Preparation method of collagen peptide
Through the enzymatic decomposition process of microbial proteases and functional agents, combined with the use of Chinese medicines, the problem of low extraction efficiency of existing collagen peptides has been solved, and the preparation of high-purity and high-active collagen peptides has been achieved to meet market demand.
Patent Information
- Application Number
- CN202510621026.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-14
- Publication Date
- 2025-08-15
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
The existing collagen peptide extraction methods are inefficient, low yield and low activity, making it difficult to meet market demand.
Use reasonable raw material combination and preparation technology, use microbial proteases and functional agents for enzymatic decomposition, and combine the use of Chinese medicines and functional agents to improve the extraction rate and purity of collagen peptides.
It significantly improves the extraction rate and yield of collagen peptides, and makes the prepared collagen peptides have high purity and high activity, and has wide application prospects.
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Abstract
Description
Technical Field
[0001] The present application relates to the field of collagen technology, and more specifically, to a method for preparing collagen peptides. Background Art
[0002] Collagen is a fibrous protein composed of three intertwined polypeptide chains forming a triple helix structure. It is the primary component maintaining the elasticity of skin and muscles, forming a tight elastic network within the skin that locks in moisture and acts like a scaffold to support the skin, making it elastic, moisturized, delicate, and smooth. Collagen hydrolyzes under the influence of acids, alkalis, heat, and enzymes, producing collagen peptides, which play a vital role in our skin, bones, muscles, and connective tissues, helping to maintain skin elasticity and firmness, maintain joint mobility, and enhance bone health.
[0003] Currently, methods for extracting collagen peptides include acid extraction, alkaline extraction, hot water extraction, neutral salt hydrolysis, and enzymatic extraction. These methods all suffer from harsh extraction conditions, resulting in low product activity, low extraction rate, and low product purity. Numerous studies have confirmed that collagen peptides have numerous applications, including delaying skin aging, preventing and treating osteoporosis, providing antioxidant benefits, promoting wound healing, and enhancing immunity. Therefore, there is an urgent need to provide a method for preparing collagen peptides that can effectively improve collagen peptide extraction efficiency, enhance collagen peptide activity, and increase collagen peptide production to meet market needs. Summary of the Invention
[0004] In order to solve the problems of low efficiency, low yield and low activity in the collagen peptide extraction technology in the related art, the present application provides a method for preparing collagen peptide.
[0005] The present application provides a method for preparing collagen peptides, which specifically comprises the following steps:
[0006] S1, fish scales are crushed into fish scale powder, placed in sodium hydroxide solution and soaked, washed with water to neutrality, then placed in citric acid solution and soaked, washed with water to neutrality, dried for standby use;
[0007] S2, adding the fish scale powder obtained above to an enzyme reaction kettle, adding water and stirring to mix, then adding microbial protease and functional agent and stirring to mix, performing enzymatic hydrolysis, heating to inactivate the enzyme, and obtaining a protease hydrolyzate;
[0008] S3. Centrifuge the protein hydrolysate to obtain the supernatant, ultrafilter the supernatant, and freeze-dry to obtain collagen peptides.
[0009] Preferably, in step S1, the mass ratio of fish scale powder, sodium hydroxide solution and citric acid solution is 1:10:6.
[0010] Preferably, in step S2, the microbial protease is prepared by the following method:
[0011] (1) activating Aspergillus oryzae and inoculating the activated Aspergillus oryzae into a seed culture medium to obtain a seed solution;
[0012] (2) inoculating the seed liquid into a fermentation medium and fermenting to obtain a fermentation liquid;
[0013] (3) Centrifuging the fermentation broth, taking the supernatant, adding polyethylene glycol 6000 and NaCl to the supernatant at low temperature while stirring, letting it stand, and then centrifuging it, taking the precipitate, and dissolving the precipitate with borate buffer to obtain a crude protease extract;
[0014] (4) The crude protease extract is placed in a dialysis bag, dialyzed with Tris-HCl buffer to remove salt to obtain a protein solution, and the protein solution is freeze-dried to obtain the microbial protease.
[0015] Preferably, in step (1), the seed culture medium is prepared by the following method: peptone, glucose, sodium chloride and water are stirred and mixed in a mass ratio of 10:3:12:1000, and the pH is 6.0.
[0016] Preferably, in step (2), the fermentation medium is prepared by the following method: traditional Chinese medicine, yeast powder, glucose, magnesium sulfate and water are stirred and mixed in a mass ratio of 6-8:5-7:5:8-10:1000, and the pH is 6.0.
[0017] Preferably, the Chinese medicine is prepared by the following method: stirring and mixing astragalus and ginkgo leaves, crushing and sieving to obtain Chinese medicine powder, stirring and mixing the Chinese medicine powder with an ethanol solution with a mass concentration of 70% to obtain a mixed solution, adding bacterial enzyme liquid and bovine serum albumin to the mixed solution, stirring and mixing, sealing and fermenting, and centrifuging to remove residue to obtain the Chinese medicine.
[0018] Preferably, the mass ratio of the astragalus root, ginkgo leaf and ethanol solution is 5:2-3:20.
[0019] Preferably, the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin is 1:0.3-0.5:0.01.
[0020] Preferably, the bacterial enzyme solution is prepared by the following method: thermophilic streptococcus, amylase, bromelain and water are stirred and mixed in a mass ratio of 1:1-3:2-4:8.
[0021] Preferably, in step S2, the functional agent is prepared by the following method:
[0022] 1) Stirring mannitol, vinyl pyrrolidone and water, adding sodium percarbonate, heating, continuing stirring and mixing, cooling to room temperature, and centrifuging to obtain a functional substance;
[0023] 2) The functional substance is dispersed in deionized water (the amount is 50 times the weight of the functional substance) to form a functional substance solution, chitosan solution is added to the functional substance solution, ultrasonic dispersion is performed, and centrifugal drying is performed to obtain a functional agent.
