Preparation and purification process of fresh meat polypeptide, polypeptide composition and application of polypeptide composition

Abstract

The invention relates to the technical field of polypeptides, in particular to a preparation and purification process of fresh meat polypeptides, a polypeptide composition and application of the polypeptide composition, and the preparation and purification process of the fresh meat polypeptides comprises the following steps: chopping fresh meat, adding distilled water, uniformly stirring, adjusting the pH value, adding an enzyme for enzymolysis, deactivating the enzyme, and centrifuging to obtain supernate; filtering the obtained supernate by adopting an ultramicro filter membrane to obtain a fresh meat polypeptide primary liquid; adsorbing and eluting the obtained fresh meat polypeptide primary solution to obtain a fresh meat polypeptide concentrated solution; according to the method, the purification process is stable through real-time monitoring, regulation and control of the ultramicro filter membrane filtration process, abnormal conditions caused by fluctuation and the like possibly occurring in the preparation and purification process of the fresh meat polypeptide can be avoided, standardized production can be achieved easily, and the method is suitable for treatment of fresh meat and large-scale production of the fresh meat polypeptide.

Description

Technical Field

[0001] The present invention relates to the technical field of polypeptide compositions, and in particular to a preparation and purification process of a fresh meat polypeptide, a polypeptide composition and applications thereof, and specifically to a preparation and purification process of a fresh meat polypeptide, a polypeptide composition and applications thereof in pet food. Background Art

[0002] Protein dynamics is the study of the transformation process of intestinal protein digestion, amino acid absorption in the small intestine, and amino acid entry into the portal circulation through intestinal epithelial cells. Protein dynamics also includes the rate (speed) of the entire digestion, absorption, and metabolism process, the digestion site of nutrients in the small intestine, and the biological availability of amino acids after ingestion. Based on protein dynamics, it can be seen that changes in protein structure will cause changes in function. Polypeptides are compounds formed by α-amino acids connected together by peptide bonds. They are intermediate products when the conformation and molecular structure of proteins change after hydrolysis. Polypeptides can be divided into dipeptides, tripeptides, tetrapeptides, etc. according to the number of amino acids, until polypeptides (usually composed of 10-100 amino acid molecules). There are also literatures that call peptides composed of 2-10 amino acids oligopeptides (small molecule peptides), peptides composed of 10-50 amino acids polypeptides, and more than 50 are called proteins. Usually, compounds formed by dehydration condensation of three or more amino acid molecules are called polypeptides. Polypeptides can be divided into biologically active polypeptides and artificially synthesized polypeptides according to their sources. According to their functions, polypeptides can be divided into: movement proteins, hormone proteins, toxin proteins, polypeptide hormones, enzyme proteins, polypeptide antibiotics, etc.

[0003] Nutrients such as protein and amino acids are indispensable components of pet food, and the digestion and absorption of these nutrients will directly affect the growth and development of pets. Hydrolyzing raw protein is equivalent to pre-digestion, which can greatly improve the digestibility of protein and amino acids for pets and promote the growth of pets. Therefore, adding peptides is an effective way to improve the quality of pet food. In addition to promoting the digestion of protein by pets, adding specific peptides that are decomposed from large polypeptide chains by hydrolysis to pet food can reduce the molecular weight, reduce the antigenicity of the original protein, and reduce pet allergic reactions; it can improve the palatability and flavor characteristics of pet food, enhance the taste of pet food, and increase the intake of pet food by dogs and cats; antioxidant peptides are active peptides that can effectively inhibit or remove excess free radicals in pets. Antioxidant peptides not only have an inhibitory effect on the peroxidation of biological macromolecules such as lipids, proteins, and DNA, but also can prevent and treat a variety of pet diseases induced by free radicals. The development of efficient and stable antioxidant peptides has always been a major difficulty in the large-scale production of pet food additives. The hydrolyzates derived from meat protein show good antioxidant capacity. If the large-scale production of active peptides from fresh meat protein can be achieved, it will help the development and utilization of functional fresh meat protein in the field of pet food.

[0004] The hydrolyzate and fermentation broth obtained by direct hydrolysis of proteins are complex mixtures composed of active polypeptides and inactive peptides of different types and molecular weights. The concentration of active polypeptides is very low. The presence of inactive peptide substances will affect the activity of polypeptides and the exertion of their biological activity. Therefore, a variety of purification methods are needed to prepare high-concentration polypeptides.

