Defatted sea cucumber peptide and application thereof in preparation of antioxidant sea cucumber peptide solid beverage

Abstract

The present application relates to the technical field of defatting of marine oligopeptides, and in particular to a defatted sea cucumber peptide and its application in preparation of an antioxidant sea cucumber peptide solid beverage. Small molecular oligopeptides are obtained from sea cucumber through directional enzyme cutting, defatting and membrane filtration technology. The antioxidant sea cucumber peptide solid beverage prepared from the sea cucumber peptide has significant antioxidant effect and obvious dose-dependent effect, and is free of fat, stable in shelf life, and good in flowability and solubility.

Description

Technical Field

[0001] This invention relates to the field of marine biological oligopeptide defatting technology, specifically to a defatted sea cucumber peptide and its application in the preparation of antioxidant sea cucumber peptide solid beverages. Background Technology

[0002] Reactive oxygen species (ROS) in living organisms mainly include hydroxyl radicals, superoxide anions, hydrogen peroxide, and nitric oxide, which have strong oxidizing activity. During normal physiological metabolism, ROS are maintained in dynamic balance under the action of superoxide dismutase (SOD), catalase, glutathione peroxidase, and ascorbic acid, and do not cause damage to the body. However, if this balance is disrupted, ROS accumulate in large quantities in the body. Excessive ROS can cause oxidative stress and interfere with normal physiological functions, such as protein structure modification, DNA damage, abnormal cell signaling, reduced proliferation, and host defense defects.

[0003] To combat oxidative stress in the body, numerous researchers have isolated and extracted substances with antioxidant activity, such as plant polysaccharides, probiotics, and bioactive peptides. Considering the body's basic nutritional needs, bioactive peptides show promising application prospects.

[0004] Recent scientific findings suggest that the human body primarily absorbs protein in the form of peptides. Peptides are molecules composed of amino acids, with molecular weights ranging from 5000 to 200 Da. They possess extremely high activity and diversity, and can regulate the physiological functions of various systems and cells in the human body and activate related enzymes through the physiological functions and biological activities of peptide nutrition. Therefore, various peptides have become important substances for human health.

[0005] Sea cucumbers are marine invertebrates belonging to the phylum Echinodermata and class Holothuroidea. Due to their extremely high nutritional and medicinal value, they were known as "ginseng of the sea" in ancient China and were used for consumption as early as 200 AD. The *Compendium of Materia Medica Supplement* records: "Sea cucumbers are sweet and salty in taste, nourish the kidneys, replenish essence, and control urination. Their warm and nourishing nature is comparable to ginseng, hence the name 'sea ginseng.'" Sea cucumbers contain 40.7%–63.6% protein, with collagen accounting for 70% of the total protein. They are also rich in polysaccharides, saponins, trace elements such as calcium, magnesium, and copper, and small amounts of nucleic acids, making them one of the eight treasures of Chinese cuisine. In recent decades, various substances with important biological activities have been discovered in sea cucumbers, including sea cucumber peptides, sea cucumber polysaccharides, sea cucumber saponins, cerebrosides, sphingosine, and phenolic substances, which possess various physiological functions such as antioxidation, anti-inflammation, anti-cancer, antithrombosis, anti-diabetic, anti-obesity, antibacterial, and liver protection. Because bioactive peptides have advantages such as wide availability, mild properties, easy digestion and absorption, diverse activities and extremely low side effects, their extraction and purification processes and biological functions have become a hot topic in the food industry. Moreover, low molecular weight peptides have higher activity, stability and specificity compared to other bioactive substances.

[0006] Sea cucumber peptides refer to protein hydrolysates obtained by separating and purifying sea cucumbers through protease hydrolysis. These hydrolysates are mainly composed of small-molecule peptides and possess various functional properties. They exhibit good solubility, stability, emulsification, and low viscosity, and their structure is not destroyed by proteases or acidic / alkaline substances in the digestive system. They can be fully absorbed and utilized by the human body, thus possessing great application potential. However, in the industrial production of sea cucumber peptides and their products, the raw sea cucumber itself contains a certain amount of fat. If this fat is not removed during the process, it will adversely affect the flavor and shelf life of the raw sea cucumber peptides and their products, thereby impacting the industrial market for sea cucumber peptides and their products. The traditional degreasing method is centrifugal degreasing, as described in patent CN113832206A, a method for preparing sea cucumber peptides, which uses a three-phase centrifuge for degreasing. However, the separation effect of the centrifuge is affected by various factors, such as the mass, size, and density of the particles, as well as the density difference between solids and liquids or liquids and liquids. If these factors are not met, the centrifugal degreasing effect may be unsatisfactory, resulting in products still containing high levels of fat or other impurities and high energy consumption. This invention is based on the existing workshop production line equipment and, according to the principle that the denaturation and gelatinization of proteins and starches cause changes in the rheological and mesoscopic properties of the green "degreasing agent", it achieves simultaneous enzyme inactivation and degreasing, thereby reducing energy consumption while achieving degreasing. Summary of the Invention