[0024] Preferably, in step 1), the mass ratio of mannitol, vinyl pyrrolidone, water and sodium percarbonate is 12-20:7-9:50:2.
[0025] Preferably, in step 2), the mass ratio of the functional substance solution to the chitosan solution is 1-2:2.
[0026] Preferably, the chitosan solution in step 2) is prepared by stirring and mixing chitosan and a dilute acetic acid solution with a mass concentration of 3% in a mass ratio of 1:10.
[0027] In summary, this application has the following beneficial effects:
[0028] 1. This application adopts reasonable raw material combination, raw material ratio and preparation process. By adding microbial protease and functional agent during the enzymatic hydrolysis process, it not only effectively improves the extraction rate and yield of collagen peptides, but also makes the prepared collagen peptides have high purity and high activity, and has broad application prospects.
[0029] 2. This application adds a traditional Chinese medicine to the fermentation medium for preparing microbial protease. The traditional Chinese medicine raw materials of this traditional Chinese medicine are composed of astragalus and ginkgo leaves, and are prepared by two-step extraction with ethanol solution and bacterial enzyme solution. Astragalus and ginkgo leaves are compounded. On the one hand, the traditional Chinese medicine active ingredients contained in astragalus and ginkgo leaves have antioxidant properties. After the two are compounded, they can scavenge free radicals through multiple pathways, reduce the damage of oxidation to collagen, and synergistically improve the activity of collagen peptides. On the other hand, the active ingredients in astragalus and ginkgo leaves can reduce interfacial tension, allowing the extraction solvent to fully contact collagen, thereby promoting the dissolution and release of collagen, and synergistically improving the extraction rate of collagen peptides. The bacterial enzyme solution is prepared by compounding thermophilic streptococcus, amylase, and bromelain. The thermophilic streptococcus can increase the activity of amylase and bromelain, and amylase and bromelain can promote the growth of thermophilic streptococcus. The compounding of the three can not only synergistically promote the release of active ingredients of traditional Chinese medicine, improve the extraction rate and efficacy of traditional Chinese medicine, but also provide a suitable growth environment and nutrients for the growth of Aspergillus oryzae, thereby increasing the yield and activity of microbial proteases and further improving the extraction efficiency and activity of collagen peptides. The addition of bovine serum albumin during the preparation of the traditional Chinese medicine can combine with the active ingredients of the traditional Chinese medicine to form a complex, reduce the contact of the active ingredients with the external environment, and improve the stability of the active ingredients of the traditional Chinese medicine. In the subsequent fermentation broth precipitation step, polyethylene glycol 6000 is used for graded precipitation, which can effectively remove foreign proteins and co-precipitate the target protein and the active ingredients of the traditional Chinese medicine, further reducing the loss of the active ingredients of the traditional Chinese medicine and improving the effect of the traditional Chinese medicine.
[0030] 3. The present application adds a functional agent during the collagenase hydrolysis process. The functional agent is first grafted with vinyl pyrrolidone by mannitol and then coated with chitosan solution. The addition of the functional agent can improve the extraction efficiency and biological activity of collagen peptides. Mannitol is first grafted with vinyl pyrrolidone. On the one hand, the binding force of the functional agent and collagen can be enhanced, the solubility and dispersibility of collagen can be improved, thereby improving the extraction efficiency of collagen peptides. On the other hand, the interaction with subsequent chitosan solution can be enhanced, and the stability of the coated structure can be improved. The coating of chitosan solution introduces more binding sites, increases the affinity of the functional agent and collagen, thereby further improving the extraction efficiency, extraction purity and biological activity of collagen peptides. In addition, the addition of the functional agent can also improve the activity of microbial protease, enhance the interaction of microbial protease and collagen, thereby improving the enzymatic efficiency and accelerating the extraction of collagen peptides. DETAILED DESCRIPTION
[0031] The present application is further described in detail below with reference to the embodiments.
[0032] Example 1-5 provides a method for preparing collagen peptide.
[0033] Citric acid (CAS: 77-92-9, purchased from Henan Zunting Biotechnology Co., Ltd.), Aspergillus oryzae (No.: BWCC60282, purchased from Henan Wanjia Standard Material Research and Development Center), peptone (No.: JYS, purchased from Wuhan Jiyesheng Chemical Co., Ltd.), yeast powder (CAS: 119-44-8, purchased from Hebei Chuangzhiyuan Biotechnology Co., Ltd.), astragalus (No.: 0006, purchased from Bozhou Tengwang Pharmaceutical Co., Ltd.), ginkgo biloba (brand: Wufang Jiuru, purchased from Bozhou Wufang Jiuru Pharmaceutical Co., Ltd.), Streptococcus thermophilus (No.: YS-JZ12952) used in the examples and comparative examples of the present application. , purchased from Shanghai Yushao Biotechnology Co., Ltd.), amylase (catalog number: E0254, purchased from Nanjing Dulai Biotechnology Co., Ltd.), bromelain (CAS: 9001-00-7, purchased from Anhui Youtai Bioengineering Co., Ltd.), bovine serum albumin (catalog number: 0125, purchased from Hubei Shiteng Chemical Technology Co., Ltd.), mannitol (CAS: 87-78-5, purchased from Wuhan Dongkangyuan Technology Co., Ltd.), vinylpyrrolidone (catalog number: 88-12-0, purchased from Hubei Rishengchang New Material Technology Co., Ltd.), and chitosan (CAS: 9012-76-4, purchased from Shandong Haiyihua Biotechnology Co., Ltd.
