Industrialized method for extracting epidermal growth factor (EGF) and protein polymerized by peptide from bovine colostrum permeate

Abstract

The invention relates to the technical field of bovine colostrum permeate extraction, in particular to an industrial method for extracting epidermal growth factors (EGF) from bovine colostrum permeate and protein polymerized by peptides. An industrial method for extracting protein polymerized by epidermal growth factors (EGF) and peptides from bovine coloctrum permeate comprises the following steps: (1) adjusting the pH value of the bovine coloctrum permeate to 4.3-5.1, and centrifuging to obtain a solid; (2) adding water into the solid for washing, centrifuging and drying to obtain protein powder polymerized from epidermal growth factors (EGF) and peptides; according to the present invention, the advanced process combining pH regulation and centrifugation is adopted, the efficient separation of the protein polymerized by the peptide and the epidermal growth factor EGF is achieved, the impurity interference is avoided, the component degradation possibly caused by the traditional extraction method is reduced, and the high quality and the high stability of the final product are ensured.

Description

Technical Field

[0001] The present application relates to the technical field of bovine colostrum permeate extraction, and in particular to an industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate. Background Art

[0002] Colostrum permeate, a byproduct of the colostrum ultrafiltration process, has traditionally been considered a waste product. However, colostrum contains significant amounts of lactose, soluble proteins, peptides, vitamins, and minerals, making it a nutrient-rich resource. Despite its high nutritional value, the disposal of colostrum permeate presents environmental concerns, leading to growing interest in its high-value-added utilization. Colostrum permeate retains over 70% of the lactose found in milk, along with varying proportions of other components. However, direct application of colostrum permeate in the food industry is limited due to lactose's low sweetness, poor solubility, and difficulty in digestion for those with lactose intolerance.

[0003] Currently, the utilization of bovine colostrum permeate faces significant economic and technical challenges. In existing technologies, bovine colostrum permeate is hydrolyzed and fermented through lactose to produce sports drinks. However, the high cost of enzyme preparations, large equipment investment, low process efficiency and strict process control requirements make large-scale production difficult.

[0004] Therefore, how to directly extract epidermal growth factor EGF and peptide polymerized proteins from dairy by-products, reduce the generation of waste during the extraction process, and maximize the utilization of all residual components has become an urgent problem to be solved. Summary of the Invention

[0005] In response to the shortcomings of the existing technology, the present application provides an industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized proteins from bovine colostrum permeate. The present application proposes an innovative and sustainable method for extracting bioactive peptide-polymerized proteins and epidermal growth factor (EGF) directly from dairy by-products such as bovine colostrum permeate. This new method does not require the use of enzymes, heating, microbial activity, or other high-energy or resource-intensive processes, thereby ensuring that the biological activity of the peptides is retained. By maximizing the utilization of all residual components, the present application not only reduces waste but also meets the goals of sustainable development.

[0006] This breakthrough technology transforms a once-waste dairy byproduct into a high-value raw material, providing the dairy industry with a cost-effective and environmentally friendly solution. By unlocking the potential of bovine colostrum permeate, this approach paves the way for a more efficient, sustainable, and profitable future for dairy processing.

[0007] In a first aspect, the present application provides an industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized proteins from bovine colostrum permeate, using the following technical solution: An industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate, the method comprising the following steps: (1) adjusting the pH of the colostrum permeate to 4.3-5.1, centrifuging, and obtaining a solid; (2) Water is added to the solid matter for washing, centrifugation, and drying to obtain a powder containing epidermal growth factor EGF and a peptide-polymerized protein.

[0008] By adopting the above technical solution, this application proposes an innovative and sustainable method, which uses an advanced process combining pH adjustment, centrifugation and pure water washing to directly extract proteins and epidermal growth factor (EGF) polymerized from bioactive peptides from the permeate of bovine colostrum, a dairy by-product, to achieve efficient separation of proteins polymerized from peptides and epidermal growth factor EGF.

[0009] This application discloses an industrialized method for extracting a protein composed of epidermal growth factor (EGF) and peptides from bovine colostrum permeate, offering both process advantages and safety. This method, without the addition of any preservatives or chemical additives, relies solely on pH adjustment and centrifugal separation, ensuring the naturalness and safety of the product. This pH adjustment and physical separation maximize the biological activity of the peptide and EGF, while avoiding the potential negative effects of high temperatures or chemical treatments.

