Low-fat, low-cost, high-purity and high-quality elastin peptide and preparation method thereof

Abstract

The invention belongs to the technical field of active substance extraction, and particularly relates to low-fat, low-cost, high-purity and high-quality elastin peptide and a preparation method thereof. The alkali liquor and the sodium carbonate solution are mixed and matched for use, the overall concentration is low, the degreasing agent is more environmentally friendly, and the degreasing effect is better by using the microwave heating technology. The protease is immobilized by using the microspheres, and the enzyme preparation can be fully recycled and repeatedly used, so that the production cost is greatly reduced. The elastin enzymatic hydrolysate is separated, purified and refined by using an ion exchange resin tandem chromatography technology, other impurities are efficiently removed, and the prepared elastin peptide is relatively high in purity. The high-voltage pulsed electric field technology (PEF) is used for sterilization, the sterilization effect is better, the influence of a traditional heat sterilization mode on the color, luster and taste of the product is avoided, and the quality of elastin peptide is better maintained.

Description

Technical Field

[0001] The invention belongs to the technical field of active substance extraction, and specifically relates to a low-fat, low-cost, high-purity and high-quality elastin peptide and a preparation method thereof. Background Art

[0002] Elastin is an important extracellular matrix protein and the main component of elastic fibers. It is present in large quantities in tissues and organs that are often deformed by force, such as the lungs, aorta, skin, ligaments, and arterial bulbs of mammals. Elastin contains a special cross-linking structure, desmosin and isodesmosin. The soluble precursor protein tropoelastin of elastin is cross-linked by desmosin and isodesmosin to form a dense protein structure. Both proteins are water-insoluble proteins. Due to their dense structure and hydrophobicity, they have good ductility and elasticity, and can resist hydrolysis by most hydrolases. As amino acids unique to elastin, the content of desmosin and isodesmosin has an important influence on the biosynthesis and physiological function of elastin.

[0003] Elastin tissue is often rich in lipid substances, which seriously affects the use effect of elastin. The enzymatic hydrolysis efficiency and purity of elastin are improved by degreasing. At present, the raw materials for preparing elastin peptides are basically pretreated with alkaline solution, but the concentration used is relatively high. Chinese patent application CN106519020A uses 0.5% sodium hydroxide solution to treat raw materials containing elastin at 85-95°C, and Chinese patent CN108977488B uses 3-6% sodium carbonate solution to treat fresh bovine neck ligament to remove fat and connective tissue, but the waste liquid generated by the above method puts great pressure on the environment. In addition, the current preparation of elastin peptides directly uses free protease liquid or powder for enzymatic hydrolysis. The enzyme preparation has poor ability to resist adverse external environment, easily causes loss of activity and is not easy to recycle and reuse, and the production cost is relatively high. Moreover, the conventional method does not carry out the purification and refining process, and the content of desmosin and isodesmosin in the prepared elastin peptide is low. The traditional heat sterilization method is easy to cause glycosylation reaction in the feed liquid, affecting the color and taste of the product, resulting in a decrease in product quality.

[0004] Therefore, how to further reduce the fat content of elastin peptide products and reduce costs while retaining the activity of elastin peptides during the preparation of elastin peptides is an urgent problem to be solved in the art. Summary of the invention

[0005] The present invention aims at the defects and shortcomings in the prior art and provides a low-fat, low-cost, high-purity and high-quality elastin peptide and a preparation method thereof.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] In a first aspect, the present invention provides a method for preparing a low-fat, low-cost, high-purity and high-quality elastin peptide, the preparation method comprising the following steps:

[0008] (1) Pretreatment: Cut the cleaned elastin-rich animal tissue into small pieces, add 8-12 times the weight of an alkaline solution, soak at room temperature for 2-6 hours, and wash with purified water until the conductivity of the effluent is within 1000 μs / cm, thereby obtaining a pretreated elastin tissue, wherein the alkaline solution is a mixed solution of alkali and sodium carbonate, and the mass concentration ratio of alkali to sodium carbonate is (0.05-0.1%): (0.5-1%);

[0009] (2) Heating aging: After cleaning, the pretreated elastin tissue prepared in step (1) is added with 1-3 times the weight of purified water, and then the material is heated by microwave heating. The microwave power is constant at 500-800 W. The microwave treatment is performed in an intermittent manner. After working for 2-4 minutes, it is rested for 2-4 minutes. One microwave action and rest is considered as one cycle. 6-8 cycles are performed.

