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

Abstract

The invention relates to the technical field of marine organism oligopeptide degreasing, in particular to a degreased sea cucumber peptide and application thereof in preparation of an antioxidant sea cucumber peptide solid beverage. The small-molecular oligopeptide is obtained by taking sea cucumber as a raw material and carrying out directional enzyme digestion, degreasing and membrane filtration technologies. The antioxidative sea cucumber peptide solid beverage prepared from the sea cucumber peptide can obviously resist oxidation and has an obvious dose-dependent effect; the product does not contain fat, and is stable in shelf life, and good in fluidity and dissolvability.

Description

Technical Field

[0001] The invention relates to the technical field of marine oligopeptide defatting, and in particular to a defatted sea cucumber peptide and application thereof in the preparation of an antioxidant sea cucumber peptide solid beverage. Background Art

[0002] Reactive oxygen species (ROS) in organisms mainly include hydroxyl free radicals, superoxide anions, hydrogen peroxide and nitric oxide, which have strong oxidative activity. In the normal physiological metabolism of the human body, ROS will maintain a dynamic balance under the action of superoxide dismutase (SOD), catalase, glutathione enzyme and ascorbic acid, and will not cause damage to the human body. However, if this balance is broken, ROS will accumulate in the body in large quantities. Excessive ROS will cause oxidative stress and interfere with the body's normal physiological functions, such as protein structure modification, DNA damage, abnormal cell signaling, reduced proliferation response and host defense defects.

[0003] In order to deal with the body's peroxidation, a large number of researchers have isolated and extracted substances such as plant polysaccharides, probiotics and active peptides with antioxidant activity. Considering the basic nutritional supplementation of the human body, active peptides have a good application prospect.

[0004] In recent years, science believes that the human body absorbs protein mainly in the form of peptides. Peptides are molecules composed of amino acids with a molecular weight between 5000 and 200 Da. They have extremely strong activity and diversity. They can use the physiological functions and biological activities of peptide nutrition to regulate the physiological functions of various systems and cells in the human body and activate related enzymes in the body. Therefore, various peptides have become important substances for human health.

[0005] Sea cucumber is a marine invertebrate belonging to the phylum Echinodermata and class Holothuroidea. Because of its extremely high nutritional and medicinal value, it was called "sea ginseng" in ancient China and was used for food as early as 200 years ago. It is recorded in "Supplement to Compendium of Materia Medica": Sea cucumber tastes sweet and salty, nourishes the kidney, benefits the essence, and controls urination. It is warm and nourishing, and is as good as ginseng, so it is called sea cucumber. The protein content in sea cucumber is as high as 40.7% to 63.6%, of which collagen accounts for 70% of the total protein. It is also rich in polysaccharides and saponins, trace elements such as calcium, magnesium, copper, and a small amount of nucleic acid, and is listed as the first of the eight delicacies. In recent decades, a variety of substances with important biological activities have been found in sea cucumbers, including sea cucumber peptides, sea cucumber polysaccharides, sea cucumber saponins, cerebrosides, sphingosine, phenolic substances, etc., which have multiple physiological functions such as antioxidant, anti-inflammatory, anti-cancer, anti-thrombotic, anti-diabetic, anti-obesity, antibacterial and liver protection. Because bioactive peptides have the advantages of wide sources, 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 food research. Low molecular weight peptides have higher activity, stability and specificity than other bioactive substances.

[0006] Sea cucumber peptide refers to a protein hydrolyzate obtained by separating and purifying sea cucumber after protease hydrolysis, which is mainly composed of small molecule peptides and has multiple functional properties. It has good solubility, stability, emulsification and low viscosity, and its structure will not be destroyed by proteases and acidic and alkaline substances in the digestive system. It can be fully absorbed and utilized by the human body, so it has great application potential. However, in the industrial production of sea cucumber peptides and their products, because the sea cucumber raw materials themselves contain a certain amount of fat, if it is not removed in the process, it will have an adverse effect on the flavor and shelf life of the sea cucumber peptide raw materials and their products, thereby affecting the industrial market of sea cucumber peptides and their products. The traditional degreasing method is centrifugal degreasing, as described in the preparation method of a sea cucumber peptide in patent CN113832206A, a three-phase centrifuge is used for degreasing, but the separation effect of the centrifuge is affected by many factors, such as the quality, size and density of the particles, and the density difference between solid and liquid or liquid and liquid. If these factors do not meet the requirements, the effect of centrifugal degreasing may be unsatisfactory, resulting in the product still containing high fat or other impurities and high energy consumption. The present invention is based on the existing workshop production line equipment conditions, and according to the principle that the rheological properties and mesoscopic properties of the green "degreasing agent" change due to the denaturation and gelatinization of protein and starch, thereby achieving simultaneous enzyme inactivation and degreasing, thereby achieving degreasing and reducing energy consumption. Summary of the invention

[0007] The technical problem to be solved by the present invention is that the sea cucumber raw material itself contains 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 the sea cucumber peptide raw material and its products, thereby affecting the industrial market of sea cucumber peptides and their products.

