pore size used for ultrafiltration is 100 nm to 200 nm. The bird's nest peptides prepared by this preparation process are basically consistent with the bird's nest raw material in terms of composition, inheriting and retaining the excellent characteristics of the bird's nest raw material, and in terms of functional properties, they have the functions of cell antioxidant, cell moisturizing, cell scratch repair, and proliferation.

[0005] Although the aforementioned bird's nest peptide preparation scheme has its uniqueness and potential application value, in practical applications, attention still needs to be paid to the complexity of its preparation process and the stability of product quality. In view of these problems, the present invention provides a method for preparing bird's nest peptides that promote cell repair, further studies and optimizes the preparation process, and improves product quality and performance. Summary of the Invention

[0006] In order to solve the problem of low extraction rate of effective active ingredients in the existing extraction method, and at the same time reduce the complexity of the preparation process and improve the stability of the quality of bird's nest peptide products, the present invention provides a method for preparing bird's nest peptides that promote cell repair.

[0007] In a first aspect, the present invention provides a method for preparing bird's nest peptide for promoting cell repair, adopting the following technical solution:

[0008] A method for preparing bird's nest peptide for promoting cell repair, comprising the following steps:

[0009] (1) After cleaning and pre-treating the bird's nest and then pulverizing it, take the pulverized bird's nest and deionized water, mix and grind them for 20 - 30 min to obtain a crude bird's nest extract;

[0010] (2) Add mannitol and ascorbic acid to the crude bird's nest extract, then raise the temperature to 70 - 80 °C and keep it warm for 2 - 3 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0011] (3) After enzymatic hydrolysis, inactivate the enzyme at high temperature and then cool it to room temperature to obtain an enzymatic hydrolysate, add glycerol and then carry out ultrasonic treatment, and filter to obtain a crude bird's nest peptide solution;

[0012] (4) Centrifuge the crude bird's nest peptide solution, and then obtain the bird's nest peptide after freeze-drying.

[0013] Further, the pre-treatment step includes: rinsing the surface of the bird's nest with clean water at room temperature, then soaking the bird's nest in clean water to ensure softening of the bird's nest. During this period, the soaking water can be changed 2 - 3 times. After picking out impurities, dry the water of the bird's nest with a gauze and air-dry it.

[0014] Further, the weight ratio of the bird's nest, deionized water, mannitol and ascorbic acid is 1:(3 - 4):(4 - 6):(1 - 2).

[0015] Further, in the step (2), the addition amount of papain relative to the pulverized bird's nest is 6 - 8 U / g; the enzymatic hydrolysis temperature is 45 - 55 °C, the pH value is 6.5 - 7.5, and the enzymatic hydrolysis time is 6 - 8 h.

[0016] Further, in the step (3), the high-temperature enzyme inactivation step is: raise the temperature to 85 - 95 °C and keep it for 20 - 30 min.

[0017] Further, in the step (3), the ultrasonic power is 500 - 600 W, the ultrasonic frequency is 24 - 26 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1 - 2 h.

[0018] Further, in the step (3), the weight ratio of the enzymatic hydrolysate and glycerol is 1:(4 - 5).

[0019] Further, in the step (4), the centrifugation speed is 1400 - 1600 r / min, and the centrifugation time is 10 - 20 min.

[0020] Further, the lyophilization conditions in step (4) are as follows: the freezing temperature is -15 ± 5°C, the vacuum degree is 10 - 12 Pa, and the time is 20 - 22 h.

[0021] In a second aspect, the present invention provides a bird's nest peptide, adopting the following technical solution:

[0022] A bird's nest peptide is obtained by extracting with the method for preparing a bird's nest peptide for promoting cell repair as described above.

[0023] In a third aspect, the present invention provides an application of a bird's nest peptide, adopting the following technical solution:

[0024] An application of the bird's nest peptide as described above in the preparation of skin care products, functional foods or drugs.