[0005] Active peptides are a hot research area in disciplines such as pet nutrition. How to achieve efficient separation and purification of peptides is one of the factors that currently limit the large-scale application of peptides in pet food. There are few studies on the large-scale preparation and purification of protein hydrolysates. At present, the purification methods of peptides include column chromatography, high performance liquid chromatography and simulated membrane adsorption. Among them, column chromatography has a long reaction time and low yield; in particular, in the purification process of existing peptides, such as the technical solution disclosed in announcement number CN 104370998B, the purification process cannot be effectively monitored to ensure the normal operation of the purification process; and abnormal conditions such as fluctuations that may occur in the purification process of peptides, especially in the face of large-scale production of industrial peptides, will seriously affect the subsequent production efficiency and quality of peptides. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present invention provides a process for preparing and purifying fresh meat polypeptides, which can effectively improve the production efficiency and quality of the polypeptides.

[0007] The purpose of the present invention can be achieved through the following technical solutions: A preparation and purification process of fresh meat polypeptide comprises the following steps: (1) Take fresh meat and chop it, add distilled water, stir evenly and adjust the pH, add enzyme to hydrolyze, inactivate the enzyme, and centrifuge to obtain the supernatant; (2) filtering the supernatant obtained in step (1) through an ultra-microfiltration membrane to obtain a fresh meat polypeptide initial solution; (3) adsorbing, eluting and concentrating the fresh meat polypeptide initial solution obtained in step (2) to obtain a fresh meat polypeptide concentrated solution; The specific process of using ultra-microfiltration membrane filtration includes the following steps: Step 1: obtaining real-time purification data of fresh meat polypeptide during the purification process; wherein the real-time purification data includes a real-time flux value; Step 2: Output a purification analysis value according to the real-time flux value of each collection area. If the purification analysis value is greater than or equal to the purification analysis threshold, a purification temporary abnormality signal is generated; Step 3: Based on the currently generated temporary abnormal purification signal, set the analysis time and obtain the purification mutation value. If the purification mutation value is less than the purification mutation threshold, generate a purification regulation signal; Step 4: Based on the purification control signal, generate corresponding purification instructions, and complete the inspection or regulation of the ultra-microfiltration membrane filter according to the purification instructions; wherein the purification instructions include purification inspection instructions and purification regulation instructions.

[0008] As a further solution of the present invention: in step 1, the ultra-microfiltration membrane is divided into equal areas to obtain a plurality of purification collection areas, and the real-time flux value of each collection area is obtained.

[0009] As a further solution of the present invention: in step 2, the purification analysis value is calculated by multiplying the flux average value by the flux difference value.

[0010] As a further solution of the present invention: the process of obtaining the total flux difference is: Obtain the real-time flux value of each collection area, and calculate the real-time flux values ​​of all collection areas to obtain the total flux value; The total flux value was calculated by difference with the standard total flux value set for the purification process, and the absolute value was taken to obtain the total flux difference.

[0011] As a further solution of the present invention: the process of obtaining the flux difference is: Obtain the real-time flux value of each collection area, calculate the average of the real-time flux values ​​of all collection areas, and obtain the average flux value; The real-time flux value of each collection area is calculated separately from the flux average value, and the absolute value is taken to obtain the flux difference of each collection area. The flux differences of all collection areas are then summed up to obtain the flux difference.

[0012] As a further solution of the present invention: In step 3, the process of obtaining the purified mutation value is: The analysis time is divided into two stages, including the first stage and the second stage; Analyze the first stage, obtain the purification analysis values ​​at adjacent moments, perform difference calculation on the purification analysis values ​​at adjacent moments, obtain the purification analysis change values ​​at adjacent moments, perform mean calculation on the purification analysis change values ​​at all adjacent moments in the first stage, and obtain the purification change value of the first stage; Analyze the second stage, obtain the purification analysis values ​​at the moment before TZ and at TZ, calculate the difference between the purification analysis values ​​at the moment before TZ and at TZ, and obtain the purification change value of the second stage; The difference between the second stage purification change value and the first stage purification change value is calculated to obtain the purification change value.