[0010] When bovine colostrum permeate is left to stand, the microorganisms in the permeate will continue to ferment and produce acid, causing the pH to drop continuously (approximately 0.3-0.5 pH units / hour). This may also affect the stability of heat-sensitive ingredients (such as active peptides and epidermal growth factor (EGF)). Therefore, when adjusting the pH, control the pH to 4.3-5.1 and complete subsequent steps as quickly as possible to avoid excessive acidification or microbial metabolites that affect the activity and yield of the target ingredients. If extended processing time is required, consider temporary storage at low temperature (4-10°C) or adding an appropriate amount of antibacterial agent (such as 0.02-0.05% potassium sorbate) to slow the fermentation process.

[0011] Preferably, the bovine colostrum permeate in step (1) is a bovine colostrum permeate produced by ultrafiltration of defatted bovine colostrum.

[0012] Preferably, the method for preparing the defatted bovine colostrum comprises the following steps: defatting the bovine colostrum by centrifugation to obtain the defatted bovine colostrum.

[0013] Preferably, the bovine colostrum permeate in step (1) is the bovine colostrum permeate produced by ultrafiltration of bovine colostrum.

[0014] Preferably, in step (1), the pH of the bovine colostrum permeate is adjusted to 4.4-4.6.

[0015] Preferably, the reagent used to adjust the pH of the bovine colostrum permeate is an acidic reagent.

[0016] Preferably, the acidic agent includes at least one of citric acid, vitamin C or acetic acid.

[0017] Preferably, the centrifugal speed in step (1) is 5000-8000 rpm, and the centrifugal time is 15-25 minutes.

[0018] Preferably, the centrifugal speed in step (2) is 5000-8000 rpm, and the centrifugal time is 15-25 minutes.

[0019] By adopting the above technical solution, in this application, the pH of the bovine colostrum permeate is adjusted to 4.3-5.1, the mixture is centrifuged for the first time, the lactose in the supernatant is removed, and the precipitate is retained; then, pure water is added to the precipitate for washing, and after thorough mixing, centrifugation is performed again. After the second centrifugation, the supernatant is removed, and finally a powder containing epidermal growth factor EGF and a peptide-polymerized protein is obtained.

[0020] This application achieves efficient separation through two centrifugations and pure water washing to ensure the purity and activity of the target product.

[0021] Preferably, the drying method includes freeze drying, spray drying and other drying methods.

[0022] In a second aspect, the present application provides a powder containing epidermal growth factor (EGF) and a protein polymerized from peptides, using the following technical solution: A powder containing epidermal growth factor EGF and a protein polymerized from peptides is prepared by the above method.

[0023] Preferably, the content of epidermal growth factor (EGF) in the powder is 1600-2200 ng / g; and the content of protein synthesized by peptide polymerization is 60-76 wt%.

[0024] In a third aspect, the present application provides a method for preparing a food, health product or medicine by using a powder containing epidermal growth factor (EGF) and a protein synthesized by peptide polymerization, using the following technical solution: The powder containing the protein polymerized from epidermal growth factor EGF and peptides is used in the preparation of food, health products or medicines.

[0025] The prepared protein powder containing epidermal growth factor EGF and peptide polymerization can be directly developed into liquid beverage.

[0026] Water is added to the solid matter for washing, and the precipitate obtained after centrifugation can be directly used to develop liquid beverages without going through a drying step.

[0027] In summary, this application includes at least one of the following beneficial technical effects: 1. This application discloses an industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized proteins from bovine colostrum permeate. This application uses an advanced process combining pH adjustment and centrifugation to achieve efficient separation of peptide-polymerized proteins and epidermal growth factor (EGF), avoiding interference from impurities. At the same time, this process reduces the degradation of components that may be caused by traditional extraction methods, ensuring the high quality and stability of the final product. 2. This application uses a purely physical method to directly extract highly active biological components from bovine colostrum permeate, without adding any enzymes, microorganisms, preservatives, fermentation agents or any chemical reagents, thus avoiding chemical residues and biological contamination and ensuring the safety and purity of the extract; 3. The process of this application is simple and suitable for industrial application; the production process of this application is reasonable, the steps are concise, and it avoids complex chemical reactions and tedious operations, making it easy to apply on a large scale in industrial applications; 4. The present application makes full use of the bovine colostrum permeate, a by-product of traditional dairy processing, and converts it into high-value-added biological products, thereby realizing efficient recycling of resources and reducing environmental pollution problems, in line with the concepts of circular economy and sustainable development; 5. The present application discloses an industrial method for extracting epidermal growth factor EGF and peptide-polymerized proteins from bovine colostrum permeate. The extracted epidermal growth factor EGF and peptide-polymerized proteins have high biological activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a production process flow chart for extracting epidermal growth factor (EGF) and peptide-polymerized proteins from bovine colostrum permeate; Figure 2 This is the SDS-PAGE electrophoresis analysis of the permeate powder, permeate supernatant powder, and powder samples containing epidermal growth factor EGF and peptide-polymerized protein. Lanes 1, 3, and 6 are powders containing epidermal growth factor EGF and peptide-polymerized protein; lanes 2, 4, and 8 are permeate powder; and lanes 5 and 7 are permeate supernatant powder. Figure 3 These are infrared spectra of permeate powder, permeate supernatant powder, and powder sample containing epidermal growth factor EGF and peptide-polymerized protein; Figure 4Results of the antioxidant ABTS free radical scavenging experiment of the permeate powder, the permeate supernatant powder, and the powder containing the protein synthesized from epidermal growth factor EGF and peptides. DETAILED DESCRIPTION