[0025] In summary, the present invention has the following beneficial effects:

[0026] In the present invention, the bird's nest is pre-treated by washing, crushed and then ground in deionized water, effectively breaking the binding force between macromolecules such as proteins in the bird's nest, decomposing these macromolecular substances into smaller particles or fragments; this also helps to release the active ingredients inside the bird's nest, making it easier to be dissolved and extracted in the subsequent extraction process; and the ground bird's nest particles are smaller, with a larger surface area, increasing the contact area with the solvent, thereby improving the solubility of the active ingredients in the solvent and the extraction efficiency;

[0027] In the present invention, mannitol and ascorbic acid are added to the crude bird's nest extract. These two components not only enhance the stability of peptide molecules but also play multiple protective roles. During the enzymatic hydrolysis process, mannitol and ascorbic acid can prevent the active ingredients in the bird's nest from being destroyed and at the same time promote the enzymatic hydrolysis reaction, thereby increasing the extraction rate of effective active ingredients;

[0028] In the present invention, the enzymatic hydrolysate is mixed with glycerol and ultrasonicated. Glycerol plays multiple roles in this process: first, using its enzyme-inactivating effect, it can quickly inactivate the enzyme in the enzymatic hydrolysis reaction, ensuring that the enzymatic hydrolysis reaction stops completely; second, it further breaks the protein structure in the bird's nest, making it easier to be enzymatically hydrolyzed into small peptides; third, as a solvent and stabilizer, it further extracts the active ingredients in the bird's nest and, through interactions such as forming hydrogen bonds, binds tightly to the active ingredients to prevent them from being oxidized or degraded and maintain their stability;

[0029] The present invention uses papain for enzymatic hydrolysis. By precisely controlling the addition amount of papain, the enzymatic hydrolysis temperature, pH value and enzymatic hydrolysis time, it can more effectively decompose the proteins in the bird's nest into small peptides, thereby releasing more effective active ingredients; and the enzymatic hydrolysis process is mild and efficient, which can retain the nutritional components and biological activities in the bird's nest to the greatest extent and improve the extraction rate of the bird's nest peptide;

[0030] The preparation method of the present invention has been comprehensively optimized, including steps such as pretreatment, enzymatic hydrolysis, enzyme inactivation, ultrasonic treatment, filtration, and centrifugation. These steps are carefully designed and precisely controlled, making the entire preparation process more concise, reducing unnecessary operating steps and complexity. This optimization not only reduces the complexity of the preparation process but also improves the preparation efficiency and product quality. Specific Embodiments

[0031] The present invention will be further described in detail below in conjunction with embodiments, but not limited thereto. According to the following detailed description and embodiments, other features and advantages of the present invention will become apparent, and the detailed description and embodiments should not be construed as restrictive.

[0032] Embodiment 1

[0033] A method for preparing bird's nest peptide for promoting cell repair includes the following steps:

[0034] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 3 times. After picking out impurities, dry the water of the bird's nest with a gauze and air-dry it;

[0035] (2) After cleaning and pretreating the bird's nest, crush it, and then mix the crushed bird's nest with deionized water and grind for 25 min to obtain a crude bird's nest extract;

[0036] (3) Add mannitol and ascorbic acid to the crude bird's nest extract, heat to 75 °C, and keep warm for 2.5 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0037] The weight ratio of the bird's nest, deionized water, mannitol, and ascorbic acid is 1:3.5:5:1.5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 h;

[0038] (4) After enzymatic hydrolysis, raise the temperature to 90 °C and keep it for 25 min for high-temperature enzyme inactivation, then cool to room temperature to obtain an enzymolysate. Add glycerol and carry out ultrasonic treatment, and then filter to obtain a crude bird's nest peptide solution; the weight ratio of the enzymolysate to glycerol is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 h;

[0039] (5) Centrifuge the crude bird's nest peptide solution, and then obtain the bird's nest peptide after freeze-drying; the centrifugation speed is 1500 r / min, and the centrifugation time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0040] Embodiment 2