[0008] In view of the problems of the prior art, the present invention discloses a defatted sea cucumber peptide and its application in the preparation of an antioxidant sea cucumber peptide solid beverage. Small molecule oligopeptides are obtained using sea cucumber as raw material through directional enzyme cutting, defatting and membrane filtration technology; wherein, flour is used to absorb fat, and flour is used as a green "degreasing agent". According to the denaturation and gelatinization of protein and starch, the rheological properties and mesoscopic properties of the green "degreasing agent" are changed, and enzyme inactivation and defatting are carried out simultaneously, achieving degreasing while reducing energy consumption. The prepared antioxidant sea cucumber peptide solid beverage can significantly resist oxidation and has a significant dose-dependent effect; it is fat-free, has a stable shelf life, and has good fluidity and solubility.

[0009] To achieve the above object, the present invention is specifically implemented by the following technical scheme: A method for preparing defatted sea cucumber peptide comprises the following steps:

[0010] (0-1) Pre-treatment: mincing and grinding of sea cucumbers;

[0011] (0-2) Enzymatic hydrolysis: After the temperature is constant, add animal protease, papain and neutral protease activated by warm water in advance, and the amount of each enzyme preparation added is 0.1-0.3% of the weight of the sea cucumber polished slurry. Among them, animal protease specifically enzymatically hydrolyzes the protein in the sea cucumber body wall; papain is used for the hydrolysis of fresh sea cucumbers, has a good hydrolysis effect, and helps to extract the effective ingredients in sea cucumbers; neutral protease has efficient hydrolysis ability, helps to extract the active ingredients in sea cucumbers, and can also produce enzymatic products with multiple physiological activities. Stir the enzymatic hydrolysis for 4-6 hours. After 4 hours of enzymatic hydrolysis, use a solids analyzer to measure the solid content every half an hour. When the solid content is basically stable, the enzymatic hydrolysis is completed;

[0012] (0-3) Fat absorption and enzyme inactivation: Add 0.2-0.5% 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 hydrolysate tank and stir for 5 minutes. Turn off the stirring and fully open the steam valve. Heat the half-product after enzymatic hydrolysis to 98°C and keep it for 20 minutes to denature, absorb fat and inactivate enzymes. Close the steam valve of the enzymatic hydrolysate tank and pour in ice water to cool it down to below 40°C. Open the lower discharge valve, pass the cooled half-product through a 100-mesh sieve to remove sand and other residues that cannot be hydrolyzed by the enzymatic hydrolysate, and pour the enzymatic hydrolysate through the sieve into a temporary storage tank. The starch in the flour aqueous solution is dispersed with water and heated to disperse into the sea cucumber enzymatic hydrolysate. Heat the whole thing. When the temperature reaches 50℃, the flour solution begins to absorb water and expand. When the whole is heated to 65℃, the protein in the flour solution can be observed to be denatured, and white floccules appear in the hydrolyzate. When the temperature continues to be heated to 98℃-100℃, the hydrolyzate begins to boil, and the white floccules gather and float on the top of the hydrolyzate, turning yellow and smooth. During this process, the starch in the flour solution swells and gelatinizes, increases in volume, and dissolves in water. When added to the fat-containing hydrolyzate, the fat enters the starch solution micelles to form a fat-containing sol. At this time, it can be clearly observed that the color of the starch block after heating turns yellow. At the same time, the protein in the flour solution swells with water to form an area, and absorbs the fat in the hydrolyzate during the temperature rise and denaturation process.