[0029] The technical solution of the present application is further illustrated below through specific embodiments. The specific embodiments do not limit the scope of protection of the present application; some non-essential modifications and adjustments made by others based on the concept of the present application still fall within the scope of protection of the present application.

[0030] The raw materials involved in this application are all commercially available products, and the application is further described in detail below in conjunction with the examples and comparative examples.

[0031] The bovine colostrum permeate in the present application can be bovine colostrum permeate or defatted bovine colostrum permeate. The bovine colostrum permeate and defatted bovine colostrum permeate can be commercially available permeates, or permeates obtained through laboratories, factories, etc. In this embodiment, the bovine colostrum permeate is obtained from factory production.

[0032] The preparation method of bovine colostrum permeate in the embodiment of the present application comprises the following steps: Step 1: Collect colostrum: milk the cows and collect the colostrum, and control the temperature of the colostrum to 4-18°C; Step 2: Centrifugal degreasing: transport the bovine colostrum to a centrifugal degreasing machine for degreasing to obtain defatted bovine colostrum; The current setting value of the centrifugal degreasing machine is 7.5-8.5A, and the speed is 3000-6000rpm; during the centrifugal degreasing process, the temperature of the bovine colostrum is controlled to be ≤25℃; Step 3, ultrafiltration and concentration: ultrafiltration and concentration of the defatted bovine colostrum to obtain a concentrate and a bovine colostrum permeate waste; Wherein, during ultrafiltration concentration, ultrafiltration equipment with a membrane molecular weight cut-off of ≤20KDa is used for ultrafiltration concentration.

[0033] The present application provides an industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate, the specific steps of which are as follows: An industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate, the method comprising the following steps: (1) Collection of bovine colostrum permeate waste: The colostrum permeate waste after ultrafiltration is collected and stored in a dedicated storage tank for subsequent use.

[0034] (2) Remove lactose: The pH of the milk primary permeate is adjusted using an acidic reagent to precipitate the peptides and growth factors, and the supernatant is removed by centrifugation to obtain a precipitate.

[0035] The specific steps are as follows: The pH of the bovine colostrum permeate is adjusted to 4.3-5.1 with an acidic reagent, and then centrifuged to remove lactose in the supernatant to obtain a solid.

[0036] More preferably, the pH of the bovine colostrum permeate is adjusted to 4.4-4.6.

[0037] Preferably, the centrifugal speed in step (1) is 5000-8000 rpm, and the centrifugal time is 15-25 minutes.

[0038] Preferably, the acidic agent includes at least one of citric acid, vitamin C or acetic acid.

[0039] In a specific embodiment, the specific steps for removing lactose are as follows: the pH value of the bovine colostrum permeate is adjusted by adding an acidic reagent (food-grade acidic reagent). The acidic reagent (food-grade acidic reagent) can be added directly in powder form or first dissolved in purified water to prepare a solution and then added; under low pH conditions, the peptide and protein combine to form a precipitate, which is centrifuged to obtain a solid material containing a large amount of high-purity low-molecular-weight protein.

[0040] (3) Washing, centrifugation and drying steps: The obtained solid matter was added with pure water and centrifuged for washing, centrifuged and dried to obtain a powder containing a protein polymerized from epidermal growth factor EGF and the peptide.

[0041] The specific steps are as follows: Water is added to the solid matter for washing, centrifugation, and drying to obtain a powder containing epidermal growth factor EGF and a protein polymerized from the peptide.