[0041] A preparation method of bird's nest peptide for promoting cell repair, comprising the following steps:

[0042] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 2 times. After removing impurities, blot the water from the bird's nest with a gauze and air-dry it;

[0043] (2) After crushing the bird's nest that has undergone cleaning pretreatment, take the crushed bird's nest and deionized water and mix and grind them for 20 min to obtain a crude bird's nest extract;

[0044] (3) Add mannitol and ascorbic acid to the crude bird's nest extract, then raise the temperature to 80 °C and keep it warm for 2 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0045] The weight ratio of the bird's nest, deionized water, mannitol and ascorbic acid is 1:4:4:2; the addition amount of papain relative to the crushed bird's nest is 6 U / g; the enzymatic hydrolysis temperature is 55 °C, the pH value is 6.5, and the enzymatic hydrolysis time is 8 h;

[0046] (4) After enzymatic hydrolysis, raise the temperature to 85 °C and keep it for 30 min for high-temperature enzyme inactivation, then cool it to room temperature to obtain an enzymolysate. Add glycerol and carry out ultrasonic treatment, and then filter to obtain a crude bird's nest peptide solution; the weight ratio of the enzymolysate and glycerol is 1:4; the ultrasonic power is 600 W, the ultrasonic frequency is 24 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 2 h;

[0047] (5) Centrifuge the crude bird's nest peptide solution, and then obtain the bird's nest peptide after freeze-drying; the centrifugation speed is 1400 r / min, and the centrifugation time is 20 min; the freeze-drying conditions are: the freezing temperature is -10 °C, the vacuum degree is 12 Pa, and the time is 20 h.

[0048] Example 3

[0049] A preparation method of bird's nest peptide for promoting cell repair, comprising the following steps:

[0050] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 3 times. After removing impurities, blot the water from the bird's nest with a gauze and air-dry it;

[0051] (2) After crushing the bird's nest that has undergone cleaning pretreatment, take the crushed bird's nest and deionized water and mix and grind them for 30 min to obtain a crude bird's nest extract;

[0052] (3) Add mannitol and ascorbic acid to the crude bird's nest extract, then raise the temperature to 70 °C and keep it warm for 3 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0053] The weight ratio of bird's nest, deionized water, mannitol and ascorbic acid is 1:3:6:1; the addition amount of papain relative to the crushed bird's nest is 8 U / g; the enzymatic hydrolysis temperature is 45 °C, the pH value is 7.5, and the enzymatic hydrolysis time is 6 h;

[0054] (4) After enzymatic hydrolysis, raise the temperature to 95 °C and keep it for 20 min to inactivate the enzyme at high temperature, then cool it to room temperature to obtain the enzymatic hydrolysate. Add glycerol and carry out ultrasonic treatment, and then filter to obtain the crude bird's nest peptide solution; the weight ratio of the enzymatic hydrolysate to glycerol is 1:5; the ultrasonic power is 500 W, the ultrasonic frequency is 26 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1 h;

[0055] (5) Centrifuge the crude bird's nest peptide solution, and then freeze-dry it to obtain bird's nest peptide; the centrifugation speed is 1600 r / min and the centrifugation time is 10 min; the freeze-drying conditions are: the freezing temperature is -20 °C, the vacuum degree is 10 Pa, and the time is 22 h.

[0056] Example 4

[0057] A method for preparing bird's nest peptide for promoting cell repair, comprising the following steps:

[0058] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure that the bird's nest softens. During this period, the soaking water can be changed 2 times. After picking out the impurities, dry the bird's nest with a gauze and air-dry it;

[0059] (2) After cleaning and pretreating the bird's nest, crush it, and then mix the crushed bird's nest and deionized water and grind for 20 min to obtain the crude bird's nest extract;

[0060] (3) Add mannitol and ascorbic acid to the crude bird's nest extract, then raise the temperature to 70 °C and keep it warm for 2 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0061] The weight ratio of bird's nest, deionized water, mannitol and ascorbic acid is 1:3:4:1; the addition amount of papain relative to the crushed bird's nest is 6 U / g; the enzymatic hydrolysis temperature is 45 °C, the pH value is 6.5, and the enzymatic hydrolysis time is 6 h;