[0013] As a further solution of the present invention: in step 4, when the purification control instruction is generated, the working pressure of the ultra-microfiltration membrane filter in the first stage is obtained, marked as the real-time purification pressure value, and the real-time purification pressure values ​​at adjacent moments in the first stage are calculated by difference to obtain the purification pressure change values ​​at adjacent moments; The purification factor ratio is obtained by calculating the ratio of the purification pressure change value and the purification analysis change value at the same time in the first stage, and then the variance of all the purification factor ratios in the first stage is calculated to obtain the purification factor fluctuation ratio; If the purification factor fluctuation ratio is less than the purification factor fluctuation ratio threshold, a purification pressure influence large signal is generated; According to the generated purification pressure influence large signal, the working pressure of the ultra-microfiltration membrane filter is obtained, which is recorded as FD; By formula , calculate and obtain the control pressure FT of the ultrafiltration membrane filter; make the ultrafiltration membrane filter work according to the control pressure FT, BP is the purification analysis deviation ratio.

[0014] As a further solution of the present invention: the process of obtaining the purification analysis deviation ratio is: When the purification pressure affects the generation of a large signal, the corresponding purification analysis value is obtained and marked as an abnormal purification analysis value. The difference between the abnormal purification analysis value and the purification analysis threshold is calculated to obtain the purification analysis difference. The ratio of the purification analysis difference to the purification analysis threshold is calculated to obtain the purification analysis deviation ratio BP.

[0015] The present invention also provides a polypeptide composition, comprising a fresh meat polypeptide obtained by the above-mentioned preparation and purification process of the fresh meat polypeptide, and also comprising natural fruit and vegetable polypeptide powder and mineral element absorption-promoting peptide; the polypeptide composition is composed of the following raw materials in mass fractions: 1-5 parts of natural fruit and vegetable polypeptides, 8-30 parts of fresh meat polypeptides, and 1-3 parts of mineral element absorption-promoting peptides; the fresh meat polypeptide in the polypeptide composition is a fresh meat polypeptide obtained by freeze-drying the fresh meat polypeptide concentrate obtained by the above-mentioned preparation and purification process of the fresh meat polypeptide.

[0016] The present invention also provides an application of the polypeptide composition, which is applied in pet food.

[0017] Beneficial effects of the present invention: The present invention stabilizes the purification process by real-time monitoring and regulation of the ultra-microfiltration membrane filtration process, can avoid abnormal conditions caused by fluctuations that may occur in the preparation and purification process of fresh meat polypeptides, helps to achieve standardized production, and is suitable for fresh meat processing and large-scale production of fresh meat polypeptides. Specifically: The preparation and purification process of the fresh meat polypeptide of the present invention obtains the real-time purification data of the polypeptide composition during the purification process; outputs the purification analysis value according to the real-time flux value of each collection area, and then compares the purification analysis value with the corresponding threshold value to generate a purification temporary state signal; based on the currently generated purification temporary abnormal signal, sets the analysis time and obtains the purification mutation value; the present invention monitors the filtering and purification data of the fresh meat polypeptide in real time during the purification process, analyzes it using the flux value, obtains the purification analysis value and the purification mutation value, and respectively judges the real-time purification quality and the purification change quality, so as to facilitate the real-time monitoring and control of the preparation and purification process of the fresh meat polypeptide in time, so that the quality of the fresh meat polypeptide obtained by this purification process is higher; Furthermore, based on the purification control signal, a corresponding purification instruction is generated, and according to the purification instruction, the inspection or regulation of the ultra-microfiltration membrane filter is completed; when there is an abnormality in the preparation and purification process of the fresh meat polypeptide, the present invention analyzes and determines the influencing factors, and then timely analyzes the specific regulation values ​​according to the obtained influencing factors to complete the regulation of the purification process. The purification process through real-time monitoring and regulation can effectively improve the production efficiency and quality of the polypeptide.