[0034] Example 1
[0035] A method for preparing collagen peptides comprises the following steps:
[0036] S1, by the mass ratio of fish scale powder, sodium hydroxide solution, and citric acid solution of 1:10:6, the fish scales are crushed into fish scale powder and passed through an 80-mesh sieve, placed in a sodium hydroxide solution of 5% by mass concentration and soaked for 1 h, washed with water to neutrality, and then placed in a citric acid solution of 5% by mass concentration and soaked for 2 h after drying at 40 ° C, washed with water to neutrality, and dried for standby at 40 ° C;
[0037] S2, the obtained fish scale powder was added to the enzyme reactor, water was added and stirred at 400r / min for 30min, and then microbial protease and functional agent were added and stirred at 200r / min for 20min to carry out enzymatic hydrolysis at 40°C for 4h, and then the temperature was raised to 80°C to inactivate the enzyme for 15min to obtain a protease hydrolyzate;
[0038] S3. Centrifuge the protein hydrolysate at 8000 r / min for 10 min to obtain the supernatant, ultrafilter the supernatant using a 3 kDa ultrafiltration membrane, control the pressure at 0.4 MPa, and freeze-dry at -30°C for 8 h to obtain collagen peptides.
[0039] In step S2, the microbial protease is prepared by the following method:
[0040] (1) Activating Aspergillus oryzae was inoculated into a seed culture medium (prepared by mixing peptone, glucose, sodium chloride, and water in a mass ratio of 10:3:12:1000 at 400 rpm for 10 min, with a pH of 6.0), and culturing at 30°C for 8 h to obtain a seed solution;
[0041] (2) inoculating the seed liquid at an inoculum volume ratio of 3% into a fermentation medium (prepared by mixing Chinese medicinal materials, yeast powder, glucose, magnesium sulfate, and water in a mass ratio of 6:5:5:8:1000 at 400 rpm for 20 min, with a pH of 6.0) at a fermentation temperature of 28° C. and a rotation speed of 200 rpm for 10 h to obtain a fermentation liquid;
[0042] (3) The fermentation broth was centrifuged at 10,000 rpm for 10 min at 4°C, and the supernatant was collected. 10% polyethylene glycol 6000 was first added to the supernatant at 4°C while stirring, and the mixture was allowed to stand at 4°C for 3 h. 20% polyethylene glycol 6000 and 0.3 mol / L NaCl were then added, and the mixture was allowed to stand at 4°C for 8 h. The mixture was then centrifuged at 12,000 rpm for 30 min, and the precipitate was collected and dissolved in a borate buffer solution with a pH of 9.0 to obtain a crude protease extract.
[0043] (4) The crude protease extract was placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa, and dialyzed with a pH 7.5, 20 mmol / L Tris-HCl buffer at 4°C to desalt the obtained protein solution. The protein solution was freeze-dried at -20°C for 2 h to obtain the microbial protease.
[0044] The Chinese medicine is prepared by the following method:
[0045] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 2:5:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 100-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.3:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:1:2:8, and stirred and mixed at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 40°C for 5 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0046] In step S2, the functional agent is prepared by the following method:
[0047] 1) mannitol, vinyl pyrrolidone, water, and sodium percarbonate were stirred at 500 rpm for 20 minutes in a mass ratio of 12:7:50:2, sodium percarbonate was added, the temperature was controlled at 40° C., stirring was continued at 400 rpm under nitrogen for 4 hours, the mixture was cooled to room temperature, and centrifuged at 12,000 rpm for 10 minutes to obtain a functional compound;
[0048] 2) The functional substance was dispersed in deionized water (the amount was 50 times the mass of the functional substance) to form a functional substance solution. The mass ratio of the functional substance solution to the chitosan solution was 1:2. Chitosan solution (prepared by mixing chitosan and a 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 rpm for 30 minutes) was added to the functional substance solution. The mixture was ultrasonically dispersed for 1 hour, the ultrasonic power was controlled to 400 W, the ultrasonic frequency was controlled to 20 kHz, and the mixture was centrifuged at 10,000 rpm for 30 minutes. The mixture was then dried at 45° C. for 1 hour to obtain the functional agent.
[0049] Example 2
[0050] A method for preparing collagen peptides comprises the following steps:
[0051] S1, by the mass ratio of fish scale powder, sodium hydroxide solution, citric acid solution is 1:10:6, the fish scales are crushed into fish scale powder and passed through an 80-mesh sieve, placed in a sodium hydroxide solution of 5% by mass concentration and soaked for 1h, washed with water to neutrality, placed in a citric acid solution of 5% by mass concentration and soaked for 2.2h after drying at 40°C, washed with water to neutrality, and dried for standby at 42°C;
[0052] S2, the obtained fish scale powder was added to the enzyme reactor, water was added and stirred at 400r / min for 30min, and then microbial protease and functional agent were added and stirred at 200r / min for 20min to carry out enzymatic hydrolysis at 43°C for 4.5h, and then the temperature was raised to 80°C to inactivate the enzyme for 16min to obtain a protease hydrolyzate;
[0053] S3. Centrifuge the protein hydrolysate at 8000 r / min for 10 min to obtain the supernatant, ultrafilter the supernatant using a 3.5 kDa ultrafiltration membrane with a controlled pressure of 0.45 MPa, and freeze-dry at -30°C for 8.5 h to obtain collagen peptides.