[0013] (0-4) Separation: After cooling, the half-product is separated by a linked tubular centrifuge, 4000r / min, and the feed speed is 1.0m 3 / h, intercepting insoluble residues above 100 mesh, the enzymatic hydrolyzate filtrate is filtered through a plate and frame filter again, 0.8Mpa, 22 plates, intercepting the remaining insoluble residues (above 300 mesh), and the filtrate is transferred to a temporary storage tank;

[0014] (0-5) Membrane filtration: The half-product in the temporary storage tank is filtered through a membrane filtration device and passed through a 10,000 Dalton membrane to obtain an enzymatic hydrolyzate with a molecular weight less than 10,000 Daltons. The filtrate is collected in a temporary storage tank;

[0015] (0-6) Concentration: The filtrate in the temporary storage tank is sent to a double-effect vacuum concentration device for concentration, with an evaporation pressure of 0.06 MPa and an evaporation temperature of 68-70°C for the first effect and 56-60°C for the second effect. After the filtrate is concentrated to a solid content of 15%-20%, it is injected into a blending tank through a pipeline for standby use;

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

[0017] Furthermore, 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 minced, it is pumped to the feed port of the colloid mill through a vacuum pump and water is added in a 2:1 ratio while controlling the flow rate. After passing through the colloid mill, it is pumped into an enzymolysis tank. The steam valve in the interlayer of the enzymolysis tank is opened to increase the temperature, stirring is started, and the temperature is set to 56±2°C.

[0018] Furthermore, step (0-7) adopts spray drying, and the specific steps are: the concentrated enzymatic hydrolysate passes through a homogenizer and then enters a spray tower for spray drying, the inlet temperature of the spray tower for spray drying is 170-190°C, and the outlet temperature is 95-105°C; a vibrating screen is set at the powder outlet, and the powder is screened once every 8-12 minutes to collect the defatted sea cucumber peptide powder.

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

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

[0021] An antioxidant sea cucumber peptide solid beverage is composed of the following raw materials in percentage by weight: 5-30% of the above-mentioned defatted sea cucumber peptide powder, 1-3% of natural colorant and antioxidant-astaxanthin microcapsule powder, 10-20% of natural seasoning ingredients-apple powder, 5-15% of natural seasoning ingredients and coloring ingredients-cranberry fruit powder, 0.05-0.15% of sweetener-stevioside, 30%-51% of filler that helps product fluidity-maltodextrin, 3-10% of prebiotic-inulin, 3-7% of water-soluble dietary fiber-resistant dextrin, 0.2-1.5% of flavoring-edible flavor, and 0.01-0.02% of pyrroloquinoline quinone. It has a systematic antioxidant effect from multiple angles, such as significantly reducing the content of lipid peroxidation malondialdehyde (MDA) and protein carbonyl, and significantly increasing the activity of superoxide dismutase (SOD) and the content of reduced glutathione (GSH).

[0022] The method for preparing the above-mentioned antioxidant sea cucumber peptide solid beverage comprises the following steps:

[0023] (1) Weighing and mixing: Accurately weigh the raw materials according to the formula dosage, put the weighed raw materials into a three-dimensional mixer, seal it, and mix it for 20-40 minutes until it is uniform;

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

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

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

[0027] (5) Packaging: The packaged solid beverages are placed in coded packaging boxes and then sealed with cigarettes.

[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 workshop-specific sterile bag and marked.

[0029] The beneficial effects of the present invention are:

[0030] (1) The obtained sea cucumber peptide has a molecular weight of <10,000 Da, high purity, does not contain fat and other raw materials, has good palatability, solubility and fluidity, has a stable shelf life, and has a wide range of applications.

[0031] (2) Compared with the prior art, the composition of the present invention is reasonably matched, and while ensuring the content of sea cucumber peptides, it has good palatability, has the characteristics of zero added sucrose and does not contain fat. It has a systematic antioxidant effect from indicators such as the content of lipid oxidation product MDA, protein carbonyl content, reduced GSH content, antioxidant enzyme SOD and GSH-Px activity, etc., which can play a significant role in preventing the body from peroxidation and has an auxiliary effect on improving the quality of life. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention is a flow chart of a method for preparing an antioxidant defatted sea cucumber peptide solid beverage. DETAILED DESCRIPTION

[0033] The following description is only 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 in the scope of protection of the present invention.

[0034] The following examples can be understood as a part of the local structure or method of the present invention alone, and can also be understood as the combination of the examples to explain the connotation of the larger structure or method of the present invention. Unless otherwise specified, all raw materials of the present invention are obtained from commercial sales.