[0062] (4) After enzymatic hydrolysis, raise the temperature to 85 °C and keep it for 20 min to inactivate the enzyme at high temperature, then cool it to room temperature to obtain the enzymatic hydrolysate. Add glycerol and carry out ultrasonic treatment, and then filter to obtain the crude bird's nest peptide solution; the weight ratio of the enzymatic hydrolysate to glycerol is 1:4; the ultrasonic power is 500 W, the ultrasonic frequency is 24 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1 h;

[0063] (5) Centrifuge the crude bird's nest peptide solution, and then freeze-dry it to obtain the bird's nest peptide; the centrifuge speed is 1400 r / min, and the centrifuge time is 10 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 10 Pa, and the time is 20 h.

[0064] Comparative Example 1

[0065] A method for preparing bird's nest peptide for promoting cell repair, comprising the following steps:

[0066] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening of the bird's nest. During this period, the soaking water can be changed 3 times. After removing impurities, dry the water on the bird's nest with a gauze and air-dry it;

[0067] (2) After crushing the bird's nest that has been cleaned and pretreated, take the crushed bird's nest and deionized water and mix them to obtain a crude bird's nest extract;

[0068] (3) Add mannitol and ascorbic acid to the crude bird's nest extract, then raise the temperature to 75 °C and keep it warm for 2.5 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0069] The weight ratio of the bird's nest, deionized water, mannitol and ascorbic acid is 1:3.5:5:1.5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 h;

[0070] (4) After enzymatic hydrolysis, raise the temperature to 90 °C and keep it for 25 min to inactivate the enzyme at high temperature, then cool it to room temperature to obtain an enzymatic hydrolysate. Add glycerol and then carry out ultrasonic treatment, and filter to obtain a crude bird's nest peptide solution; the weight ratio of the enzymatic hydrolysate and glycerol is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 h;

[0071] (5) Centrifuge the crude bird's nest peptide solution, and then freeze-dry it to obtain the bird's nest peptide; the centrifuge speed is 1500 r / min, and the centrifuge time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0072] Comparative Example 2

[0073] A method for preparing bird's nest peptide for promoting cell repair, comprising the following steps:

[0074] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening of the bird's nest. During this period, the soaking water can be changed 3 times. After removing impurities, dry the water on the bird's nest with a gauze and air-dry it;

[0075] (2) After crushing the bird's nest that has undergone cleaning and pretreatment, take the crushed bird's nest and deionized water, mix them and grind for 25 minutes to obtain a crude extract of the bird's nest;

[0076] (3) Add ascorbic acid to the crude extract of the bird's nest, then raise the temperature to 75 °C and keep it warm for 2.5 hours, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0077] The weight ratio of the bird's nest, deionized water, and ascorbic acid is 1:3.5:1.5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 hours;

[0078] (4) After enzymatic hydrolysis, raise the temperature to 90 °C and keep it for 25 minutes to inactivate the enzyme at high temperature, then cool it to room temperature to obtain an enzymatic hydrolysate. Add glycerol and carry out ultrasonic treatment, and then filter to obtain a crude solution of bird's nest peptides; the weight ratio of the enzymatic hydrolysate to glycerol is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 hours;

[0079] (5) Centrifuge the crude solution of bird's nest peptides, and then freeze-dry to obtain bird's nest peptides; the centrifugation speed is 1500 r / min, and the centrifugation time is 15 minutes; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 hours.