[0054] In step S2, the microbial protease is prepared by the following method:
[0055] (1) Activating Aspergillus oryzae was inoculated into a seed culture medium (prepared by mixing peptone, glucose, sodium chloride, and water in a mass ratio of 10:3:12:1000 at 400 rpm for 10 min, with a pH of 6.0), and culturing at 30°C for 9 h to obtain a seed solution;
[0056] (2) inoculating the seed liquid at an inoculum volume ratio of 3.5% into a fermentation medium (prepared by mixing Chinese medicinal materials, yeast powder, glucose, magnesium sulfate, and water in a mass ratio of 6.5:5.5:5:8.5:1000 at 400 rpm for 20 min, with a pH of 6.0) at a fermentation temperature of 28.5° C. and a rotation speed of 200 rpm for 10 h to obtain a fermentation liquid;
[0057] (3) The fermentation broth was centrifuged at 4°C, 10500 r / min for 10 min, and the supernatant was collected. 10% polyethylene glycol 6000 was first added to the supernatant at 4°C while stirring, and the mixture was allowed to stand at 4°C for 3 h. Then, 20% polyethylene glycol 6000 and 0.3 mol / L NaCl were added, and the mixture was allowed to stand at 4°C for 9 h. The mixture was then centrifuged at 12000 r / min for 30 min, and the precipitate was collected and dissolved in a borate buffer solution with a pH of 9.0 to obtain a crude protease extract.
[0058] (4) The crude protease extract was placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa, and dialyzed with a pH 7.5, 20 mmol / L Tris-HCl buffer at 4°C to desalt the obtained protein solution. The protein solution was freeze-dried at -20°C for 2 h to obtain the microbial protease.
[0059] The Chinese medicine is prepared by the following method:
[0060] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 5:2.2:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together to pass through a 105-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.35:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:1.5:2.5:8 and stirred and mixed at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 42°C for 5.5 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine.
[0061] In step S2, the functional agent is prepared by the following method:
[0062] 1) mannitol, vinyl pyrrolidone, water, and sodium percarbonate were stirred at 500 rpm for 20 minutes in a mass ratio of 14:7.5:50:2, sodium percarbonate was added, the temperature was controlled at 45° C., stirring was continued at 400 rpm for 4.5 hours under nitrogen protection, the mixture was cooled to room temperature, and centrifuged at 12,000 rpm for 10 minutes to obtain a functional compound;
[0063] 2) The functional substance was dispersed in deionized water (the amount was 50 times the mass of the functional substance) to form a functional substance solution. The mass ratio of the functional substance solution to the chitosan solution was 1.3:2. Chitosan solution (prepared by mixing chitosan and a 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 rpm for 30 minutes) was added to the functional substance solution. Ultrasonic dispersion was performed for 1 hour. The ultrasonic power was controlled to 450 W and the ultrasonic frequency was controlled to 20 kHz. The mixture was centrifuged at 10,000 rpm for 30 minutes and then dried at 48°C for 1.2 hours to obtain the functional agent.
[0064] Example 3
[0065] A method for preparing collagen peptides comprises the following steps:
[0066] S1, by the mass ratio of fish scale powder, sodium hydroxide solution, citric acid solution is 1:10:6, the fish scales are crushed into fish scale powder and passed through an 80-mesh sieve, placed in a sodium hydroxide solution of 5% by mass concentration and soaked for 1h, washed with water to neutrality, placed in a citric acid solution of 5% by mass concentration and soaked for 2.5h after drying at 40°C, washed with water to neutrality, and dried for standby at 45°C;
[0067] S2, the obtained fish scale powder was added to the enzyme reactor, water was added and stirred at 400r / min for 30min, and then microbial protease and functional agent were added and stirred at 200r / min for 20min to carry out enzymatic hydrolysis at a temperature of 45°C for 5h, and the temperature was raised to 80°C for 17min to inactivate the enzyme to obtain a protease hydrolyzate;
[0068] S3. Centrifuge the protein hydrolysate at 8000 r / min for 10 min to obtain the supernatant, ultrafilter the supernatant using a 4 kDa ultrafiltration membrane with a pressure controlled at 0.5 MPa, and freeze-dry at -30°C for 9 h to obtain collagen peptides.
[0069] In step S2, the microbial protease is prepared by the following method:
[0070] (1) Activating Aspergillus oryzae was inoculated into a seed culture medium (prepared by mixing peptone, glucose, sodium chloride, and water in a mass ratio of 10:3:12:1000 at 400 rpm for 10 min, with a pH of 6.0), and culturing at 30° C. for 10 h to obtain a seed solution;
[0071] (2) inoculating the seed liquid at a volume ratio of 4% into a fermentation medium (prepared by mixing Chinese medicinal materials, yeast powder, glucose, magnesium sulfate, and water in a mass ratio of 7:6:5:9:1000 at 400 rpm for 20 min, with a pH of 6.0) at a fermentation temperature of 29° C. and a rotation speed of 200 rpm for 10 h to obtain a fermentation liquid;
[0072] (3) The fermentation broth was centrifuged at 4°C, 11,000 rpm for 10 min, and the supernatant was collected. 10% polyethylene glycol 6000 was first added to the supernatant at 4°C while stirring, and the mixture was allowed to stand at 4°C for 3 h. Then, 20% polyethylene glycol 6000 and 0.3 mol / L NaCl were added, and the mixture was allowed to stand at 4°C for 10 h. The mixture was then centrifuged at 12,000 rpm for 30 min, and the precipitate was collected and dissolved in a borate buffer solution with a pH of 9.0 to obtain a crude protease extract.
[0073] (4) The crude protease extract was placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa, and dialyzed with a pH 7.5, 20 mmol / L Tris-HCl buffer at 4°C to desalt the obtained protein solution. The protein solution was freeze-dried at -20°C for 2 h to obtain the microbial protease.