[0010] (3) Two-phase separation: The liquid after heat extraction is allowed to stand and the two phases are separated, with the oil phase in the upper layer and the water phase in the lower layer. The oil phase is fully removed by liquid separation;

[0011] (4) Enzymatic hydrolysis: adding 5-15% of the weight of the pretreated elastin tissue to the aqueous phase, adjusting the temperature to 50-65° C., and performing enzymatic hydrolysis for 3-8 hours to obtain an enzymatic hydrolyzate containing elastin peptides;

[0012] (5) solid-liquid separation: the enzymatic hydrolyzate containing elastin peptides prepared in step (4) is subjected to solid-liquid separation using a 50-100 nm ceramic membrane to obtain a clear and transparent enzymatic hydrolyzate solution;

[0013] (6) Decolorization and purification: The enzymatic hydrolyzate prepared in step (5) is sequentially passed through a series of chromatography columns filled with a cation exchange resin, anion exchange resin, and cation exchange resin, with a flow rate controlled at 4-6 BV / h, to remove other impurity protein peptides including collagen peptides, and to prepare an elastin peptide solution, wherein the cation exchange resin, anion exchange resin, and cation exchange resin are respectively weak acid type, strong base type, and weak acid type ion exchange resins, and the column loading volume ratio is 1:1.2:1-1:1.5:1;

[0014] (7) Sterilization: sterilize the elastin peptide solution prepared in step (6) using high voltage pulsed electric field technology (PEF), with an electric field strength of 15-30 KV / cm and a pulse number of 100-350;

[0015] (8) Drying: spray drying the sterilized elastin peptide solution prepared in step (7) to obtain elastin peptide powder.

[0016] As an optional mode, in the above preparation method, in step (1), the elastin-rich animal tissue is the aorta, skin, ligament of mammals such as cows or pigs, or the bulbus arteriosus of the heart of fish.

[0017] Preferably, the elastin-rich animal tissue is fish heart bulb.

[0018] Preferably, the elastin-rich animal tissue is cut into blocks of (2-4) mm×(2-4) mm×(2-4) mm.

[0019] As an optional manner, in the above preparation method, in step (1), the base is sodium hydroxide or potassium hydroxide.

[0020] Preferably, the base is sodium hydroxide.

[0021] As an optional mode, in the above preparation method, in step (4), the immobilized protease is selected from one or more of the following: alkaline protease, papain, bromelain, neutral protease, flavor protease, trypsin or pepsin.

[0022] As an optional mode, in the above preparation method, the immobilized protease is selected from alkaline protease and papain.

[0023] As an optional manner, in the above preparation method, the preparation method of the immobilized protease in step (4) comprises the following steps: based on the total mass of the immobilized protease, the mass concentration of sodium alginate is 2-4%, the mass concentration of chitosan is 3-5%, the mass concentration of calcium chloride is 2-4%, the mass concentration of glutaraldehyde is 0.5-1%, the mass concentration of protease is 0.5-0.8%, the cross-linking time is 1-4h, and the immobilization temperature is 35-45°C, so as to prepare the microsphere immobilized protease.

[0024] The thermodynamic stability and enzymatic properties of the microsphere-immobilized protease prepared by the invention are relatively good, and the enzyme activity can still be maintained above 65% after being repeatedly used for 5 times.