[0007] The technical problem to be solved by this invention is that sea cucumber raw materials contain a certain amount of fat. If it is not removed during the process, it will have an adverse effect on the flavor and shelf life of sea cucumber peptide raw materials and their products, thereby affecting the industrial market of sea cucumber peptides and their products.

[0008] To address the problems of existing technologies, this invention discloses a defatted sea cucumber peptide and its application in the preparation of an antioxidant sea cucumber peptide solid beverage. Using sea cucumber as raw material, small molecule oligopeptides are obtained through directional enzymatic hydrolysis, defatting, and membrane filtration. Flour is used as a green "defatting agent" to absorb the fat. Based on the denaturation and gelatinization of proteins and starches, the rheological and mesoscopic properties of the green "defatting agent" change, achieving simultaneous enzyme inactivation and defatting, thus reducing energy consumption while achieving defatting. The prepared antioxidant sea cucumber peptide solid beverage exhibits significant antioxidant effects with a clear dose-dependent effect; it is also fat-free, has a stable shelf life, and excellent flowability and solubility.

[0009] To achieve the above objectives, the present invention specifically implements the following technical solution: a method for preparing defatted sea cucumber peptides, comprising the following steps:

[0010] (0-1) Pre-treatment: mince and grind the sea cucumber;

[0011] (0-2) Enzymatic hydrolysis: After the temperature is constant, add pre-activated animal protease, papain, and neutral protease, with each enzyme added at 0.1-0.3% of the weight of the sea cucumber pulp. Animal protease specifically hydrolyzes proteins in the sea cucumber body wall; papain is used for the hydrolysis of fresh sea cucumber, exhibiting good hydrolysis effects and aiding in the extraction of active ingredients; neutral protease has high hydrolysis efficiency, helping to extract active ingredients from sea cucumber and also producing enzymatic hydrolysates with various physiological activities. Stir and hydrolyze for 4-6 hours. After 4 hours of hydrolysis, measure the solid content every half hour using a solids analyzer. Hydrolysis is considered complete when the solid content is basically stable.

[0012] (0-3) Fat Absorption and Enzyme Inactivation: Add pre-weighed flour at a rate of 0.2-0.5% of the sea cucumber enzymatic hydrolysate weight. Dissolve the flour in warm water, then pour the dissolved flour solution into the enzymatic hydrolysis tank. Stir for 5 minutes, then turn off the stirrer and fully open the steam valve. Heat the hydrolyzed semi-finished product to 98℃ and maintain this temperature for 20 minutes to denature, absorb fat, and inactivate enzymes. Close the steam valve of the enzymatic hydrolysis tank and add ice water to cool it down to below 40℃. Open the discharge valve, and pass the cooled semi-finished product through a 100-mesh sieve to remove sand and other non-enzymatically hydrolyzed residues. Transfer the hydrolysate through the sieve into a temporary storage tank. Disperse the starch in the flour solution with water, then heat and disperse it into the sea cucumber enzymatic hydrolysate. Heat the entire solution. At 50℃, the flour solution begins to absorb water and swell. When heated to 65℃, protein denaturation is observed, and white flocculent matter appears in the enzymatic hydrolysate. Further heating to 98℃-100℃ causes the hydrolysate to begin boiling, and the white flocculent matter aggregates and floats on top, turning yellow and becoming smooth. During this process, the starch in the flour solution swells and gelatinizes, increasing in volume and dissolving in water. When added to the lipid-containing enzymatic hydrolysate, the fat enters the starch solution micelles, forming a lipid sol. At this point, the heated starch blocks clearly turn yellow. Simultaneously, the protein in the flour solution swells upon contact with water, increasing its surface area and adsorbing fat from the enzymatic hydrolysate during the denaturation process.