[0074] The Chinese medicine is prepared by the following method:
[0075] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 5:2.5:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 110-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.4:0.01, bacterial enzyme solution (prepared by mixing thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:2:3:8 at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 45°C for 6 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0076] In step S2, the functional agent is prepared by the following method:
[0077] 1) mannitol, vinyl pyrrolidone, water, and sodium percarbonate were stirred at 500 rpm for 20 minutes in a mass ratio of 16:8:50:2, sodium percarbonate was added, the temperature was controlled at 50° C., stirring was continued at 400 rpm for 5 hours under nitrogen protection, the mixture was cooled to room temperature, and centrifuged at 12,000 rpm for 10 minutes to obtain a functional compound;
[0078] 2) The functional substance was dispersed in deionized water (the amount was 50 times the mass of the functional substance) to form a functional substance solution. The mass ratio of the functional substance solution to the chitosan solution was 1.5:2. Chitosan solution (prepared by mixing chitosan and a 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 r / min for 30 min) was added to the functional substance solution. Ultrasonic dispersion was performed for 1 h. The ultrasonic power was controlled to 500 W and the ultrasonic frequency was controlled to 20 kHz. The mixture was centrifuged at 10,000 r / min for 30 min and then dried at 52° C. for 1.5 h to obtain the functional agent.
[0079] Example 4
[0080] A method for preparing collagen peptides comprises the following steps:
[0081] S1, by the mass ratio of fish scale powder, sodium hydroxide solution, citric acid solution is 1:10:6, the fish scales are crushed into fish scale powder and passed through an 80-mesh sieve, placed in a 5% sodium hydroxide solution with a mass concentration and soaked for 1 h, washed with water to neutrality, placed in a 5% citric acid solution with a mass concentration and soaked for 2.8 h after drying at 40 ° C, washed with water to neutrality, and dried for standby at 48 ° C;
[0082] S2, the obtained fish scale powder was added to the enzyme reactor, water was added and stirred at 400r / min for 30min, and then microbial protease and functional agent were added and stirred at 200r / min for 20min to carry out enzymatic hydrolysis at 48°C for 5.5h, and then the temperature was raised to 80°C to inactivate the enzyme for 18min to obtain a protease hydrolyzate;
[0083] S3. Centrifuge the protein hydrolysate at 8000 r / min for 10 min to obtain the supernatant, ultrafilter the supernatant using a 4.5 kDa ultrafiltration membrane with a controlled pressure of 0.55 MPa, and freeze-dry at -30°C for 9.5 h to obtain collagen peptides.
[0084] In step S2, the microbial protease is prepared by the following method:
[0085] (1) Activating Aspergillus oryzae was inoculated into a seed culture medium (prepared by mixing peptone, glucose, sodium chloride, and water in a mass ratio of 10:3:12:1000 at 400 rpm for 10 min, with a pH of 6.0), and culturing at 30°C for 11 h to obtain a seed solution;
[0086] (2) inoculating the seed liquid at an inoculum volume ratio of 4.5% into a fermentation medium (prepared by mixing Chinese medicinal materials, yeast powder, glucose, magnesium sulfate, and water in a mass ratio of 7.5:6.5:5:9.5:1000 at 400 rpm for 20 min, with a pH of 6.0) at a fermentation temperature of 29.5° C. and a rotation speed of 200 rpm for 10 h to obtain a fermentation liquid;
[0087] (3) The fermentation broth was centrifuged at 4°C, 11500 r / min for 10 min, and the supernatant was collected. 10% polyethylene glycol 6000 was first added to the supernatant at 4°C while stirring, and the mixture was allowed to stand at 4°C for 3 h. Then, 20% polyethylene glycol 6000 and 0.3 mol / L NaCl were added, and the mixture was allowed to stand at 4°C for 11 h. The mixture was then centrifuged at 12000 r / min for 30 min, and the precipitate was collected and dissolved in a borate buffer solution with a pH of 9.0 to obtain a crude protease extract.
[0088] (4) The crude protease extract was placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa, and dialyzed with a pH 7.5, 20 mmol / L Tris-HCl buffer at 4°C to desalt the obtained protein solution. The protein solution was freeze-dried at -20°C for 2 h to obtain the microbial protease.
[0089] The Chinese medicine is prepared by the following method:
[0090] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 5:2.8:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together to pass through a 115-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.45:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:2.5:3.5:8 and stirred and mixed at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 48°C for 6.5 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0091] In step S2, the functional agent is prepared by the following method:
[0092] 1) mannitol, vinyl pyrrolidone, water, and sodium percarbonate were stirred at 500 rpm for 20 minutes in a mass ratio of 18:8.5:50:2, sodium percarbonate was added, the temperature was controlled at 55° C., stirring was continued at 400 rpm for 5.5 hours under nitrogen protection, the mixture was cooled to room temperature, and centrifuged at 12,000 rpm for 10 minutes to obtain a functional compound;
[0093] 2) The functional substance was dispersed in deionized water (the amount was 50 times the mass of the functional substance) to form a functional substance solution. The mass ratio of the functional substance solution to the chitosan solution was 1.8:2. Chitosan solution (prepared by mixing chitosan and a 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 rpm for 30 minutes) was added to the functional substance solution. Ultrasonic dispersion was performed for 1 hour. The ultrasonic power was controlled to 550 W and the ultrasonic frequency was controlled to 20 kHz. The mixture was centrifuged at 10,000 rpm for 30 minutes and then dried at 56°C for 1.8 hours to obtain the functional agent.