[0025] As an optional method, in the above preparation method, the method for recovering the immobilized protease in step (4) comprises the following steps: passing the enzymatic hydrolysate containing the immobilized protease through a 40-60 mesh standard sieve to recover the microsphere immobilized protease, rinsing with pure water and refrigerating for repeated use.

[0026] In a second aspect, the present invention provides an elastin peptide prepared by the preparation method described in the first aspect.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] (1) The present invention uses a mixture of alkali solution and sodium carbonate solution, which has a low overall concentration and is more environmentally friendly. The degreasing effect is better using microwave heating technology.

[0029] (2) The present invention uses microspheres to immobilize proteases, and the enzyme preparation can be fully recovered and reused multiple times, greatly reducing production costs.

[0030] (3) The ion exchange resin series chromatography technology is used to separate, purify and refine the elastase hydrolysate, effectively removing other impurities, and the prepared elastin peptide has a high purity.

[0031] (4) The present invention uses high-voltage pulsed electric field technology (PEF) for sterilization, which has a better sterilization effect, avoids the influence of traditional thermal sterilization methods on product color and taste, and better maintains the quality of elastin peptides. DETAILED DESCRIPTION

[0032] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0033] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0034] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0035] Preparation Example:

[0036] Example 1

[0037] A method for preparing low-fat, low-cost, high-purity and high-quality elastin peptides, comprising the following steps:

[0038] (1) Pretreatment: adding 8 times the weight of an alkaline solution to the cleaned tilapia heart arterial bulb, soaking it at room temperature for 4 hours, and washing it with purified water until the conductivity of the effluent is within 1000 μs / cm, thereby obtaining the pretreated tilapia heart arterial bulb, wherein the alkaline solution is a mixed solution of sodium hydroxide and sodium carbonate, and the mass concentration ratio of sodium hydroxide to sodium carbonate is 0.05%:1%;

[0039] (2) Heating aging: Add 2 times the weight of purified water to the cleaned tilapia heart artery bulb, and then use microwave heating to heat the material. The microwave power is constant at 500 W. The microwave treatment is intermittent. After working for 4 minutes, it is rested for 4 minutes. One cycle is composed of one microwave action and one rest. Six cycles are performed.

[0040] (3) Two-phase separation: The liquid after heat extraction is allowed to stand and the two phases are separated, with the oil phase in the upper layer and the water phase in the lower layer. The oil phase is fully removed by liquid separation;

[0041] (4) Enzymatic hydrolysis: adding 5% of the weight of the pretreated tilapia cardiac arterial bulb microsphere-immobilized alkaline protease and 10% of the weight of the pretreated tilapia cardiac arterial bulb microsphere-immobilized papain to the aqueous phase, adjusting the temperature to 55° C., and performing enzymatic hydrolysis for 5 h to obtain an enzymatic hydrolyzate containing elastin peptides;

[0042] (5) solid-liquid separation: the enzymatic hydrolyzate containing elastin peptides prepared in step (4) is subjected to solid-liquid separation using a ceramic membrane (100 nm) to prepare a clear and transparent enzymatic hydrolyzate solution;

[0043] (6) Decolorization and purification: The enzymatic hydrolyzate prepared in step (5) is sequentially passed through a series of chromatography columns filled with cation exchange resin-anion exchange resin-cation exchange resin, with the flow rate controlled at 4 BV / h, to remove other impurity protein peptides including collagen peptides, and to prepare an elastin peptide solution, wherein the cation exchange resin-anion exchange resin-cation exchange resin are respectively weak acid-strong base-weak acid ion exchange resins, and the column loading volume ratio is 1:1.2:1;

[0044] (7) Sterilization: The elastin peptide solution prepared in step (6) is sterilized using a high voltage pulse electric field technique (PEF) with an electric field strength of 20 kV / cm and a pulse number of 200;

[0045] (8) Drying: spray drying the sterilized elastin peptide solution prepared in step (7) to obtain elastin peptide powder.