[0018] Based on protein dynamics, changes in protein structure will cause changes in function; unlike protein raw materials, the nitrogen source in the polypeptide composition of the present invention exists in pet food in the form of small peptides, which has a higher deposition efficiency than pet food protein composed of amino acids or complete proteins; in the polypeptide composition of the present invention, the fresh meat polypeptide has excellent antioxidant activity, and can be added to pet food as a nutritional supplement, as well as eliminate free radicals and inhibit lipid peroxidation. Moreover, the fresh meat polypeptide prepared by the preparation and purification process of the present application has the advantages of high purity and less odor, which can avoid abnormal conditions caused by local damage to the membrane and other possible fluctuations during the preparation and purification of the fresh meat polypeptide, and is suitable for large-scale production of fresh meat polypeptides; in the polypeptide composition of the present application, the natural fruit and vegetable polypeptides can regulate cell metabolism at the molecular level, have excellent antioxidant activity, can effectively remove free radicals harmful to pets, enhance the activity of immune cells, and combine with fresh meat polypeptides to help pets resist aging and disease; mineral element absorption promoting peptides are based on protein dynamics, and the peptides are easy to combine with mineral elements, which helps promote pets' absorption of Ca 2+ , Fe 2+ 、Zn 2+ And other mineral elements absorption. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a flowchart of the ultramicrofiltration membrane filtration of the present invention. DETAILED DESCRIPTION

[0020] The technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and specific embodiments.

[0021] Example 1 A preparation and purification process of a fresh meat polypeptide of the present invention comprises the following steps: (1) Take fresh meat and chop it, add distilled water, stir evenly and adjust the pH, add enzyme to hydrolyze, inactivate the enzyme, and centrifuge to obtain the supernatant; (2) filtering the supernatant obtained in step (1) through an ultra-microfiltration membrane to obtain a fresh meat polypeptide initial solution; (3) adsorbing and eluting the fresh meat polypeptide initial solution obtained in step (2) to obtain a fresh meat polypeptide concentrated solution; Specifically, the preparation and purification process of the fresh meat polypeptide comprises the following specific steps: chopping fresh meat, adding distilled water 3-6 times the mass of the fresh meat and stirring evenly, adjusting the pH to 8-9 and the temperature to 45-55°C, and adding alkaline protease for enzymolysis under heat preservation conditions, wherein the amount of enzyme added (E / S) is 2.0%, the enzymolysis is performed for 2-4 hours, the enzyme is inactivated, and after cooling, the supernatant is centrifuged at a speed of at least 8000r / min for at least 5 minutes to obtain a supernatant; the obtained supernatant is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 10ku to obtain a fresh meat polypeptide primary solution; the obtained fresh meat polypeptide primary solution is placed in a chromatography cabinet at 4°C, using AB-8 macroporous resin, and adsorbed and eluted under the conditions of a sample volume of 9-10BV, 60% ethanol as an eluent, and 3-4BV / h to obtain a fresh meat polypeptide concentrate; the obtained fresh meat polypeptide concentrate is concentrated by rotary evaporation to prepare a fresh meat polypeptide concentrate.

[0022] The fresh meat is preferably beef, mutton, pork, chicken or duck. In addition, fish and other commonly used meats for pet food processing can also be selected; the fresh meat is unfrozen fresh meat, or thawed frozen fresh meat.

[0023] like Figure 1 As shown, it is particularly noted that during the preparation process of the above process, the core purification process - the filtration process of the supernatant by the ultrafiltration membrane needs to be monitored to ensure the normal operation of the purification process, especially in the large-scale production of fresh meat polypeptides. Abnormal conditions such as fluctuations caused by factors such as pollution, damage to membrane fibers / membrane sheets, and long use time in the filtration process need to be handled in a timely manner, otherwise it will affect the subsequent production efficiency of fresh meat polypeptides and the quality of mass-produced fresh meat polypeptides.