[0035] Embodiment 1:

[0036] A method for preparing defatted sea cucumber peptide comprises the following steps:

[0037] (0-1) Pretreatment: The frozen sea cucumber plate is first passed through a frozen plate meat grinder and then connected to a colloid mill. After being minced, it is pumped to the feed port of the colloid mill through a vacuum pump and water is added in a ratio of 2:1. The flow rate is controlled at the same time. After passing through the colloid mill, it is pumped into the enzymolysis tank. The steam valve in the interlayer of the enzymolysis tank is opened to increase the temperature, and stirring is started. The temperature is set to 56±2℃;

[0038] (0-2) Enzymolysis: After the temperature is constant, add animal protease, papain and neutral protease activated by warm water in advance, and the amount of each enzyme preparation added is 0.2% of the weight of the sea cucumber enzymolysis solution. Stir and enzymolysis for 4-6 hours. After 4 hours of enzymolysis, the solid content is measured every half an hour with a solids analyzer. When the solid content is basically stable, the enzymolysis ends;

[0039] (0-3) Fat absorption and enzyme inactivation: The flour weighed in advance is added in an amount of 0.2% of the weight of the sea cucumber enzymatic hydrolysate, dissolved in warm water, and the dissolved flour solution is put into the enzymatic hydrolysis tank, stirred and mixed for 5 minutes, the stirring is turned off, the steam valve is fully opened, and the half-product after enzymatic hydrolysis is heated to 98° C. and maintained for 20 minutes for denaturation, fat absorption and enzyme inactivation; the steam valve of the enzymatic hydrolysis tank is closed, and ice water is poured into it for cooling to below 40° C.; the lower discharge valve is opened, and the cooled half-product is poured into the temporary storage tank through a 100-mesh sieve;

[0040] (0-4) Separation: After cooling, the semi-product is separated by a tube centrifuge, 4000r / min, and the feed speed is 1.0m 3 / h, intercepting insoluble residues above 100 mesh, the enzymatic hydrolyzate filtrate is filtered through a plate and frame filter again, 0.8Mpa, 22 plates, intercepting the remaining insoluble residues (above 300 mesh), and the filtrate is transferred to a temporary storage tank;

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

[0042] (0-6) Concentration: The filtrate in the temporary storage tank is sent to a double-effect vacuum concentration device for concentration, with an evaporation pressure of 0.06 MPa and an evaporation temperature of 68-70°C for the first effect and 56-60°C for the second effect. After the filtrate is concentrated to a solid content of 15%-20%, it is injected into a blending tank through a pipeline for standby use;

[0043] (0-7) Spray drying: The concentrated enzymatic hydrolysate is passed through a homogenizer and then into a spray tower for spray drying. The inlet temperature of the spray tower for spray drying is 170-190°C, and the outlet temperature is 95-105°C. A vibrating screen is set at the powder outlet, and the screen is performed once every 8-12 minutes to collect the 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 sea cucumber peptide powder with flour defatting process was tested respectively. The fat content (wet basis) of one batch of 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 sea cucumber peptide powder after flour defatting process was 1.36%, and the fat content (dry basis) was 1.4%; the fat content (wet basis) of the other batch of 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 sea cucumber peptide powder after flour defatting process was 0.21%, and the fat content (dry basis) was 0.23%. Judging from the fat content results of sea cucumber peptide powder after defatting, the fat content of sea cucumber peptide powder after flour defatting process is 0.2-1.4%.

[0046] Embodiment 2:

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

[0048] Reference Figure 1 As shown, a method for preparing an antioxidant defatted sea cucumber peptide solid beverage, the method for preparing the above antioxidant sea cucumber peptide solid beverage is as follows:

[0049] (1) Unpacking, weighing and mixing: Before entering the weighing room, remove the outer packaging bags of each raw material, replace them with sterile bags specially made for the workshop and mark 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 it for 20-40 minutes until it is uniform;

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

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

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

[0053] (5) Packaging: The packaged solid beverages are placed in coded packaging boxes and then sealed with cigarettes.

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

[0055] 1. Experimental Animals

[0056] SPF-grade Kunming male mice provided by Beijing Weitonglihua Experimental Animal Technology Co., Ltd. were selected, weighing about 25g. The experimental animal production license number is SYXK (Beijing) 2022-0052. The experimental environment temperature is 22±2℃, and the relative humidity is 50±10%. The experimental animal use license number is SCXK (Lu) 20200022. Mice were randomly divided into 5 groups according to their weight, including 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 Oral gavage dose and grouping

[0059]

[0060] 3. Animal Handling

[0061] After the last oral administration, except for the blank control group, the other four groups of mice were fasted for 16 h, and then given 50% ethanol (12 mL / / kg·bw) by oral administration once. Blood samples were taken from the eye sockets 6 h later (the blank control group was not treated and samples were collected without fasting) to measure the content of lipid oxidation product MDA, protein carbonyl content, reduced GSH content, and the activities of antioxidant enzymes SOD and GSH-Px in the blood.