[0046] The preparation method of the immobilized protease in step (4) comprises the following steps: based on the total mass of the immobilized protease, the mass concentration of sodium alginate is 3%, the mass concentration of chitosan is 3%, the mass concentration of calcium chloride is 3.5%, the mass concentration of glutaraldehyde is 1%, the mass concentration of protease is 0.6%, the cross-linking time is 1h, and the immobilization temperature is 40°C, so as to prepare the microsphere immobilized protease. The thermodynamic stability and enzymatic properties of the prepared microsphere immobilized protease are relatively good, and the enzyme activity can still be maintained at more than 65% after being reused 5 times.

[0047] The method for recovering the immobilized protease in step (4) comprises the following steps: passing the enzymatic solution containing the immobilized protease through a 40-mesh standard sieve to recover the microsphere immobilized protease, and rinsing with pure water and storing in a refrigerator for repeated use.

[0048] Example 2

[0049] A method for preparing low-fat, low-cost, high-purity and high-quality elastin peptides, comprising the following steps:

[0050] (1) Pretreatment: adding 8 times the weight of alkaline solution to the cleaned tilapia heart arterial bulb and soaking it at room temperature for 4 hours, and washing it with purified water until the conductivity of the effluent is within 1000 μs / cm, thereby pretreating the tilapia heart arterial bulb, wherein the alkaline solution is a mixed solution of sodium hydroxide and sodium carbonate, and the mass concentration ratio of sodium hydroxide to sodium carbonate is 0.1%:0.5%;

[0051] (2) Heating aging: Add 2 times the weight of purified water to the cleaned tilapia heart artery bulb, and then use microwave heating to heat the material. The microwave power is constant at 800 W. The microwave treatment is intermittent. After working for 2 minutes, it is rested for 2 minutes. The execution of one microwave action and rest is one cycle, and 6 cycles are performed;

[0052] (3) Two-phase separation: The liquid after heat extraction is allowed to stand and the two phases are separated, with the oil phase in the upper layer and the water phase in the lower layer. The oil phase is fully removed by liquid separation;

[0053] (4) Enzymatic hydrolysis: adding 5% of the weight of the pretreated tilapia cardiac arterial bulb microsphere-immobilized alkaline protease and 10% of the weight of the pretreated tilapia cardiac arterial bulb microsphere-immobilized papain to the aqueous phase, adjusting the temperature to 55° C., and performing enzymatic hydrolysis for 5 h to obtain an enzymatic hydrolyzate containing elastin peptides;

[0054] (5) solid-liquid separation: the enzymatic hydrolyzate containing elastin peptides prepared in step (4) is subjected to solid-liquid separation using a ceramic membrane (100 nm) to prepare a clear and transparent enzymatic hydrolyzate solution;

[0055] (6) Decolorization and purification: The enzymatic hydrolyzate prepared in step (5) is sequentially passed through a series of chromatography columns filled with cation exchange resin-anion exchange resin-cation exchange resin, with the flow rate controlled at 4 BV / h, to remove other impurity protein peptides including collagen peptides, and to prepare an elastin peptide solution, wherein the cation exchange resin-anion exchange resin-cation exchange resin are respectively weak acid-strong base-weak acid ion exchange resins, and the column loading volume ratio is 1:1.2:1;

[0056] (7) Sterilization: The elastin peptide solution prepared in step (6) is sterilized using a high voltage pulse electric field technique (PEF) with an electric field strength of 30 kV / cm and a pulse number of 100;

[0057] (8) Drying: spray drying the sterilized elastin peptide solution prepared in step (7) to obtain elastin peptide powder.

[0058] The preparation method of the immobilized protease in step (4) comprises the following steps: based on the total mass of the immobilized protease, the mass concentration of sodium alginate is 3%, the mass concentration of chitosan is 3%, the mass concentration of calcium chloride is 3.5%, the mass concentration of glutaraldehyde is 1%, the mass concentration of protease is 0.6%, the cross-linking time is 1h, and the immobilization temperature is 40°C, so as to prepare the microsphere immobilized protease. The thermodynamic stability and enzymatic properties of the prepared microsphere immobilized protease are relatively good, and the enzyme activity can still be maintained at more than 65% after being reused 5 times.