[0094] Example 5
[0095] A method for preparing collagen peptides comprises the following steps:
[0096] S1, by the mass ratio of fish scale powder, sodium hydroxide solution, and citric acid solution is 1:10:6, the fish scales are crushed into fish scale powder and passed through an 80-mesh sieve, placed in a 5% sodium hydroxide solution with a mass concentration and soaked for 1 h, washed with water to neutrality, placed in a 5% citric acid solution with a mass concentration and soaked for 3 h after drying at 40 ° C, washed with water to neutrality, and dried for standby at 50 ° C;
[0097] S2, the obtained fish scale powder was added to the enzyme reactor, water was added and stirred at 400r / min for 30min, and then microbial protease and functional agent were added and stirred at 200r / min for 20min to perform enzymolysis. The enzymolysis temperature was 50°C, and the enzymolysis was performed for 6h. The temperature was raised to 80°C and the enzyme was inactivated for 20min to obtain a protease hydrolyzate;
[0098] S3. Centrifuge the protein hydrolysate at 8000 r / min for 10 min to obtain the supernatant, ultrafilter the supernatant using a 5 kDa ultrafiltration membrane, control the pressure at 0.6 MPa, and freeze-dry at -30°C for 10 h to obtain collagen peptides.
[0099] In step S2, the microbial protease is prepared by the following method:
[0100] (1) Activating Aspergillus oryzae was inoculated into a seed culture medium (prepared by mixing peptone, glucose, sodium chloride, and water in a mass ratio of 10:3:12:1000 at 400 rpm for 10 min, with a pH of 6.0), and culturing at 30°C for 12 h to obtain a seed solution;
[0101] (2) The seed liquid was inoculated at an inoculum volume ratio of 5% into a fermentation medium (prepared by mixing Chinese medicine, yeast powder, glucose, magnesium sulfate, and water in a mass ratio of 8:7:5:10:1000 at 400 rpm for 20 min, with a pH of 6.0) at a fermentation temperature of 30° C. and a rotation speed of 200 rpm for 10 h to obtain a fermentation liquid;
[0102] (3) The fermentation broth was centrifuged at 4°C, 12,000 rpm for 10 min, and the supernatant was collected. 10% polyethylene glycol 6000 was first added to the supernatant at 4°C while stirring, and the mixture was allowed to stand at 4°C for 3 h. Then, 20% polyethylene glycol 6000 and 0.3 mol / L NaCl were added, and the mixture was allowed to stand at 4°C for 12 h. The mixture was then centrifuged at 12,000 rpm for 30 min, and the precipitate was collected and dissolved in a borate buffer solution with a pH of 9.0 to obtain a crude protease extract.
[0103] (4) The crude protease extract was placed in a dialysis bag with a molecular weight cutoff of 3.5 kDa, and dialyzed with a pH 7.5, 20 mmol / L Tris-HCl buffer at 4°C to desalt the obtained protein solution. The protein solution was freeze-dried at -20°C for 2 h to obtain the microbial protease.
[0104] The Chinese medicine is prepared by the following method:
[0105] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 5:3:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 120-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.5:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:3:4:8 and stirring at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 50°C for 7 hours, and centrifuged at 10000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0106] In step S2, the functional agent is prepared by the following method:
[0107] 1) mannitol, vinyl pyrrolidone, water, and sodium percarbonate were stirred at 500 rpm for 20 minutes in a mass ratio of 20:9:50:2, sodium percarbonate was added, the temperature was controlled at 60° C., stirring was continued at 400 rpm for 6 hours under nitrogen protection, the mixture was cooled to room temperature, and centrifuged at 12,000 rpm for 10 minutes to obtain a functional compound;
[0108] 2) The functional substance was dispersed in deionized water (the amount was 50 times the mass of the functional substance) to form a functional substance solution. The mass ratio of the functional substance solution to the chitosan solution was 2:2. Chitosan solution (prepared by mixing chitosan and a 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 rpm for 30 minutes) was added to the functional substance solution. The mixture was ultrasonically dispersed for 1 hour. The ultrasonic power was controlled to 600 W and the ultrasonic frequency was controlled to 20 kHz. The mixture was centrifuged at 10,000 rpm for 30 minutes and then dried at 60°C for 2 hours to obtain the functional agent.
[0109] Comparative Example 1
[0110] Comparative Example 1 is the same as Example 1, except that Astragalus is used instead of Ginkgo biloba in the composition of the Chinese medicinal raw materials, as shown below:
[0111] The Chinese medicine is prepared by the following method:
[0112] According to the mass ratio of astragalus to ethanol solution of 7:20, the astragalus was crushed and passed through a 100-mesh sieve to obtain a traditional Chinese medicine powder, and the traditional Chinese medicine powder was stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin was 1:0.3:0.01, and the bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:1:2:8 and stirred at 300 r / min for 30 minutes) and bovine serum albumin were added to the mixed solution, and the mixture was stirred and mixed at 300 r / min for 20 minutes. The mixture was sealed and fermented at 40°C for 5 hours, and centrifuged at 10000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine.
[0113] Comparative Example 2
[0114] Comparative Example 2 is the same as Example 1, except that Ginkgo biloba is used instead of Astragalus membranaceus in the composition of the Chinese medicinal raw materials, as shown below:
[0115] The Chinese medicine is prepared by the following method:
[0116] According to the mass ratio of ginkgo leaf to ethanol solution of 7:20, the ginkgo leaf was crushed and passed through a 100-mesh sieve to obtain Chinese medicine powder, the Chinese medicine powder was stirred and mixed with 70% ethanol solution at 500 r / min for 30 min to obtain a mixed solution, according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin of 1:0.3:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase, bromelain and water at a mass ratio of 1:1:2:8, stirred and mixed at 300 r / min for 30 min) and bovine serum albumin were added to the mixed solution, stirred and mixed at 300 r / min for 20 min, sealed and fermented at 40°C for 5 h, and centrifuged at 10000 r / min for 10 min to remove the residue to obtain a Chinese medicine.