[0080] Comparative Example 3

[0081] A method for preparing bird's nest peptides for promoting cell repair, comprising the following steps:

[0082] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening of the bird's nest. During this period, the soaking water can be changed 3 times. After picking out impurities, dry the water on the bird's nest with a gauze and air-dry it;

[0083] (2) After crushing the bird's nest that has undergone cleaning and pretreatment, take the crushed bird's nest and deionized water, mix them and grind for 25 minutes to obtain a crude extract of the bird's nest;

[0084] (3) Add mannitol to the crude extract of the bird's nest, then raise the temperature to 75 °C and keep it warm for 2.5 hours, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0085] The weight ratio of the bird's nest, deionized water, and mannitol is 1:3.5:5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 hours;

[0086] (4) After enzymatic hydrolysis, the temperature is raised to 90 °C and maintained for 25 min for high-temperature inactivation of enzymes, then cooled to room temperature to obtain the enzymatic hydrolysate. Glycerol is added, followed by ultrasonic treatment, and then filtration to obtain the crude bird's nest peptide solution; the weight ratio of the enzymatic hydrolysate to glycerol is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 h.

[0087] (5) The crude bird's nest peptide solution is centrifuged and then freeze-dried to obtain the bird's nest peptide; the centrifugation speed is 1500 r / min and the centrifugation time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0088] Comparative Example 4

[0089] A method for preparing bird's nest peptide for promoting cell repair, comprising the following steps:

[0090] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 3 times. After removing impurities, blot the water from the bird's nest with a gauze and air-dry it.

[0091] (2) After the bird's nest that has been cleaned and pretreated is pulverized, the pulverized bird's nest and deionized water are mixed and ground for 25 min to obtain the crude bird's nest extract.

[0092] (3) Mannitol and ascorbic acid are added to the crude bird's nest extract, and the temperature is raised to 75 °C and kept for 2.5 h, and an enzymatic hydrolysis reaction is carried out in the presence of alkaline protease.

[0093] The weight ratio of the bird's nest, deionized water, mannitol, and ascorbic acid is 1:3.5:5:1.5; the addition amount of alkaline protease relative to the pulverized bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 h.

[0094] (4) After enzymatic hydrolysis, the temperature is raised to 90 °C and maintained for 25 min for high-temperature inactivation of enzymes, then cooled to room temperature to obtain the enzymatic hydrolysate. Glycerol is added, followed by ultrasonic treatment, and then filtration to obtain the crude bird's nest peptide solution; the weight ratio of the enzymatic hydrolysate to glycerol is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 h.

[0095] (5) The crude bird's nest peptide solution is centrifuged and then freeze-dried to obtain the bird's nest peptide; the centrifugation speed is 1500 r / min and the centrifugation time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0096] Comparative Example 5

[0097] A preparation method of bird's nest peptide for promoting cell repair, comprising the following steps:

[0098] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 3 times. After picking out impurities, dry the water of the bird's nest with a gauze and air-dry it;

[0099] (2) After crushing the bird's nest that has been cleaned and pretreated, take the crushed bird's nest and deionized water and mix and grind them for 25 min to obtain a crude extract of bird's nest;

[0100] (3) Add mannitol and ascorbic acid to the crude extract of bird's nest, then raise the temperature to 75 °C and keep it warm for 2.5 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0101] The weight ratio of the bird's nest, deionized water, mannitol and ascorbic acid is 1:3.5:5:1.5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymatic hydrolysis temperature is 50 °C, the pH value is 7, and the enzymatic hydrolysis time is 7 h;

[0102] (4) After enzymatic hydrolysis, raise the temperature to 90 °C and keep it for 25 min for high-temperature enzyme inactivation, then cool it to room temperature to obtain an enzymatic hydrolysate. Add deionized water and carry out ultrasonic treatment, and then filter to obtain a crude solution of bird's nest peptide; the weight ratio of the enzymatic hydrolysate and deionized water is 1:4.5; the ultrasonic power is 550 W, the ultrasonic frequency is 25 kHz, the ultrasonic temperature is at room temperature, and the ultrasonic time is 1.5 h;

[0103] (5) Centrifuge the crude solution of bird's nest peptide, and then obtain the bird's nest peptide after freeze-drying; the centrifugation speed is 1500 r / min and the centrifugation time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0104] Comparative Example 6