[0059] The method for recovering the immobilized protease in step (4) comprises the following steps: passing the enzymatic solution containing the immobilized protease through a 40-mesh standard sieve to recover the microsphere immobilized protease, and rinsing with pure water and storing in a refrigerator for repeated use.

[0060] Comparative Example 1

[0061] The step (2) in Example 1 was changed to heat aging: 2 times the weight of purified water was added to the cleaned tilapia heart arterial bulb and stirred at 90° C. for 4 h.

[0062] Comparative Example 2

[0063] The step (4) in Example 1 was changed to enzymatic hydrolysis: 0.5% alkaline protease by weight of tilapia heart arterial bulb and 1.0% papain by weight of tilapia heart arterial bulb were added to the aqueous phase, the temperature was adjusted to 55° C., and the enzymatic hydrolysis was performed for 5 hours to obtain an enzymatic hydrolyzate containing elastin peptides.

[0064] Step (5) was changed to enzyme inactivation: the temperature of the enzymatic hydrolysate was raised to 90°C and kept warm for 15 min.

[0065] Comparative Example 3

[0066] The step (6) in Example 1 is changed to decolorization and nanofiltration: the clarified enzymatic hydrolysate is decolorized by an activated carbon fiber membrane and desalted by a nanofiltration membrane with a molecular weight of 200D.

[0067] Comparative Example 4

[0068] The step (7) in Example 1 was changed to sterilization: the enzymatic hydrolysate was treated at 121° C. for 5 s using an ultra-high temperature instantaneous sterilization (UHT) device.

[0069] Effect embodiment:

[0070] Effect Example 1

[0071] The test indicators and their detection and analysis methods involved in each embodiment and comparative example prepared in the above preparation example of the present invention are as follows:

[0072] The present invention uses the second method of fat determination in food of GB5009.6-2016 to determine the fat content in elastin peptide.

[0073] The content of desmosin / isodesmosin was determined by referring to the method in Appendix A of T / CHC 1010-2023 elastin peptide. The specific method is as follows: the sample was hydrolyzed under acidic conditions and determined using high performance liquid chromatography-mass spectrometry (HPLC-MS / MS). The retention time and mass-to-charge ratio of desmosin and isodesmosin standards were used for qualitative and confirmation, and the external standard method was used for quantitative analysis. Chromatographic conditions: Atlantis dC18 (3μm, 150mm×2.1mm), column temperature 30℃, flow rate 0.3mL / min, mobile phase A: aqueous solution (containing 7mM heptafluorobutyric anhydride, 5mM ammonium acetate), mobile phase B: 80% acetonitrile (containing 7mM heptafluorobutyric anhydride, 5mM ammonium acetate); elution program: 0-14min mobile phase A gradient elution from 100% to 30%, 14-22min elution to 100% mobile phase A. Mass spectrometry conditions: electrospray ion source, positive ion mode, ion source spray voltage 3.5 kV, capillary temperature 320 °C, collision energy 35 eV, desmosine parent ion (m / z) 526.3, daughter ion (m / z) 481.3, isodesmosine parent ion (m / z) 526.3, daughter ion

[0074] (m / z)397.3.

[0075] Desmosin content (%) = desmosin mass in sample / sample mass × 100.

[0076] Isodesmosin content (%) = mass of isodesmosin in sample / mass of sample × 100.

[0077] The total colony count is determined according to GB 4789.2.

[0078] The whiteness was measured using a JC-BD1A benchtop whiteness meter.

[0079] The determination of ash content shall refer to GB 5009.4-2016.

[0080] The results of the fat content percentage, desmosin content %, isodesmosin content %, total bacterial count, whiteness and ash content % in the samples prepared in Examples 1-2 and Comparative Examples 1-4 are compared in Table 1.