[0013] (0-4) Separation: The cooled semi-finished product is separated in a tubular centrifuge at 4000 r / min and a feed rate of 1.0 m / s. 3 / h, the insoluble residue of 100 mesh or more is retained. The enzymatic hydrolysate filtrate is then filtered again through a plate and frame filter at 0.8 MPa and 22 plates to retain the remaining insoluble residue (300 mesh or more). The filtrate is then transferred to a temporary storage tank.

[0014] (0-5) Membrane filtration: The semi-finished product in the temporary storage tank is filtered through a 10,000 Dalton membrane to obtain an enzymatic hydrolysate with a molecular weight lower than 10,000 Daltons. The filtrate is collected into the temporary storage tank;

[0015] (0-6) Concentration: The filtrate in the temporary storage tank is pumped to a double-effect vacuum concentrator for concentration. The evaporation pressure is 0.06 MPa, and the evaporation temperature is 68-70℃ for the first effect and 56-60℃ for the second effect. After the filtrate is concentrated to a solid content of 15%-20%, it is injected into the mixing tank through a pipeline for later use.

[0016] (0-7) Drying: Drying yields defatted sea cucumber peptide powder.

[0017] Further, in step (0-1), the frozen sea cucumber plate is first passed through a frozen plate meat grinder and then connected to a colloid mill. After being ground, it is pumped through a vacuum pump to the feed port of the colloid mill and water is added at a ratio of 2:1 while controlling the flow rate. After passing through the colloid mill, it is pumped into an enzymatic hydrolysis tank. The steam valve in the jacket of the enzymatic hydrolysis tank is opened to raise the temperature and start stirring. The temperature is set to 56±2℃.

[0018] Furthermore, steps (0-7) employ spray drying. The specific steps are as follows: after the concentrated enzymatic hydrolysate is homogenized, it is fed into a spray tower for spray drying. The inlet temperature of the spray tower is 170-190℃, and the outlet temperature is 95-105℃. A vibrating screen is installed at the powder outlet, and the screen is sieved once every 8-12 minutes to collect defatted sea cucumber peptide powder.

[0019] A defatted sea cucumber peptide prepared by the above method.

[0020] The above-mentioned defatted sea cucumber peptides are used in the preparation of antioxidant sea cucumber peptide solid beverages, wherein the defatted sea cucumber peptide powder accounts for 5-30% by weight. The recommended daily intake of sea cucumber peptides is 1.0-2.0 grams, and the amount added can be customized according to the packaging specifications of the solid beverage (5-30 grams / bag).

[0021] An antioxidant sea cucumber peptide solid beverage is composed of the following ingredients by weight percentage: 5-30% defatted sea cucumber peptide powder, 1-3% natural coloring agent and antioxidant—astaxanthin microcapsule powder, 10-20% natural flavoring ingredient—apple powder, 5-15% natural flavoring ingredient and coloring ingredient—cranberry fruit powder, 0.05-0.15% sweetener—stevioside, 30%-51% filler to improve product flowability—maltodextrin, 3-10% prebiotic—inulin, 3-7% water-soluble dietary fiber—resistant dextrin, 0.2-1.5% flavoring—food flavoring, and 0.01-0.02% pyrroloquinoline quinone. It provides a systematic antioxidant effect from multiple perspectives, including a significant reduction in malondialdehyde (MDA) and protein carbonyl content, and a significant increase in superoxide dismutase (SOD) activity and reduced glutathione (GSH) content.

[0022] The preparation method of the above-mentioned antioxidant sea cucumber peptide solid beverage includes the following steps:

[0023] (1) Weighing and mixing: Weigh each raw material accurately according to the formula dosage, put the weighed raw materials into the three-dimensional mixer, seal it, and mix for 20-40 minutes until uniform.

[0024] (2) Fluidized bed granulation: Turn on the fluidized bed granulation equipment, suck the mixture into the hopper sealed container, granulate, dry, and obtain granules with a moisture content of <5%;

[0025] (3) Granulation: The granulated particles are passed through an 80-mesh sieve to obtain relatively uniform particles;

[0026] (4) Packaging: Put the semi-finished granules into the granule packaging machine, code and package them;

[0027] (5) Packaging: Pack the pre-packaged solid beverage into a coded box and seal it with a cigarette after packaging.