[0024] In this application, the specific process of using ultra-microfiltration membrane filtration includes the following steps: Step 1: Obtaining real-time purification data of fresh meat peptides during the purification process; Wherein, the real-time purification data includes real-time flux value; In some embodiments, the ultra-microfiltration membrane is divided into multiple purification collection areas in an equal area manner, and then a flow meter is used to obtain the real-time flux value of each collection area in the filtration and purification process of the fresh meat polypeptide; Step 2: Output a purification analysis value according to the real-time flux value of each collection area, and then compare the purification analysis value with the corresponding threshold value to generate a purification temporary state signal; Wherein, the purification temporary state signal includes the purification temporary normal signal or the purification temporary abnormal signal; In some embodiments, the real-time flux value of each collection area is obtained, the total flux difference value and the flux difference value are analyzed to obtain the product of the flux average value and the flux difference value to obtain the purification analysis value; Compare the purification analysis value to the purification analysis threshold: If the purification analysis value is greater than or equal to the purification analysis threshold, it means that the fresh meat polypeptide does not meet the process standards during the purification process, and a purification temporary abnormality signal is generated; If the purification analysis value is less than the purification analysis threshold, it means that the fresh meat polypeptide meets the process standards during the purification process, and a purification temporary normal signal is generated; First, specifically, the process of obtaining the total flux difference is: Obtain the real-time flux value of each collection area, and calculate the real-time flux values ​​of all collection areas to obtain the total flux value; The total flux value is calculated by difference with the standard total flux value set in the purification process, and the absolute value is taken to obtain the total flux difference; Second, specifically, the process of obtaining the flux difference is: Obtain the real-time flux value of each collection area, calculate the average of the real-time flux values ​​of all collection areas, and obtain the average flux value; The real-time flux value of each collection area is calculated by difference with the average flux value, and the absolute value is taken to obtain the flux difference of each collection area. Then the flux difference values ​​of all collection areas are summed up to obtain the flux difference value. It should be explained that the total flux difference indicates the difference between the state of filtration purification and the standard state during the ultrafiltration membrane purification of the fresh meat polypeptide. The larger the total flux difference, the worse the polypeptide purification effect, and the lower the quality of the obtained fresh meat polypeptide. The flux difference indicates the degree of difference in filtration and purification between the various collection areas during the ultrafiltration membrane purification process of the current fresh meat polypeptide. The larger the flux difference, the worse the polypeptide purification effect, and thus the lower the quality of the obtained fresh meat polypeptide. Step 3: Based on the currently generated temporary abnormal purification signal, set the analysis time, obtain the purification mutation value, and then compare the purification mutation value with the corresponding threshold to generate a purification control signal; Wherein, the purification control signal includes a purification pause signal or a purification regulation signal; In some embodiments, when a temporary abnormal signal of purification is obtained, an analysis time is set (the interval of the analysis time is [T1, T n ], where T1 is the starting point of the analysis time, T n is the end point of the analysis time, T n That is, the time when the current purification temporary abnormal signal is generated, and the length of the interval is set by the technician), obtain the purification analysis value within the analysis time, and obtain the purification mutation value after analysis; The obtained purified mutation value is compared with the purified mutation threshold; If the purification mutation value is greater than or equal to the purification mutation threshold, a purification pause signal is generated; If the purification mutation value is less than the purification mutation threshold, a purification regulation signal is generated; Specifically, the process of obtaining the purified mutation value is as follows: The analysis time is divided into two stages, including the first stage and the second stage. Specifically, the time interval of the first stage is [T1,T n-1 ], the time interval of the second stage is [T n-1 ,T n ]; Analyze the first stage, obtain the purification analysis values ​​at adjacent moments, perform difference calculation on the purification analysis values ​​at adjacent moments, obtain the purification analysis change values ​​at adjacent moments, perform mean calculation on the purification analysis change values ​​at all adjacent moments in the first stage, and obtain the purification change value of the first stage; Analyze the second stage, obtain the purification analysis values ​​at the moment before TZ and at TZ, calculate the difference between the purification analysis values ​​at the moment before TZ and at TZ, and obtain the purification change value of the second stage; The difference between the second stage purification change value and the first stage purification change value is calculated to obtain a purification change value; It needs to be explained that the purification mutation value represents the difference between the change rate of the purification analysis value at the current moment and the change rate of the purification