[0042] In the washing step, pure water is added to the obtained solid matter, and after thorough mixing, the solid matter is centrifuged again to further remove impurities, thereby finally obtaining a powder containing epidermal growth factor EGF and a protein polymerized from the peptide.

[0043] Preferably, the centrifugal speed in step (2) is 5000-8000 rpm, and the centrifugal time is 15-25 minutes.

[0044] The drying method includes freeze drying, spray drying and other drying methods.

[0045] The present invention will be further described below by way of examples.

[0046] Example 1: An industrial method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate, the process flow is as follows: Figure 1As shown, the following steps are included: (1) Collecting bovine colostrum: Milk the cows and collect the colostrum, which is immediately refrigerated at 4°C. (2) Centrifugal degreasing: The colostrum is transported to a centrifugal degreasing machine for degreasing to obtain defatted colostrum; The current setting value of the centrifugal skimmer is 8.2A, the speed is 5000rpm, and the initial temperature of the milk is controlled to be ≤25℃ during the centrifugal skimming process; (3) Ultrafiltration concentration: Ultrafiltration and concentration of the defatted bovine colostrum are performed to obtain a concentrate and a bovine colostrum permeate (i.e., defatted bovine colostrum permeate); Wherein, during ultrafiltration concentration, ultrafiltration equipment with a membrane molecular weight cut-off of 20KDa is used for ultrafiltration concentration.

[0047] (4) Collecting bovine colostrum permeate: The colostrum permeate is collected and further processed under low pH conditions or during pH adjustment.

[0048] (5) Remove lactose Dissolve food-grade citric acid in purified water to prepare a 2 M citric acid solution.

[0049] 2M citric acid solution was gradually added to 100 kg of bovine colostrum permeate with continuous stirring. The pH of the bovine colostrum permeate was adjusted to 4.6. Under low pH conditions, peptides and proteins combined to form a precipitate. The precipitate was centrifuged at 6500 rpm for 20 minutes, and the supernatant was removed by centrifugation to obtain a solid.

[0050] The SDS-PAGE test results of the solid matter are as follows Figure 2 As shown in the figure, it shows that the solid material contains a large amount of high-purity small molecular weight proteins.

[0051] (6) Washing, centrifugation, and drying steps In the washing step, pure water is added to the obtained solid matter, and after thorough mixing, the solid matter is centrifuged again at 6500 rpm for 20 minutes to further remove impurities and dried to obtain a powder containing epidermal growth factor EGF and a peptide-polymerized protein.

[0052] This example uses freeze-drying as an example, and the freeze-drying time is 35 hours.

[0053] Performance testing: The components and properties of the prepared bovine colostrum permeate, permeate supernatant, and powder containing epidermal growth factor (EGF) and peptide-polymerized protein were tested.

[0054] The bovine colostrum permeate and the permeate supernatant are dried to obtain a powder sample. The specific steps are as follows: The bovine colostrum permeate (prepared in step 4 of Example 1) was freeze-dried for 35 hours to obtain a permeate powder.

[0055] The permeate supernatant (ie, the supernatant after centrifugation, prepared in step 5 of Example 1) was freeze-dried for 35 hours to obtain a permeate supernatant powder.

[0056] 1. Quantitative analysis of sample components The components of the permeate powder, the permeate supernatant powder, and the powder containing epidermal growth factor (EGF) and peptide-polymerized protein prepared in Example 1 were tested, and the test results are as follows: Table 1 Quantitative analysis of the components of the permeate powder, the permeate supernatant powder, and the powder sample containing the protein synthesized from epidermal growth factor EGF and peptides Table 1 shows the results of quantitative analysis of the components of the peptide powder sample extracted from the permeate powder, the permeate supernatant powder, and the powder sample containing epidermal growth factor (EGF) and peptide-polymerized proteins. The permeate powder had a lactose content of 67.50 wt%, a protein content of 6.01 wt%, an ash content of 12.50 wt%, and an EGF content of 297.41 ng / g, indicating a high lactose content but relatively low protein and EGF contents. The permeate supernatant powder had a lactose content of 72.50 wt%, a protein content of 4.50 wt%, an ash content of 10.50 wt%, and an EGF content of 102.06 ng / g, indicating a further increase in lactose content but a significant decrease in protein and EGF contents. In contrast, the powder containing epidermal growth factor EGF and peptide-polymerized protein has a lactose content of only 4.39wt%, a protein content of up to 75.69wt%, an ash content of 11.92wt%, and an EGF content of 2126.58ng / g, showing an obvious peptide and growth factor concentration effect.