[0028] Furthermore, step (1) also includes unpacking: before each raw material enters the weighing room, the outer packaging bag is removed and replaced with a sterile bag specifically for the workshop and marked.

[0029] The beneficial effects of this invention are as follows:

[0030] (1) The obtained sea cucumber peptide has a molecular weight of <10000Da, high purity, no fat or other raw materials, good palatability, solubility and flowability, stable shelf life and wide range of applications.

[0031] (2) Compared with the prior art, the composition of the present invention is reasonably matched, ensuring the content of sea cucumber peptides while having good palatability. It has the characteristics of zero added sucrose and no fat. It has a systematic antioxidant effect based on indicators such as the content of lipid oxidation product MDA, protein carbonyl content, reduced GSH content, antioxidant enzyme SOD and GSH-Px activity, which can play a significant role in preventing the body from peroxidation and has an auxiliary role in improving the quality of life. Attached Figure Description

[0032] Figure 1 This is a flowchart of the preparation method of the antioxidant defatted sea cucumber peptide solid beverage of the present invention. Detailed Implementation

[0033] The following description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

[0034] The following embodiments can be understood as illustrating only a part of the structure or method of the present invention, or as a combination of embodiments explaining the broader structure or method of the present invention. Unless otherwise specified, all materials used in the present invention are commercially available.

[0035] Example 1:

[0036] A method for preparing defatted sea cucumber peptides includes the following steps:

[0037] (0-1) Pretreatment: The frozen sea cucumber plates are first passed through a frozen plate meat grinder and then connected to a colloid mill. After being ground, they are pumped to the feed port of the colloid mill by a vacuum pump and water is added at a ratio of 2:1. At the same time, the flow rate is controlled. After passing through the colloid mill, they are pumped into an enzymatic hydrolysis tank. The steam valve in the jacket of the enzymatic hydrolysis tank is opened to raise the temperature and start stirring. The temperature is set to 56±2℃.

[0038] (0-2) Enzymatic hydrolysis: After the temperature is constant, add animal protease, papain and neutral protease that have been activated in warm water in advance. The amount of each enzyme preparation added is 0.2% of the weight of the sea cucumber hydrolysate. Stir and hydrolyze for 4-6 hours. After 4 hours of hydrolysis, measure the solid content every half hour with a solid content analyzer. When the solid content is basically stable, the hydrolysis is over.

[0039] (0-3) Fat absorption and enzyme inactivation: Add 0.2% of the weight of the sea cucumber enzymatic hydrolysate to the pre-weighed flour and dissolve it in warm water. Put the dissolved flour solution into the enzymatic hydrolysis tank, stir and mix for 5 minutes, turn off the stirring, and fully open the steam valve. Heat the enzymatically hydrolyzed semi-finished product to 98°C and maintain it for 20 minutes to denature, absorb fat, and inactivate enzymes. Close the steam valve of the enzymatic hydrolysis tank and pump in ice water to cool it down to below 40°C. Open the discharge valve and pump the cooled semi-finished product through a 100-mesh sieve into the temporary storage tank.

[0040] (0-4) Separation: The cooled semi-finished product is separated in a tubular centrifuge at 4000 r / min and a feed rate of 1.0 m / s. 3 / h, the insoluble residue of 100 mesh or more is retained. The enzymatic hydrolysate filtrate is then filtered again through a plate and frame filter at 0.8 MPa and 22 plates to retain the remaining insoluble residue (300 mesh or more). The filtrate is then transferred to a temporary storage tank.

[0041] (0-5) Membrane filtration: The semi-finished product in the temporary storage tank is filtered through a 10,000 Dalton membrane using a membrane filtration device, and the filtrate is collected and stored in the temporary storage tank;

[0042] (0-6) Concentration: The filtrate in the temporary storage tank is pumped to a double-effect vacuum concentrator for concentration. The evaporation pressure is 0.06 MPa, and the evaporation temperature is 68-70℃ for the first effect and 56-60℃ for the second effect. After the filtrate is concentrated to a solid content of 15%-20%, it is injected into the mixing tank through a pipeline for later use.