[0105] A preparation method of bird's nest peptide for promoting cell repair, comprising the following steps:

[0106] (1) Pretreatment: Rinse the surface of the bird's nest with clean water at room temperature, then soak the bird's nest in clean water to ensure softening. During this period, the soaking water can be changed 3 times. After picking out impurities, dry the water of the bird's nest with a gauze and air-dry it;

[0107] (2) After crushing the bird's nest that has been cleaned and pretreated, take the crushed bird's nest and deionized water and mix and grind them for 25 min to obtain a crude extract of bird's nest;

[0108] (3) Add mannitol and ascorbic acid to the crude extract of bird's nest, then raise the temperature to 75 °C and keep it warm for 2.5 h, and carry out an enzymatic hydrolysis reaction in the presence of papain;

[0109] The weight ratio of bird's nest, deionized water, mannitol and ascorbic acid is 1:3.5:5:1.5; the addition amount of papain relative to the crushed bird's nest is 7 U / g; the enzymolysis temperature is 50 °C, the pH value is 7, and the enzymolysis time is 7 h;

[0110] (4) After enzymolysis, the temperature is raised to 90 °C and maintained for 25 min for high-temperature inactivation of the enzyme, then cooled to room temperature to obtain the enzymolysis product. Glycerol is added and mixed, and stirred at a rotation speed of 200 r / min for 1.5 h, and then filtered to obtain the crude bird's nest peptide solution; the weight ratio of the enzymolysis product to glycerol is 1:4.5;

[0111] (5) The crude bird's nest peptide solution is centrifuged and then freeze-dried to obtain bird's nest peptide; the centrifugation speed is 1500 r / min and the centrifugation time is 15 min; the freeze-drying conditions are: the freezing temperature is -15 °C, the vacuum degree is 11 Pa, and the time is 21 h.

[0112] Test example: The bird's nest peptides prepared in Examples 1-4 and Comparative Examples 1-6 of the present invention are respectively dissolved in a solvent to obtain uniformly mixed test samples 1-4 and comparative test samples 1-6. Specifically, 1 g of bird's nest peptide is dissolved in 5 g of the solvent, wherein the solvent is composed of 80 wt% deionized water and 20 wt% sodium hyaluronate.

[0113] Performance test:

[0114] I. Evaluation of the effect of cell repair

[0115] Test subjects: In the same region and at the same time, 140 female volunteers with skin aging were screened, aged 38-52 years old.

[0116] Inclusion criteria: Mild or moderate wrinkles, dull skin, decreased elasticity of collagen fibers, dark brown spots, etc. appear around the forehead. And the subjects are in good health, have no history of skin diseases and allergies, are not sensitive skin, and can use skin care products according to the specifications.

[0117] Usage method: The 140 volunteers are randomly divided into 7 groups, with 20 people in each group. The specific usage method is as follows: After cleaning the facial skin in the morning and evening every day, dry it, and evenly apply 3 g of the test sample on the facial skin with hands, and use it continuously for one month.

[0118] Evaluation method: Using a VISIA tester, the skin conditions of the subjects were tested before and after using for 1 month, and the wrinkle reduction rate, skin tone brightening rate and skin elasticity improvement rate of each group of subjects were calculated. The test results are shown in Table 1.

[0119] Table 1:

[0120] Wrinkle reduction rate (%) Skin tone brightening rate (%) Skin elasticity improvement rate (%) Example 1 36.5 33.4 35.7 Example 2 36.3 32.5 35.4 Example 3 35.8 32.1 34.8 Example 4 36.1 32.9 35.1 Comparative Example 1 20.5 20.6 20.1 Comparative Example 2 16.8 18.2 16.2 Comparative Example 3 15.2 16.7 14.3 Comparative Example 4 21.4 22.4 18.7 Comparative Example 5 12.1 11.4 10.6 Comparative Example 6 13.5 13.0 12.8

[0121] II. Stability Evaluation

[0122] On the basis of evaluating the effect of cell repair, the test samples were stopped being used. After one month, the VISIA tester was used again to measure the skin conditions of the subjects before and one month after the use, and the wrinkle reduction rate, skin tone brightening rate, and skin elasticity improvement rate of each group of subjects were calculated. The test results are shown in Table 2.