analysis value in the previous time. If the purification mutation value is larger, it means that during the purification of fresh meat polypeptide by ultrafiltration membrane, the filtration speed of the polypeptide solution deviates greatly from the preset standard, which is likely to be caused by leakage such as damage to the ultrafiltration membrane; on the contrary, if the purification mutation value is smaller, when a temporary abnormal purification signal appears, it is not caused by leakage such as damage to the ultrafiltration membrane; The technical scheme of the above purification process is as follows: obtaining real-time purification data of fresh meat polypeptides during the purification process; outputting a purification analysis value according to the real-time flux value of each collection area, and then comparing the purification analysis value with the corresponding threshold value to generate a purification temporary state signal; setting the analysis time based on the currently generated purification temporary abnormality signal to obtain the purification mutation value; the present invention monitors the filtering and purification data of the fresh meat polypeptides in real time during the purification process, and uses the flux value for analysis to obtain the purification analysis value and the purification mutation value, and respectively judges the real-time purification quality and the purification change quality, so as to facilitate the real-time monitoring and control of the preparation and purification process of the fresh meat polypeptides in a timely manner, so that the quality of the fresh meat polypeptides obtained through such purification is higher; Step 4: Generate corresponding purification instructions based on the purification control signal, and complete the inspection or regulation of the ultra-microfiltration membrane filter according to the purification instructions; Wherein, purification instructions include purification inspection instructions and purification control instructions; In some embodiments, when a purification pause signal is obtained, the ultra-microfiltration membrane filter generates a purification inspection instruction accordingly; when the purification inspection instruction is obtained, the ultra-microfiltration membrane of the ultra-microfiltration membrane filter is inspected first, and if damage occurs, the ultra-microfiltration membrane is replaced; When a purification control signal is obtained, the ultra-microfiltration membrane filter generates a corresponding purification control instruction; when the purification control instruction is obtained, the working pressure of the ultra-microfiltration membrane filter is obtained, and its influencing factors are judged. If there is an influence, the working pressure of the ultra-microfiltration membrane filter is regulated; Specifically, when the purification control instruction is generated, the working pressure of the ultra-microfiltration membrane filter in the first stage is obtained, marked as the real-time purification pressure value, and the real-time purification pressure values ​​at adjacent moments in the first stage are calculated by difference to obtain the purification pressure change values ​​at adjacent moments; The purification factor ratio is obtained by calculating the ratio of the purification pressure change value and the purification analysis change value at the same time in the first stage, and then the variance of all the purification factor ratios in the first stage is calculated to obtain the purification factor fluctuation ratio; comparing the purification factor fluctuation ratio with a purification factor fluctuation ratio threshold; If the purification factor fluctuation ratio is greater than or equal to the purification factor fluctuation ratio threshold, it means that in the first stage, the purification factor ratio at each time has a large fluctuation, that is, the influence between the purification pressure and the purification state change is small, and a small purification pressure influence signal is generated; If the purification factor fluctuation ratio is less than the purification factor fluctuation ratio threshold, it means that in the first stage, the purification factor ratio at each time has a small fluctuation, that is, the influence between the purification pressure and the purification state change is large, and a purification pressure influence signal is generated; According to the generated purification pressure influence large signal, the working pressure of the ultra-microfiltration membrane filter is obtained, which is recorded as FD; And, obtaining the purification analysis value corresponding to the generation of the large signal affected by the purification pressure, marking it as an abnormal purification analysis value, performing a difference calculation between the abnormal purification analysis value and the purification analysis threshold to obtain a purification analysis difference, and then performing a ratio calculation between the purification analysis difference and the purification analysis threshold to obtain a purification analysis deviation ratio BP; By formula , calculate and obtain the regulating pressure FT of the ultra-microfiltration membrane filter; make the ultra-microfiltration membrane filter work according to the regulating pressure FT; The technical scheme of the above-mentioned purification process generates corresponding purification instructions based on the purification control signal, and completes the inspection or regulation of the ultramicrofiltration membrane filter according to the purification instructions; in the case of abnormal conditions in the preparation and purification process of fresh meat polypeptides, the present invention analyzes and determines the influencing factors, and then timely analyzes the specific regulation values ​​according to the obtained influencing factors to complete the regulation of the purification process, thereby ensuring that the fresh meat polypeptides obtained through the purification process monitored and regulated in real time have high quality.