[0043] (0-7) Spray drying: After the concentrated enzymatic hydrolysate is homogenized, it is spray dried in a spray tower. The inlet temperature of the spray drying tower is 170-190℃ and the outlet temperature is 95-105℃. A vibrating screen is set at the powder outlet, and the screen is sieved once every 8-12 minutes to collect defatted sea cucumber peptide powder.

[0044]

[0045] According to GB5009.6-2016 "National Food Safety Standard - Determination of Fat in Food - Soxhlet Extraction Method", the fat content of two batches of sea cucumber peptide powder (without defatting process and with flour defatting process) was tested. In one batch, the fat content (wet basis) of the sea cucumber peptide powder without defatting process was 5.63%, and the fat content (dry basis) was 5.88%. The fat content (wet basis) of the sea cucumber peptide powder with flour defatting process was 1.36%, and the fat content (dry basis) was 1.4%. In the other batch, the fat content (wet basis) of the sea cucumber peptide powder without defatting process was 3.63%, and the fat content (dry basis) was 3.74%. The fat content (wet basis) of the sea cucumber peptide powder with flour defatting process was 0.21%, and the fat content (dry basis) was 0.23%. Based on the results of the defatted sea cucumber peptide powder results, the fat content of the sea cucumber peptide powder with flour defatting process ranges from 0.2% to 1.4%.

[0046] Example 2:

[0047] An antioxidant defatted sea cucumber peptide solid beverage is composed of the following ingredients in weight percentage: 10% defatted sea cucumber peptide powder from Example 1, 1.5% astaxanthin microcapsule powder, 17% apple powder, 10% cranberry fruit powder, 0.1% steviol glycosides, 50.39% maltodextrin, 5% inulin, 5% resistant dextrin, 1% edible flavoring, and 0.01% pyrroloquinoline quinone.

[0048] Reference Figure 1 As shown, a method for preparing an antioxidant defatted sea cucumber peptide solid beverage is described below:

[0049] (1) Unpacking, weighing, and mixing: Before entering the weighing room, remove the outer packaging bags of each raw material and replace them with sterile bags specifically for the workshop and label them. Accurately weigh each raw material according to the formula dosage, put the weighed raw materials into the three-dimensional mixer, seal it, and mix for 20-40 minutes until uniform;

[0050] (2) Fluidized bed granulation: Turn on the fluidized bed granulation equipment, suck the mixture into the hopper sealed container, granulate, dry, and obtain granules with a moisture content of <5%;

[0051] (3) Granulation: The granulated particles are passed through an 80-mesh sieve to obtain relatively uniform particles;

[0052] (4) Packaging: Put the semi-finished granules into the granule packaging machine, code and package them;

[0053] (5) Packaging: Pack the pre-packaged solid beverage into a coded box and seal it with a cigarette after packaging.

[0054] The antioxidant defatted sea cucumber peptide solid beverage prepared above was applied to animals in terms of oxidative stimulation, and the experimental process and results are described as follows.

[0055] 1. Experimental animals

[0056] SPF-grade male Kunming mice provided by Beijing Vital River Laboratory Animal Technology Co., Ltd., weighing about 25 g, were selected. The production license number of experimental animals is SYXK(Beijing)2022-0052. The experimental environmental temperature was 22±2°C, and the relative humidity was 50±10%. The experimental animal use license number is SCXK(Shandong)20200022. According to the body weight of the mice, they were randomly divided into 5 groups: model control group, low-dose group, medium-dose group, high-dose group and blank control group, with 8 mice in each group.

[0057] 2. Experimental grouping

[0058] Table 1 Gavage dose and grouping

[0059]

[0060] 3. Animal treatment

[0061] After the last gavage, except for the blank control group, the other four groups of mice were fasted for 16 h, and then gavaged once with 50% ethanol (calculated at 12 mL / kg·bw). After 6 h, blood samples were taken from the orbital cavity (the blank control group was not treated and the samples were taken without fasting), and the contents of lipid oxidation product MDA, protein carbonyl, reduced GSH, and the activities of antioxidant enzymes SOD and GSH-Px in the blood were measured.

[0062] 4. Experimental results

[0063] 4.1. Effect of antioxidant defatted sea cucumber peptide solid beverage on the body weight of mice

[0064] Table 1 Effect of antioxidant defatted sea cucumber peptide solid beverage on the body weight of mice

[0065]

[0066]

[0067] As can be seen from Table 1, there were no significant differences in the body weights of the mice in the model control group and each dose group of the antioxidant defatted sea cucumber peptide solid beverage compared with the blank control group, and there was no statistical significance (P>0.05).