[0117] Comparative Example 3
[0118] Comparative Example 3 is the same as Example 1, except that the bacterial enzyme solution contains only thermophilic Streptococcus and amylase, as shown below:
[0119] The Chinese medicine is prepared by the following method:
[0120] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 2:5:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 100-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.3:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, amylase and water at a mass ratio of 1:3:8 and stirring at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 40°C for 5 hours, and centrifuged at 10000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0121] Comparative Example 4
[0122] Comparative Example 4 is the same as Example 1, except that the bacterial enzyme solution contains only thermophilic Streptococcus and bromelain, as shown below:
[0123] The Chinese medicine is prepared by the following method:
[0124] According to the mass ratio of astragalus, ginkgo leaf and ethanol solution being 2:5:20, the astragalus and ginkgo leaf are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 100-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, bacterial enzyme solution and bovine serum albumin being 1:0.3:0.01, bacterial enzyme solution (prepared by thermophilic streptococcus, bromelain and water at a mass ratio of 1:3:8 and stirring at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 40°C for 5 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0125] Comparative Example 5
[0126] Comparative Example 5 is the same as Example 1, except that only enzyme solution is used, as shown below:
[0127] The Chinese medicine is prepared by the following method:
[0128] According to the mass ratio of astragalus, ginkgo leaves and ethanol solution being 2:5:20, the astragalus and ginkgo leaves are stirred and mixed at 600 r / min for 10 minutes, and the mixture is pulverized together and passed through a 100-mesh sieve to obtain a traditional Chinese medicine powder, which is then stirred and mixed with a 70% ethanol solution at 500 r / min for 30 minutes to obtain a mixed solution, and according to the mass ratio of the mixed solution, enzyme solution and bovine serum albumin being 1:0.3:0.01, enzyme solution (prepared by mixing amylase, bromelain and water at a mass ratio of 1:2:8 at 300 r / min for 30 minutes) and bovine serum albumin are added to the mixed solution, and the mixture is stirred and mixed at 300 r / min for 20 minutes, sealed and fermented at 40°C for 5 hours, and centrifuged at 10,000 r / min for 10 minutes to remove the residue to obtain a traditional Chinese medicine agent.
[0129] Comparative Example 6
[0130] Comparative Example 6 is the same as Example 1, except that vinyl pyrrolidone is not grafted during the preparation of the functional agent, as shown below:
[0131] In step S2, the functional agent is prepared by the following method:
[0132] According to the mass ratio of mannitol to water being 12:50, mannitol and water were stirred and mixed at 500 r / min for 20 minutes to form a mannitol solution. According to the mass ratio of mannitol solution to chitosan solution being 1:2, chitosan solution (prepared by mixing chitosan and a dilute acetic acid solution with a mass concentration of 3% at a mass ratio of 1:10 at 400 r / min for 30 minutes) was added to the mannitol solution, and ultrasonic dispersion was performed for 1 hour, the ultrasonic power was controlled to 400 W, the ultrasonic frequency was controlled to 20 kHz, and then dried at 45° C. for 1 hour to obtain a functional agent.
[0133] Comparative Example 7
[0134] Comparative Example 7 is the same as Example 1, except that the functional agent is first coated with chitosan solution and then grafted with vinyl pyrrolidone, as shown below:
[0135] In step S2, the functional agent is prepared by the following method:
[0136] 1) mannitol and water were stirred at 500 r / min for 20 minutes in a mass ratio of mannitol to water of 12:50 to form a mannitol solution, chitosan solution (prepared by mixing chitosan and 3% dilute acetic acid solution at a mass ratio of 1:10 at 400 r / min for 30 minutes) was added to the mannitol solution in a mass ratio of mannitol to chitosan solution of 1:2, and ultrasonic dispersion was performed for 1 hour, with the ultrasonic power controlled at 400 W and the ultrasonic frequency controlled at 20 kHz, and then dried at 45° C. for 1 hour to obtain a functional material;
[0137] 2) The functional substance, vinyl pyrrolidone, water, and sodium percarbonate were stirred and mixed at 500 r / min for 20 minutes in a mass ratio of 12:7:50:2. Sodium percarbonate was added, the temperature was controlled at 40° C., and the mixture was stirred and mixed at 400 r / min for 4 hours under nitrogen protection. The mixture was cooled to room temperature and centrifuged at 12,000 r / min for 10 minutes to obtain a functional agent.
[0138] Comparative Example 8
[0139] Comparative Example 8 is the same as Example 1, except that the functional agent is not coated with chitosan solution during preparation, as shown below:
[0140] In step S2, the functional agent is prepared by the following method:
[0141] According to the mass ratio of mannitol, vinyl pyrrolidone, water and sodium percarbonate being 12:7:50:2, mannitol, vinyl pyrrolidone and water were stirred and mixed at 500 r / min for 20 minutes, sodium percarbonate was added, the temperature was controlled at 40°C, and the stirring and mixing was continued at 400 r / min for 4 hours under nitrogen protection. The mixture was cooled to room temperature and centrifuged at 12000 r / min for 10 minutes to obtain a functional agent.
[0142] Performance testing
[0143] Comprehensive performance tests were performed on the collagen peptides prepared in Examples 1-5 and Comparative Examples 1-8 of the present application.
[0144] 1. Determination of collagen peptide extraction rate and yield
[0145] The total nitrogen content of the collagen peptides prepared in Examples 1-5 and Comparative Examples 1-8, as well as the total nitrogen content of the corresponding raw fish scales, was determined according to the method in GB 5009.5-2016, "Determination of Protein in Foods." The collagen peptide extraction rate was calculated according to the following formula. The results are shown in Table 1.