[0025] Example 2 A polypeptide composition, comprising fresh meat polypeptide powder obtained by the above-mentioned preparation and purification process of fresh meat polypeptide, as well as natural fruit and vegetable polypeptide powder and mineral element absorption promoting peptide powder; specifically, the polypeptide composition is composed of the following raw materials in mass fractions: 3 parts of natural fruit and vegetable polypeptide, 20 parts of fresh meat polypeptide and 2 parts of mineral element absorption promoting peptide.

[0026] Among them, natural fruit and vegetable peptides can regulate cell metabolism at the molecular level, have excellent antioxidant activity, effectively remove free radicals that are harmful to pets, enhance the activity of immune cells, and help pets resist aging and diseases; the preparation method thereof can refer to the prior art such as CN 109673751 A; Fresh meat peptides have excellent antioxidant activity and can be added to pet food as a nutritional supplement, as well as to eliminate free radicals and inhibit lipid peroxidation; Mineral element absorption peptide is based on protein dynamics. The peptide is easy to combine with mineral elements to promote pets' absorption of Ca 2+ , Fe 2+ 、Zn 2+ The absorption of mineral elements such as calcium and phosphorylation is promoted by the mineral element absorption peptide, which is easy to bind to mineral elements. Since pets have a relatively large demand for mineral elements, protein in their daily diet cannot provide an effective function of promoting the absorption of mineral elements. The present invention uses α-lactalbumin with metal ion binding properties and whey protein containing α-lactalbumin as a raw material to prepare a mineral element absorption-promoting peptide that is easy to bind to mineral elements. The central position of the mineral element absorption-promoting peptide contains a phosphorylated serine group and a glutamyl residue, which can bind to Ca at the lower end of the small intestine of pets near pH 7-8. 2+ , Fe 2+ 、Zn 2+It combines with metal ions such as calcium, iron, and zinc to prevent them from precipitating with phosphate, thereby increasing the concentration of soluble calcium, iron, and zinc in the small intestine of the pet, and promoting the absorption and utilization of these essential metal ions.

[0027] The preparation method of the mineral element absorption promoting peptide is as follows: weigh a certain amount of whey protein, prepare a solution with distilled water at a substrate concentration of 7.0% (w / v), adjust the pH to 8.0 under a constant temperature condition of 45°C, add trypsin at a substrate to enzyme mass ratio of 70:1 for hydrolysis, and hydrolyze at a constant temperature for 100 minutes. During the hydrolysis process, adjust the pH every 10 minutes to maintain the pH at 7.5-8.5. After the hydrolysis is completed, kill the enzyme, centrifuge to obtain the supernatant, transfer the supernatant obtained by centrifugation to a 10ku ultrafiltration tube, centrifuge at 4500g and 4°C for 30 minutes, and ultrafilter and freeze-dry to obtain the mineral element absorption promoting peptide.

[0028] Example 3 The difference from the above-mentioned Example 2 is that the polypeptide composition is composed of the following raw materials in mass fractions: 1 part of natural fruit and vegetable polypeptides, 8 parts of fresh meat polypeptides, and 1 part of mineral element absorption-promoting peptides.

[0029] Example 4 The difference from the above-mentioned Example 2 is that the polypeptide composition is composed of the following raw materials in mass fractions: 5 parts of natural fruit and vegetable polypeptides, 30 parts of fresh meat polypeptides, and 3 parts of mineral element absorption-promoting peptides.

[0030] Example 5 An application of a polypeptide composition, wherein the polypeptide composition is applied to pet food, such as directly applied to pet wet food, or applied to the preparation of pet dry food and pet snacks.

[0031] The embodiments of the present invention are described in detail above, but the contents described are only preferred embodiments of the present invention and cannot be considered to limit the scope of implementation of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the patent coverage of the present invention.

Claims

1. A process for preparing and purifying fresh meat polypeptides, characterized in that: The following steps are involved: (1) Take fresh meat and chop it, add distilled water, stir evenly and adjust the pH, add enzyme to hydrolyze, inactivate the enzyme, and centrifuge to obtain the supernatant; (2) filtering the supernatant obtained in step (1) through an ultramicrofiltration membrane to obtain a fresh meat polypeptide initial solution; (3) adsorbing, eluting, and concentrating the fresh meat polypeptide initial solution obtained in step (2) to obtain a fresh meat polypeptide concentrated solution; The specific process of using ultra-microfiltration membrane filtration includes the following steps: Step 1: obtaining real-time purification data of fresh meat polypeptide during the purification process; wherein the real-time purification data includes real-time flux value; Step 2: Output the purification analysis value based on the real-time flux value of each collection area. If the purification analysis value is greater than or equal to the purification analysis threshold, a purification temporary abnormality signal is generated; Step 3: Based on the currently generated purification temporary abnormality signal, set the analysis time and obtain the purification mutation value. If the purification mutation value is less than the purification mutation threshold, generate a purification regulation signal; Step 4: Based on the purification control signal, generate corresponding purification instructions, and complete the inspection or regulation of the ultrafiltration membrane filter according to the purification instructions; wherein the purification instructions include purification inspection instructions and purification regulation instructions.