[0068] 4.2. Effect of antioxidant defatted sea cucumber peptide solid beverage on the contents of peroxidized lipid and protein carbonyl in the blood of mice

[0069] Table 2. Effects of antioxidant defatted sea cucumber peptide solid beverage on malondialdehyde content and protein carbonyl content in mouse blood.

[0070]

[0071] Compared with the model control group, * P<0.05, ** P<0.01

[0072] As shown in Table 2, the levels of lipid peroxidation (MDA) and protein carbonyl groups in the blood of mice in the high-dose group of the antioxidant defatted sea cucumber peptide solid beverage were significantly lower than those in the model control group (P<0.05). This indicates that the antioxidant defatted sea cucumber peptide solid beverage can reduce oxidative stress in mice, suggesting that the high-dose group of mice has enhanced antioxidant capacity and reduced oxidative damage. 4.3 Effects of the antioxidant defatted sea cucumber peptide solid beverage on the activity of antioxidant enzymes and the content of reduced glutathione in the blood of mice.

[0073] Table 3. Effects of antioxidant defatted sea cucumber peptide solid beverage on antioxidant enzyme activity and reduced glutathione content in mouse serum.

[0074]

[0075] Compared with the model control group, * P<0.05, ** P<0.01

[0076] As shown in Table 3, the SOD activity and reduced GSH content of erythrocytes in the high-dose group of the antioxidant defatted sea cucumber peptide solid beverage were significantly higher than those in the model control group, which was statistically significant (P<0.05). This indicates that the antioxidant defatted sea cucumber peptide solid beverage can enhance the antioxidant capacity of mice, reduce oxidative damage, and enhance the body's antioxidant defense mechanism, thereby protecting cells from oxidative damage.

[0077] This experiment showed that oral administration of defatted sea cucumber peptide solid beverage samples to mice at doses of 0.833 g / kg·bw, 1.67 g / kg·bw, and 5.01 g / kg·bw for 30 consecutive days had no adverse effect on the weight gain of the mice. Compared with the model control group, the high-dose group of defatted sea cucumber peptide solid beverage showed significantly lower levels of lipid peroxidation MDA and protein carbonyl groups in the blood, and significantly increased SOD activity and reduced glutathione content. Based on the animal experimental criteria for determining antioxidant function in the "Evaluation Methods for Functions of Health Foods (2020 Edition) Draft for Comments," this defatted sea cucumber peptide solid beverage can be considered to have antioxidant function.

[0078] All aspects, embodiments, and features of this invention should be considered illustrative in all respects and not limiting of the invention; the scope of the invention is defined only by the claims. Other embodiments, modifications, and uses will become apparent to those skilled in the art without departing from the spirit and scope of the invention as claimed.

[0079] In the preparation method of this invention, the order of the steps is not limited to the listed order. For those skilled in the art, variations in the order of the steps without creative effort are also within the scope of protection of this invention. Furthermore, two or more steps or actions can be performed simultaneously.

[0080] Finally, it should be noted that the specific embodiments described herein are merely illustrative examples of the invention and are not intended to limit the implementation of the invention. Those skilled in the art can make various modifications or additions to the described specific embodiments or use similar methods to replace them; it is neither necessary nor possible to exemplify all embodiments here. However, these obvious variations or modifications derived from the essential spirit of the invention still fall within the scope of protection of the invention, and interpreting them as any additional limitation would contradict the spirit of the invention.