[0123] Table 2:

[0124]

[0125]

[0126] This specific embodiment is only an interpretation of the present invention and not a limitation thereof. After reading this specification, those skilled in the art may make modifications to this embodiment without creative contributions as needed, but as long as it is within the scope of the claims of the present invention, it is protected by the patent law.

Claims

1. A preparation method of bird's nest peptide for promoting cell repair, characterized in that, It includes the following steps: (1) After the cleaned and pretreated bird's nest is crushed, the crushed bird's nest and deionized water are mixed and ground for 20 - 30 min to obtain a crude extract of bird's nest; (2) Mannitol and ascorbic acid are added to the crude extract of bird's nest, and then the temperature is raised to 70 - 80 °C and kept warm for 2 - 3 h, and an enzymatic hydrolysis reaction is carried out in the presence of papain; (3) After enzymatic hydrolysis, the enzyme is inactivated at high temperature and then cooled to room temperature to obtain an enzymatic hydrolysate. Glycerol is added and then ultrasonic treatment is carried out, and then filtered to obtain a crude solution of bird's nest peptide; (4) The crude solution of bird's nest peptide is centrifuged and then freeze - dried to obtain bird's nest peptide.

2. The method for preparing bird's nest peptide for promoting cell repair according to claim 1, characterized in that, The pretreatment step includes: rinsing the surface of the bird's nest with clean water at room temperature, then soaking the bird's nest in clean water to ensure softening of the bird's nest. During this period, the soaking water can be changed 2 - 3 times. After picking out impurities, the bird's nest is blotted dry with a gauze and air - dried.

3. The method for preparing the bird's nest peptide for promoting cell repair according to claim 1, wherein The weight - part ratio of the bird's nest, deionized water, mannitol and ascorbic acid is 1:(3 - 4):(4 - 6):(1 - 2).

4. The method for preparing bird's nest peptide for promoting cell repair according to claim 1, characterized in that In the step (2), the addition amount of papain relative to the crushed bird's nest is 6 - 8 U / g; the enzymatic hydrolysis temperature is 45 - 55 °C, the pH value is 6.5 - 7.5, and the enzymatic hydrolysis time is 6 - 8 h.

5. The preparation method of the edible bird's nest peptide for promoting cell repair according to claim 1, wherein, In the step (3), the high - temperature enzyme inactivation step is: raising the temperature to 85 - 95 °C and keeping it for 20 - 30 min; in the step (3), the ultrasonic power is 500 - 600 W, the ultrasonic frequency is 24 - 26 kHz, the ultrasonic temperature is at room - temperature condition, and the ultrasonic time is 1 - 2 h.

6. The method for preparing bird's nest peptide for promoting cell repair according to claim 1, wherein, In the step (3), the weight - part ratio of the enzymatic hydrolysate and glycerol is 1:(4 - 5).

7. The method for preparing the edible bird's nest peptide for promoting cell repair according to claim 1, characterized in that In the step (4), the centrifugation speed is 1400 - 1600 r / min, and the centrifugation time is 10 - 20 min.

8. The method for preparing the edible bird's nest peptide for promoting cell repair according to claim 1, wherein, In the step (4), the freeze - drying conditions are: the freezing temperature is - 15 ± 5 °C, the vacuum degree is 10 - 12 Pa, and the time is 20 - 22 h.

9. A bird's nest peptide, characterized in that, It is obtained by extracting with the method for preparing bird's nest peptide for promoting cell repair according to any one of claims 1 - 8.

10. Use of the method for preparing bird's nest peptide according to claim 9 in the preparation of skin care products, functional foods or pharmaceuticals.

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