[0146] Collagen peptide extraction rate (%) = (total nitrogen content of collagen peptides / total nitrogen content of fish scales) × 100%; collagen peptide yield (%) = (weight of collagen peptides / weight of fish scales) × 100%.
[0147] 2. Collagen Peptide Activity Assay
[0148] The antioxidant activity of the collagen peptides prepared in Examples 1-5 and Comparative Examples 1-8 was determined using an ABTS free radical scavenging ability kit and a superoxide anion free radical scavenging ability kit. The test results are shown in Table 1.
[0149] According to GB / T9695.23-2008, the hydroxyproline content of the collagen peptides prepared in Examples 1-5 and Comparative Examples 1-8 was tested. The results are shown in Table 1.
[0150] Table 1
[0151]
[0152]
[0153] As shown in Table 1, the extraction rate, yield and biological activity of the collagen peptides prepared in Examples 1-5 of the present application are significantly better than those prepared in Comparative Examples 1-8. Among them, Comparative Examples 1-5 are compared with Example 1 to show that changing the raw material composition and preparation process of the Chinese medicine can reduce the extraction efficiency, yield and activity of the collagen peptides, indicating that Astragalus and Ginkgo Biloba have the effect of synergistically improving the activity and extraction rate of the collagen peptides, and that thermophilic Streptococcus, amylase and bromelain are compounded to improve the yield and activity of Chinese medicine extraction rate and microbial protease, thereby improving the extraction efficiency and activity of the collagen peptides. Comparative Examples 6-8 are compared with Example 1 to show that changing the preparation process of the functional agent can significantly reduce the extraction efficiency and activity of the collagen peptides, indicating that mannitol is first grafted with vinyl pyrrolidone, and then coated with chitosan solution to enhance the binding force and affinity of the functional agent and collagen, thereby improving the extraction efficiency, extraction purity and biological activity of the collagen peptides.
[0154] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.
Claims
1. A method for preparing collagen peptide, characterized in that: The specific steps include the following: S1, fish scales are crushed into fish scale powder, placed in sodium hydroxide solution and soaked, washed with water to neutrality, then placed in citric acid solution and soaked, washed with water to neutrality, dried for standby use; S2, adding the fish scale powder obtained above to an enzyme reaction kettle, adding water and stirring to mix, then adding microbial protease and functional agent and stirring to mix, performing enzymatic hydrolysis, heating to inactivate the enzyme, and obtaining a protease hydrolyzate; S3. Centrifuge the protein hydrolysate to obtain the supernatant, ultrafilter the supernatant, and freeze-dry to obtain collagen peptides.
2. The method for preparing collagen peptide according to claim 1, wherein: In step S2, the microbial protease is prepared by the following method: (1) activating Aspergillus oryzae and inoculating the activated Aspergillus oryzae into a seed culture medium to obtain a seed solution; (2) inoculating the seed liquid into a fermentation medium and fermenting to obtain a fermentation liquid; (3) Centrifuging the fermentation broth, taking the supernatant, adding polyethylene glycol 6000 and NaCl to the supernatant at low temperature while stirring, letting it stand, and then centrifuging it, taking the precipitate, and dissolving the precipitate with borate buffer to obtain a crude protease extract; (4) The crude protease extract is placed in a dialysis bag, dialyzed with Tris-HCl buffer to remove salt to obtain a protein solution, and the protein solution is freeze-dried to obtain the microbial protease.
3. The method for preparing collagen peptide according to claim 2, wherein: In the step (2), the fermentation medium is prepared by the following method: Chinese medicinal agent, yeast powder, glucose, magnesium sulfate and water are stirred and mixed in a mass ratio of 6-8:5-7:5:8-10:1000.
4. The method for preparing collagen peptide according to claim 3, wherein: The Chinese medicine is prepared by the following method: mixing astragalus root and ginkgo leaves, crushing and sieving to obtain Chinese medicine powder, mixing the Chinese medicine powder with ethanol solution to obtain a mixed solution, adding bacterial enzyme solution and bovine serum albumin to the mixed solution, stirring and mixing, sealing and fermenting, and centrifuging to remove residue to obtain the Chinese medicine.
5. The method for preparing collagen peptide according to claim 4, characterized in that: The mass ratio of the astragalus root, ginkgo leaf and ethanol solution is 5:2-3:
20.
6. The method for preparing collagen peptide according to claim 4, characterized in that: The bacterial enzyme liquid is prepared by the following method: thermophilic streptococcus, amylase, bromelain and water are stirred and mixed in a mass ratio of 1:1-3:2-4:
8.
7. The method for preparing collagen peptide according to claim 1, characterized in that: In step S2, the functional agent is prepared by the following method: 1) Stirring mannitol, vinyl pyrrolidone and water, adding sodium percarbonate, heating, continuing stirring and mixing, cooling to room temperature, and centrifuging to obtain a functional substance; 2) The functional substance is dispersed in deionized water to form a functional substance solution, chitosan solution is added to the functional substance solution, ultrasonic dispersion is performed, and centrifugal drying is performed to obtain a functional agent.
8. The method for preparing collagen peptide according to claim 7, characterized in that: In the step 1), the mass ratio of mannitol, vinyl pyrrolidone, water and sodium percarbonate is 12-20:7-9:50:
2.
9. The method for preparing collagen peptide according to claim 7, characterized in that: In the step 2), the mass ratio of the functional substance solution to the chitosan solution is 1-2:
2.
10. A collagen peptide, characterized in that The collagen peptide is prepared by the preparation method of any one of claims 1 to 9.