Claims

1. A method for preparing defatted sea cucumber peptide, characterized in that Comprising the following steps: (0-1) Pretreatment: sea cucumber is minced and polished; (0-2) Enzymatic hydrolysis: after constant temperature, animal protease, papain and neutral protease activated in advance by warm water are added, and the enzyme preparation is added in an amount of 0-0.3% of the weight of the polished sea cucumber slurry, and the enzymatic hydrolysis is stirred for 4-6 hours. After 4 hours of enzymatic hydrolysis, the solid content is measured every half hour with a solid content meter. When the solid content is stable, the enzymatic hydrolysis is completed; (0-3) Fat absorption and enzyme inactivation: the weighed flour is added to the enzymatic hydrolysis tank, and the amount of flour added is 0.2-0.5% of the weight of the sea cucumber enzymatic hydrolysis solution. The flour solution is dissolved with warm water, and then stirred for 5 minutes. The stirring is turned off, the steam valve is fully opened, and the enzymatic hydrolysis product is heated to 98℃ for 20 minutes to denature, absorb fat and inactivate the enzyme. The steam valve of the enzymatic hydrolysis tank is closed, and ice water is injected to cool it to below 40℃. The lower discharge valve is opened, and the cooled product is discharged through a 100-mesh screen into a temporary storage tank; (0-4) separation: the semi-product link tube centrifuge is separated after cooling, the filtrate is passed through the plate and frame filter equipment again and then is connected into the temporary storage tank; the centrifugal parameter is 4000 r / min, the feeding speed is 1.0 m / h, the insoluble residue above 100 meshes is intercepted, the obtained enzymatic hydrolysate filtrate is passed through the plate and frame filter equipment again, 0.8 Mpa, 22 plates, the remaining insoluble residue above 300 meshes is intercepted, and the filtrate is connected into the temporary storage tank; 3 ​ (0-5) Membrane filtration: the product in the temporary storage tank is filtered through a 10,000-dalton membrane, and the filtrate is collected in a temporary storage tank; (0-6) Concentration: The filtrate in the temporary storage tank is concentrated in a double-effect vacuum concentration device. When the filtrate is concentrated to 15-20% solid content, it is injected into a blending tank through a pipeline for later use; (0-7) Drying: The dried defatted sea cucumber peptide powder is obtained.

2. The production method according to claim 1, characterized by: Step (0-1): The sea cucumber frozen plate is first passed through the frozen plate meat grinder and then connected to the colloid mill. After mincing, it is pumped into the colloid mill inlet with 2:1 water addition and controlled flow rate. After passing through the colloid mill, it is pumped into the enzymatic hydrolysis tank. The steam valve in the enzymatic hydrolysis tank is opened for heating, and the stirring is started. The temperature is set to 56±2℃.

3. The production method according to claim 1, wherein: The concentration parameters in step (0-6) are evaporation pressure 0.06Mpa, evaporation temperature, one-effect 68-70℃, and two-effect 56-60℃.

4. The production method according to claim 1, wherein: Step (0-7) uses spray drying. The specific steps are as follows: the concentrated enzymatic hydrolysate is passed through a homogenizer and then enters the spray tower for spray drying. The inlet temperature of the spray tower for spray drying is 170-190℃, and the outlet temperature is 95-105℃. The powder outlet is equipped with a vibrating screen, which is sieved once every 8-12 minutes, and the defatted sea cucumber peptide powder is collected.

5. The defatted sea cucumber peptide prepared by any of the methods of claims 1-4.

6. The use of the defatted sea cucumber peptide of claim 5 in the preparation of an antioxidant sea cucumber peptide solid beverage, characterized in that: The weight of the defatted sea cucumber peptide powder in the solid beverage is 5-30%.

7. An antioxidant sea cucumber peptide solid beverage, characterized in that The defatted sea cucumber peptide powder of claim 5, 5-30% astaxanthin microcapsule powder, 10-20% apple powder, 5-15% cranberry fruit powder, 0.05-0.15% steviol glycoside, 30%-51% malt dextrin, 3-10% inulin, 3-7% resistant dextrin, 0.2-1.5% food flavor, and 0.01-0.02% pyrroloquinoline quinone.

8. Process for the preparation of a solid beverage according to claim 7, characterized in that Comprising the following steps: (1) Weighing and mixing: according to the formula, accurately weigh each raw material, and put the weighed raw materials into a three-dimensional mixer, close it, and mix for 20-40 minutes until uniform; (2) Boiling granulation: open the boiling granulation equipment, and suck the mixed material into the closed container in the hopper to form particles, dry, and obtain granules with a moisture content of <5%; (3) whole grain: the granulation of good particles through 80 mesh screen, to obtain more uniform particles; (4) subpackaging: the granular semi-product is put into a granular subpackaging machine, and is coded and subpackaged; (5) packaging: the subpackaged solid beverage is put into a coded packaging box, and is sealed after the boxing is completed.

9. The production method according to claim 8, characterized by: Step (1) further comprises unpacking: each raw material is unpacked before entering the weighing room, and a workshop special sterile bag is used and marked.

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