[0057] The protein and EGF content in the powder containing epidermal growth factor (EGF) and peptide-polymerized proteins was significantly higher than that in the permeate powder and permeate supernatant powder, demonstrating that the separation and purification process can efficiently concentrate peptides and growth factors. These peptides are derived from small molecule immune peptides, casein peptides, whey protein peptides, lactoferrin peptides, growth factor peptides, β-lactoglobulin peptides, α-lactalbumin peptides, antimicrobial peptides, and proline peptides, and possess diverse biological activities such as enhancing immunity, anti-oxidation, anti-inflammation, and promoting tissue repair.

[0058] As shown in Table 1, the EGF content in the powder containing EGF and peptide-polymerized protein is significantly higher than that in the permeate powder and permeate supernatant powder, indicating its higher nutritional value. EGF is a key growth factor that plays an important role in promoting cell growth, tissue repair, and enhancing skin elasticity. Therefore, the powder containing EGF and peptide-polymerized protein is not only a high-purity peptide source but also a functional ingredient rich in growth factors, with extremely high nutritional and health value.

[0059] This application discloses an industrialized method for extracting a protein composed of epidermal growth factor (EGF) and peptides from bovine colostrum permeate, offering both process advantages and safety. This method, without the addition of any preservatives or chemical additives, relies solely on pH adjustment and centrifugation, ensuring the naturalness and safety of the product. This pH adjustment and physical separation maximize the biological activity of the peptides and EGF, avoiding the potential negative effects of high temperatures or chemical treatments. Furthermore, strict sanitary conditions are maintained throughout the entire process to ensure the safety and purity of the final product.

[0060] The results in Table 1 demonstrate the significant advantages of a powder containing epidermal growth factor (EGF) and peptide-polymerized proteins in terms of peptide and growth factor content. Through innovative processing of the colostrum permeate, key components such as peptides and EGF are concentrated to the greatest extent possible, demonstrating their unique nutritional advantages. Conventional dairy processing methods often fail to effectively retain high-purity peptides and growth factors. However, this method, through the efficient separation and purification of the colostrum permeate, allows for the efficient concentration and retention of these key components, ensuring their superior nutritional value.

[0061] 2. SDS-PAGE electrophoresis analysis of samples: The results of SDS-PAGE electrophoresis analysis of powder samples are as follows: Figure 2 Lanes 1, 3, and 6 represent powders containing proteins aggregated from epidermal growth factor (EGF) and peptides; lanes 2, 4, and 8 represent permeate powders; and lanes 5 and 7 represent permeate supernatant powders. Peptide and EGF bands are clearly visible in the powders containing proteins aggregated from epidermal growth factor (EGF) and peptides (lanes 1, 3, and 6), particularly at molecular weights below 85 kDa, 75 kDa, 70 kDa, 33 kDa, 30 kDa, 35 kDa, 10 kDa, and 8 kDa, showing high concentrations of peptide components, including EGF. These low-molecular-weight peptides form precipitates by binding to high-molecular-weight substances, demonstrating the efficient concentration of peptides by this method. The permeate powders (lanes 2, 4, and 8) and permeate supernatant powders (lanes 5 and 7) exhibit almost no distinct peptide bands, indicating low peptide content in these samples.

[0062] Peptide content testing revealed that the peptide content of the powder containing epidermal growth factor (EGF) and peptide-polymerized proteins prepared using the method of this application was as high as 60%, demonstrating the method's remarkable effectiveness in peptide extraction and concentration. This technology, through pH adjustment and centrifugal separation, eliminates the need for preservatives or chemical additives, ensuring the extracted peptides possess high biological activity while maintaining their naturalness and safety. This pH adjustment and physical separation process not only effectively removes lactose but also maximizes the preservation of the peptide's structure and function.

[0063] This new technology not only offers advantages in peptide extraction and concentration but also has broad potential for application. By utilizing waste materials and by-products, this technology can provide raw materials for the development of a variety of products, particularly in the fields of food, health supplements, and pharmaceuticals. The protein powder containing epidermal growth factor (EGF) and peptides exhibits significant biological activity in enhancing immunity, providing antioxidant and anti-inflammatory benefits, and promoting tissue repair, giving it great potential for application in medicine and scientific research. The various peptide products developed using this technology could provide innovative solutions for the food industry, nutritional supplements, and even medical treatments.