[0081] Table 1 Physical and chemical indicators of samples prepared in various embodiments and comparative examples

[0082]

[0083] As can be seen from Table 1, the fat content of the samples prepared in the embodiment is relatively low, all less than 0.5%, and the fat content in Comparative Example 1 is high, indicating that microwave heating can better interrupt the natural combination between the fat component and the protein component in the heart arterial bulb, and remove grease more efficiently. From the data of the embodiment and comparative example 2, it can be seen that the same amount of enzyme preparation added, the experimental effect of the immobilized enzyme and the free protease preparation is consistent, and the immobilized enzyme can be recycled and reused many times. In addition, the process of heating and killing enzymes is omitted, which greatly reduces production time and cost. From the data of the embodiment and comparative example 3, it can be seen that the content of desmosin / isodesmosin in the sample in comparative example 3 is significantly reduced, and the whiteness is significantly lower than that in the embodiment, indicating that the ion exchange resin can more effectively adsorb pigments, separate impurities, reduce ash content, and effectively improve the purity of elastin peptides. From the data of the embodiment and comparative example 4, it can be seen that the color of the elastin peptide sterilized at high temperature is darker, the whiteness is significantly reduced, and the high temperature sterilization may damage the elastin peptide, and the content of desmosin and isodesmosin is reduced. Compared with traditional high temperature sterilization, high voltage pulse electric field technology (PEF) has better sterilization effect and higher product whiteness.

[0084] Effect Example 2

[0085] Test method: Recruitment age is between 35-45 years old, BMI is between 18.5-27.9 kg / m 2 A total of 120 healthy female volunteers were randomly divided into four groups, namely control group, experimental group 1, experimental group 2 and experimental group 3, with 30 people in each group. The control group had a normal diet and did not take any additional elastin peptide products. In addition to the normal diet, each person in the experimental group 1 orally added 3g of a commercially available elastin peptide every day, with the contents of desmosin and isodesmosin being 0.07% and 0.13% respectively. Each person in the experimental group 2 orally added 3g of the elastin peptide product prepared in comparative example 3 every day, and each person in the experimental group 3 orally added 3g of the elastin peptide product prepared in example 1 every day, all taken with warm water for a total of 12 weeks. On the 0th day from the beginning of the experiment and the 90th day after the end of the experiment, CORTEX ( Combo) Skin Physiological Index Tester is used to detect the moisture content of the stratum corneum and skin elasticity of the volunteers' skin.

[0086] Table 2 Changes in moisture content of skin stratum corneum before and after the test

[0087] Grouping 0d(μS) 90d(μS) Rate of change (%) Control group 182.50±16.09 193.14±14.15 5.83 Experimental Group 1 189.15±19.13 235.03±20.01 24.26 Experimental Group 2 190.50±21.07 233.33±24.61 22.48 Experimental Group 3 198.50±20.11 276.30±29.90 39.19

[0088] As can be seen from Table 2, the subjects' skin moisture content was improved after taking the commercially available elastin peptide and the elastin peptide with lower purity prepared in Comparative Example 3, and the moisture content of the stratum corneum of the subjects' skin was improved more significantly after taking the high-purity elastin peptide prepared in Example 1. This indicates that the high-purity elastin peptide prepared by the method of the present invention has a more significant effect on improving the moisture content of the skin.

[0089] Table 3 Changes in skin elasticity before and after the test

[0090]

[0091]

[0092] As can be seen from Table 3, the subjects' skin elasticity was improved after supplementing with the elastin peptide prepared in Comparative Example 3, while the skin elasticity was improved to a greater extent after taking the elastin peptide with high desmosin and isodesmosin content prepared in Example 1, indicating that the high-purity elastin peptide prepared by the method of the present invention is more conducive to improving and enhancing the elasticity of the skin than the ordinary commercially available elastin peptide, and also indicating that the method of the present invention has higher physical health and socioeconomic value.