2. The process for preparing and purifying fresh meat polypeptide according to claim 1, characterized in that: In step 1, the ultrafiltration membrane is divided into equal areas to obtain multiple purification collection areas, and the real-time flux value of each collection area is obtained.

3. The process for preparing and purifying fresh meat polypeptide according to claim 1, characterized in that: In step 2, the purification analysis value is calculated by multiplying the flux mean value by the flux difference value.

4. The process for preparing and purifying fresh meat polypeptide according to claim 3, characterized in that: The process of obtaining the total flux difference is: Obtain the real-time flux value of each collection area, and calculate the sum of the real-time flux values ​​of all collection areas to obtain the total flux value; The total flux value was calculated by difference with the standard total flux value set in the purification process, and the absolute value was taken to obtain the total flux difference.

5. The process for preparing and purifying fresh meat polypeptide according to claim 4, characterized in that: The process of obtaining the flux difference is: Obtain the real-time flux value of each collection area, calculate the average of the real-time flux values ​​of all collection areas, and obtain the average flux value; The real-time flux value of each collection area is calculated separately from the flux average value, and the absolute value is taken to obtain the flux difference of each collection area. The flux differences of all collection areas are then summed up to obtain the flux difference.

6. The process for preparing and purifying fresh meat polypeptide according to claim 1, characterized in that: In step 3, the process of obtaining the purified mutation value is as follows: The analysis time is divided into two stages, including the first stage and the second stage. The time interval of the first stage is [T1,T n-1 ], the time interval of the second stage is [T n-1 ,T n ]; Analyze the first stage to obtain purification analysis values ​​at adjacent moments, perform difference calculation on the purification analysis values ​​at adjacent moments to obtain purification analysis change values ​​at adjacent moments, and perform mean calculation on the purification analysis change values ​​at all adjacent moments in the first stage to obtain the purification change value for the first stage; Analyze the second stage, obtain the purification analysis values ​​at the moment before TZ and at TZ, calculate the difference between the purification analysis values ​​at the moment before TZ and at TZ, and obtain the purification change value of the second stage; The difference between the second stage purification change value and the first stage purification change value is calculated to obtain the purification change value.

7. The process for preparing and purifying fresh meat polypeptide according to claim 1, characterized in that: In step 4, when the purification control instruction is generated, the working pressure of the ultrafiltration membrane filter in the first stage is obtained and marked as the real-time purification pressure value. The real-time purification pressure values ​​at adjacent moments in the first stage are differentially calculated to obtain the purification pressure change values ​​at adjacent moments. The purification factor ratio is obtained by calculating the ratio of the purification pressure change value to the purification analysis change value at the same time in the first stage, and the purification factor fluctuation ratio is obtained by calculating the variance of all the purification factor ratios in the first stage; If the purification factor fluctuation ratio is less than the purification factor fluctuation ratio threshold, a purification pressure influence large signal is generated; According to the generated purification pressure influence signal, the working pressure of the ultrafiltration membrane filter is obtained and recorded as FD; By formula , calculate and obtain the control pressure FT of the ultrafiltration membrane filter; make the ultrafiltration membrane filter work according to the control pressure FT, BP is the purification analysis deviation ratio.

8. The process for preparing and purifying fresh meat polypeptide according to claim 7, characterized in that: The process of obtaining the deviation ratio of purification analysis is as follows: When the purification pressure affects the generation of a large signal, the corresponding purification analysis value is obtained and marked as an abnormal purification analysis value. The difference between the abnormal purification analysis value and the purification analysis threshold is calculated to obtain the purification analysis difference. The ratio of the purification analysis difference to the purification analysis threshold is then calculated to obtain the purification analysis deviation ratio BP.

9. A polypeptide composition, characterized in that: The fresh meat polypeptide is obtained by the preparation and purification process of the fresh meat polypeptide according to any one of claims 1 to 8, and further comprises natural fruit and vegetable polypeptides and mineral element absorption-promoting peptides; the polypeptide composition is composed of the following raw materials in mass fractions: 1-5 parts of natural fruit and vegetable polypeptides, 8-30 parts of fresh meat polypeptides, and 1-3 parts of mineral element absorption-promoting peptides.

10. An application of a polypeptide composition, characterized in that: The polypeptide composition is used in pet food.

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