[0064] Furthermore, the high-purity protein powder containing epidermal growth factor (EGF) and peptides extracted using this technology also possesses significant functionality and can be widely used in a variety of industries, including food processing and medical research, providing a new approach for the high-value utilization of waste materials. Through innovative extraction and concentration methods, resources typically considered waste can be transformed into highly value-added bioactive ingredients, bringing economic benefits to various industries while also providing innovative solutions for sustainable development.

[0065] 3. Infrared spectra of permeate powder, permeate supernatant powder and powder containing epidermal growth factor EGF and peptide polymerized protein. Infrared spectra of permeate powder, permeate supernatant powder and powder containing epidermal growth factor EGF and peptide polymerized protein. Figure 3 It can be seen that the infrared spectra of the permeate powder and the permeate supernatant powder show similar chemical compositions, mainly indicating that they contain high lactose components, with the lactose content exceeding 65%. In the spectrum, 1034 cm -1 and 894cm -1 The absorption peak of 1655cm is very obvious, representing the characteristic absorption of lactose. In addition, the spectrum also shows the absorption peak of 1655cm -1 and 1546cm -1 These peaks correspond to amide I band (C=O stretching vibration) and CN stretching vibration, indicating that these two samples also contain a small amount of peptides or proteins, but their intensities are weak, indicating that the peptide content is relatively low, about 3%.

[0066] The infrared spectrum of the powder containing epidermal growth factor (EGF) and peptide-polymerized protein showed significantly different characteristics compared with the permeate powder and the permeate supernatant powder. The absorption peak related to lactose (1034 cm -1 ) completely disappears from the spectrum of the powder containing epidermal growth factor (EGF) and peptide-polymerized proteins, indicating that lactose was successfully removed during the preparation process. This result demonstrates that lactose removal is achieved through pH adjustment and centrifugation. These treatments avoid the use of high temperatures, enzymes, or fermentation, effectively protecting the proteins and other active ingredients in the sample. pH adjustment facilitates the separation of lactose from other components, while centrifugation helps remove lactose and impurities while protecting the proteins and other active ingredients, ensuring the quality and functional properties of the sample.

[0067] In the spectrum of a powder containing a protein polymerized from epidermal growth factor (EGF) and peptides, the peak at 1655 cm -1 and 1546cm -1 The absorption peak of amide I is obvious and is at 1238 cm -1 The absorption peak of CN stretching vibration appears at , indicating that the sample has a high peptide or protein content. Because no high temperature or enzymes are used during pH adjustment and centrifugation, the protein structure is effectively protected, and aggregation occurs during the treatment, forming a more concentrated protein structure.

[0068] 4. Scavenging activity of permeate powder, permeate supernatant powder, and powder samples containing epidermal growth factor EGF and peptide-polymerized protein on ABTS free radicals at different concentrations. Scavenging activity of permeate powder, permeate supernatant powder, and powder samples containing epidermal growth factor EGF and peptide-polymerized protein on ABTS free radicals at different concentrations. Figure 4As shown, the antioxidant capacity of each sample was evaluated by comparing its scavenging activity. The concentration range was 3.0 mg / mL to 9.0 mg / mL. The results showed that the powder containing EGF and peptide-polymerized protein exhibited high ABTS free radical scavenging activity at all concentrations, particularly at 9.0 mg / mL, where the scavenging rate reached approximately 77.30%, significantly higher than that of the permeate powder (approximately 31.19%) and permeate supernatant powder (approximately 31.90%). This phenomenon indicates that the powder containing EGF and peptide-polymerized protein exhibits significant ability to combat ABTS free radicals. The permeate powder and permeate supernatant powder were relatively ineffective in scavenging ABTS free radicals, with maximum scavenging rates not exceeding 32%. This may be related to their compositional characteristics, particularly the high protein content in bovine colostrum and the high lactose content in the permeate waste, which to some extent inhibit the concentration and activity of antioxidant enzymes. In contrast, the powder containing EGF and peptide-polymerized protein is processed to be enriched with lysozyme and lactoperoxidase, enzymes that play an important role in scavenging free radicals. The unique processing of the powder containing EGF and peptide-polymerized protein effectively removes interfering substances such as lactose and excess protein, significantly increasing the concentration of antioxidant compounds, resulting in higher activity in scavenging ABTS free radicals. ABTS free radical scavenging experiments demonstrate that the powder containing EGF and peptide-polymerized protein exhibits significant antioxidant capacity. It demonstrates excellent potential in removing harmful free radicals and preventing the damage caused by oxidative stress. This antioxidant property makes the powder containing EGF and peptide-polymerized protein not only a nutritional supplement but also provides effective antioxidant protection, making it particularly suitable for use as an ingredient in health promotion and functional food development.