[0093] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing low-fat, low-cost, high-purity and high-quality elastin peptides. Features: The preparation method comprises the following steps: (1) Pretreatment: Cut the cleaned elastin-rich animal tissue into small pieces, add 8-12 times the weight of an alkaline solution, soak at room temperature for 2-6 hours, and wash with purified water until the conductivity of the effluent is within 1000 μs / cm, thereby obtaining a pretreated elastin tissue, wherein the alkaline solution is a mixed solution of alkali and sodium carbonate, and the mass concentration ratio of alkali to sodium carbonate is (0.05-0.1%): (0.5-1%); (2) Heating aging: After cleaning, the pretreated elastin tissue prepared in step (1) is added with 1-3 times the weight of purified water, and then the material is heated by microwave heating. The microwave power is constant at 500-800 W. The microwave treatment is performed in an intermittent manner. After working for 2-4 minutes, it is rested for 2-4 minutes. One microwave action and rest is considered as one cycle. 6-8 cycles are performed. (3) Two-phase separation: The liquid after heat extraction is allowed to stand and the two phases are separated, with the oil phase in the upper layer and the water phase in the lower layer. The oil phase is fully removed by liquid separation; (4) Enzymatic hydrolysis: adding 5-15% of the weight of the pretreated elastin tissue to the aqueous phase, adjusting the temperature to 50-65° C., and performing enzymatic hydrolysis for 3-8 hours to obtain an enzymatic hydrolyzate containing elastin peptides; (5) solid-liquid separation: the enzymatic hydrolyzate containing elastin peptides prepared in step (4) is subjected to solid-liquid separation using a 50-100 nm ceramic membrane to obtain a clear and transparent enzymatic hydrolyzate solution; (6) Decolorization and purification: The enzymatic hydrolyzate prepared in step (5) is sequentially passed through a series of chromatography columns filled with a cation exchange resin, anion exchange resin, and cation exchange resin, with a flow rate controlled at 4-6 BV / h, to remove other impurity protein peptides including collagen peptides, and to prepare an elastin peptide solution, wherein the cation exchange resin, anion exchange resin, and cation exchange resin are respectively weak acid type, strong base type, and weak acid type ion exchange resins, and the column loading volume ratio is 1:1.2:1-1:1.5:1; (7) Sterilization: sterilize the elastin peptide solution prepared in step (6) using high voltage pulsed electric field technology (PEF), with an electric field strength of 15-30 KV / cm and a pulse number of 100-350; (8) Drying: spray drying the sterilized elastin peptide solution prepared in step (7) to obtain elastin peptide powder.

2. The preparation method according to claim 1, Features: In step (1), the elastin-rich animal tissue is bovine or porcine aorta, skin, ligament or fish heart artery bulb.

3. The preparation method according to claim 1, Features: In step (1), the base is sodium hydroxide or potassium hydroxide.

4. The preparation method according to claim 1, Features: In step (4), the immobilized protease is selected from one or more of the following: alkaline protease, papain, bromelain, neutral protease, flavor protease, trypsin or pepsin.

5. The preparation method according to claim 4, Features: The immobilized protease is selected from alkaline protease and papain.

6. The preparation method according to claim 1, Features: The preparation method of the immobilized protease in step (4) comprises the following steps: based on the total mass of the immobilized protease, the mass concentration of sodium alginate is 2-4%, the mass concentration of chitosan is 3-5%, the mass concentration of calcium chloride is 2-4%, the mass concentration of glutaraldehyde is 0.5-1%, the mass concentration of protease is 0.5-0.8%, the cross-linking time is 1-4h, and the immobilization temperature is 35-45°C, so as to prepare the microsphere immobilized protease.

7. The preparation method according to claim 1, Features: The method for recovering the immobilized protease in step (4) comprises the following steps: passing the enzymatic solution containing the immobilized protease through a 40-60 mesh standard sieve to recover the microsphere immobilized protease, and rinsing with pure water and refrigerating for repeated use.

8. Elastin peptide prepared by the preparation method according to any one of claims 1 to 7.

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