[0069] Example 2: In order to study the effect of different pH values ​​on the components of the powder containing epidermal growth factor (EGF) and peptide-polymerized protein, the bovine colostrum permeate in step (5) of Example 1 was adjusted to different pH values, and the contents of the various components in the prepared powder samples containing epidermal growth factor (EGF) and peptide-polymerized protein were detected, as shown in Table 2.

[0070] Table 2 Effect of pH on the composition of samples The results in Table 2 show that in the industrial extraction of EGF and peptide-polymerized proteins from bovine colostrum permeate, pH control plays a crucial role in the extraction efficiency and product quality of key components. The core goal is to maximize the extraction of EGF and peptide-polymerized proteins while simultaneously removing high-purity lactose, thereby enhancing the purity and functionality of the final protein product. As a functional protein, EGF reaches its highest extraction efficiency of 2126.58 ng / g at pH 4.6, the optimal pH for industrial EGF extraction. At this pH, EGF's structure is stable, facilitating dissolution and extraction, ensuring efficient recovery and preserving its biological activity. Further increases in pH to 5.0 and 5.1 result in EGF content decreasing to 1816.20 ng / g and 1667.59 ng / g, respectively, indicating that EGF may degrade or become inactivated in a slightly alkaline environment. Therefore, controlling the pH to 4.6 is most conducive to efficient EGF extraction and is suitable for industrial-scale production. The protein content also reached a peak of 75.69wt% at pH 4.6, indicating that under these conditions the protein structure is suitable for hydrolysis, the peptides are fully released, and it has good polymerization ability, which is conducive to the formation of high-molecular-weight, stable protein aggregates. At pH 5.1, the protein content gradually decreased to 61.75wt%, which may be due to further protein degradation or peptide instability, affecting the protein polymerization effect. Therefore, pH 4.6 is not only a key window for peptide release, but also an ideal condition for protein polymerization, which helps to prepare high-purity, high-functionality protein products that meet industrial standards.

[0071] Lactose is a waste product of bovine colostrum permeate, and its removal efficiency is also affected by pH. Data show that the lactose content increases with increasing pH, from 2.09wt% to 18.45wt%. At pH 4.6, the lactose content is as low as 4.39wt%, which is a favorable condition for effective lactose removal, helping to improve the purity of protein products and avoid lactose residues. Continuing to increase the pH to 5.0 and above, the lactose content rises sharply, indicating that lactose release intensifies, which is not conducive to lactose removal and increases the burden of subsequent processing. Therefore, controlling the pH at 4.6 can not only optimize the extraction and polymerization of proteins and peptides, but also minimize the interference of lactose, which is conducive to the production of high-purity, low-impurity protein products.

[0072] The ash content dropped to 11.92wt% at pH 4.6 and further to 9.8wt% at pH 5.1, indicating increased mineral dissolution. This helps to adjust the proportion of inorganic components in the product to a certain extent, but the ash content needs to be balanced in industry to ensure that the product meets functional protein standards. Comprehensive analysis shows that pH 4.6 is the optimal condition for extracting EGF and peptides from bovine colostrum permeate, achieving protein polymerization, and removing lactose. At this pH, the extraction efficiency of EGF and peptides is the highest, the lactose content is the lowest, and the protein polymerization effect is the best, forming a high-purity, highly functional protein product suitable for industrial production. Through precise pH control, the extraction efficiency can be effectively improved, the subsequent high-purity lactose extraction process can be simplified, energy consumption and costs can be reduced, and stable technical support can be provided for large-scale industrial applications, promoting the high-value utilization of bovine colostrum.

[0073] The present application discloses an industrialized method for extracting epidermal growth factor (EGF) and a peptide-polymerized protein from bovine colostrum permeate. The present application adopts an advanced process combining pH adjustment, centrifugation and pure water washing to achieve efficient separation of peptides and growth factor EGF, avoid interference from impurities, optimize the purification process, increase the recovery rate of the target product to more than 1.34%, and ensure that the protein purity is stably maintained at more than 70%; at the same time, the process reduces the degradation of components that may be caused by traditional extraction methods, ensuring the high quality and stability of the final product.

[0074] This application uses a purely physical method to directly extract highly active biological components from bovine colostrum permeate, without the addition of enzymes, microorganisms, preservatives, fermentation agents, or any other chemical reagents. This process avoids chemical residues and biological contamination, ensuring the safety and purity of the extract. This green extraction technology preserves the natural properties of the bioactive substances and meets the high demands of modern consumers for health products.

[0075] The production process presented in this application is well-designed, with simple steps, avoiding complex chemical reactions and tedious operations, making it amenable to large-scale industrial application. This technology can be retrofitted onto existing production lines, reducing reliance on high-end equipment and thus lowering both equipment investment and technological barriers to entry. Furthermore, this method utilizes low-cost processes to produce high-value products, improving economic efficiency and market competitiveness, and providing a sustainable development solution for enterprises.

[0076] This application fully utilizes colostrum permeate, a byproduct of traditional dairy processing, by converting it into high-value-added bioproducts, achieving efficient resource recycling. Compared to traditional treatment methods such as direct discharge or low-value utilization, this technology improves the utilization rate of waste resources while reducing environmental pollution, in line with the concepts of circular economy and sustainable development.

[0077] The products prepared using this application have highly active functional ingredients that meet health needs. The extracted epidermal growth factor (EGF) and the protein synthesized from peptides both have high biological activity, can promote cell repair and tissue regeneration, and meet the human body's need for functional ingredients. Research has shown that these bioactive substances have broad application value in health fields such as skin care, immune regulation, and anti-aging. At the same time, they can serve as high-quality nutritional supplements, providing reliable support for the research and development of health foods, medicines, and functional skin care products.

[0078] This application is environmentally friendly and energy-efficient, reducing production costs. The process does not require high-energy-consuming equipment and does not involve high temperatures, high pressures, or other process conditions, reducing energy consumption and overall production costs. Compared to traditional chemical or biological fermentation methods, this technology reduces carbon emissions while also avoiding the use of additional chemical reagents and catalysts, ensuring a more environmentally friendly production process that complies with global sustainable development policies.

[0079] This application has broad applicability and can expand product development. The protein synthesized from the high-purity epidermal growth factor (EGF) and peptides obtained through this method can be used as a core functional ingredient in a wide range of industries. In addition to directly developing high-value-added end products (such as health foods, functional cosmetics, and pharmaceuticals), it can also be used as an additive in a variety of products such as food, health products, and pharmaceutical preparations, further expanding market space and enhancing product diversification and competitiveness.

[0080] The products prepared in this application have broad market prospects and significant technical advantages. Due to their significant advantages in extraction technology, resource utilization, biological activity, environmental protection and energy conservation, this technology has attracted great attention from research institutions, production companies, and the consumer market. As global demand for natural, safe, and effective health products continues to grow, this innovative technology has enormous market potential, not only promoting the development of the bioextraction industry but also creating new business opportunities for companies.

Claims

1. An industrial process for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate, characterized by: The steps of the method are as follows: (1) Adjusting the pH of the colostrum permeate to 4.3-5.1, centrifuging, and obtaining a solid; (2) Water is added to the solid matter for washing, centrifugation, and drying to obtain a powder containing epidermal growth factor EGF and peptide-polymerized protein.

2. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 1, characterized in that: The bovine colostrum permeate in step (1) is the bovine colostrum permeate produced by ultrafiltration of defatted bovine colostrum.

3. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 1, characterized in that: The bovine colostrum permeate in step (1) is the bovine colostrum permeate produced by ultrafiltration of bovine colostrum.

4. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 1, characterized in that: In the step (1), the pH of the bovine colostrum permeate is adjusted to 4.4-4.

6.

5. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 4, characterized in that: The reagent used to adjust the pH of the bovine colostrum permeate is an acidic reagent.

6. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 5, characterized in that: The acidic agent includes at least one of citric acid, vitamin C or acetic acid.

7. The industrialized method for extracting epidermal growth factor (EGF) and peptide-polymerized protein from bovine colostrum permeate according to claim 1, characterized in that: In step (1), the centrifugal speed is 5000-8000 rpm, and the centrifugal time is 15-25 minutes; The centrifugal speed in step (2) is 5000-8000 rpm, and the centrifugal time is 15-25 minutes.

8. A powder containing epidermal growth factor (EGF) and a peptide-polymerized protein, characterized in that: The powder containing the protein polymerized from epidermal growth factor EGF and peptides is prepared by the method according to any one of claims 1 to 7.

9. The powder containing protein polymerized from epidermal growth factor (EGF) and peptides according to claim 8, characterized in that: The content of epidermal growth factor (EGF) in the powder is 1600-2200 ng / g; the content of protein synthesized by peptide polymerization is 60-76 wt %.

10. Use of the protein powder containing epidermal growth factor (EGF) and peptides according to claim 8 or 9 in the preparation of foods, health products or medicines.

Images