[0062] 4. Experimental results

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

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

[0065]

[0066]

[0067] As shown in Table 1, there was no significant difference in the body weight of 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 Effects of antioxidant defatted sea cucumber peptide solid beverage on lipid peroxide content and protein carbonyl content in mouse blood

[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 content of lipid peroxidation MDA and protein carbonyl in the blood of mice in the high-dose group of antioxidant defatted sea cucumber peptide solid beverage was significantly lower than that in the model control group, which was statistically significant (P<0.05). This indicates that the antioxidant defatted sea cucumber peptide solid beverage can reduce oxidative stress in mice, indicating that the antioxidant capacity of mice in the high-dose group is enhanced and oxidative damage is reduced. 4.3. Effect of antioxidant defatted sea cucumber peptide solid beverage on antioxidant enzyme activity and reduced glutathione content in mouse blood

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

[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 antioxidant defatted sea cucumber peptide solid beverage were significantly higher than those in the model control group, with statistical significance (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 antioxidant defense mechanism in the body, thereby protecting cells from oxidative damage.

[0077] This experiment shows that oral administration of 0.833g / kg·bw, 1.67g / kg·bw and 5.01g / kg·bw of defatted sea cucumber peptide solid beverage samples to mice for 30 consecutive days had no adverse effects on the weight gain of mice; compared with the model control group, the high-dose group of defatted sea cucumber peptide solid beverage significantly reduced the levels of lipid peroxidation MDA and protein carbonyl in the blood of mice, and significantly increased the activity of SOD and the content of reduced glutathione. According to the animal experiment judgment criteria for antioxidant function in the "Draft for Comments on the Function Evaluation Method of Health Food (2020 Edition)", this defatted sea cucumber peptide solid beverage can be considered to have antioxidant function.

[0078] The various aspects, embodiments, and features of the present invention should be considered to be illustrative in all aspects and not limiting of the present invention, the scope of which is defined solely by the claims. Other embodiments, modifications, and uses will be apparent to those skilled in the art without departing from the spirit and scope of the claimed invention.

[0079] In the preparation method of the present invention, the order of each step is not limited to the order listed. For those skilled in the art, without creative work, the order of each step is also within the protection scope of the present invention. In addition, two or more steps or actions can be performed simultaneously.

[0080] Finally, it should be noted that the specific embodiments described herein are merely examples of the present invention, and are not intended to limit the implementation methods of the present invention. A person skilled in the art of the present invention may make various modifications or supplements to the specific embodiments described, or replace them in a similar manner. It is not necessary and impossible to provide all examples of all implementation methods here. However, these obvious changes or modifications derived from the essential spirit of the present invention still fall within the scope of protection of the present invention, and interpreting them as any additional limitation is contrary to the spirit of the present invention.

Claims

1. A method for preparing defatted sea cucumber peptide, characterized in that The following steps are involved: (0-1) Pre-treatment: mincing and grinding of sea cucumbers; (0-2) Enzymatic hydrolysis: After the temperature is constant, add animal protease, papain and neutral protease activated by warm water in advance, and the amount of each enzyme preparation added is 0.1-0.3% of the weight of the sea cucumber grinding pulp. Stir and enzymolysis for 4-6 hours. After 4 hours of enzymolysis, the solid content is measured every half an hour with a solids analyzer. When the solid content is basically stable, the enzymolysis ends; (0-3) Fat absorption and enzyme inactivation: The flour weighed in advance is added in an amount of 0.2-0.5% of the weight of the sea cucumber enzymatic hydrolysate, dissolved in warm water, and the dissolved flour solution is put into the enzymatic hydrolysis tank, stirred and mixed for 5 minutes, the stirring is turned off, the steam valve is fully opened, the half-product after enzymatic hydrolysis is heated to 98° C. and maintained for 20 minutes, and denatured fat absorption and enzyme inactivation are performed; the steam valve of the enzymatic hydrolysis tank is closed, and ice water is poured into it to cool it down to below 40° C.; Open the lower discharge valve, and the cooled semi-finished product is put into the temporary storage tank through a 100-mesh screen; (0-4) Separation: After cooling, the semi-product is separated by a tubular centrifuge, and the filtrate is filtered through a plate and frame filter again before being transferred to a temporary storage tank; (0-5) Membrane: The half-product in the temporary storage tank is filtered through a membrane filtration device, passes 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 sent to a double-effect vacuum concentration device for concentration. After the filtrate is concentrated to a solid content of 15%-20%, it is injected into a blending tank through a pipeline for standby use; (0-7) Drying: Drying to obtain defatted sea cucumber peptide powder.

2. The preparation method according to claim 1, characterized in that: 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 minced, it is pumped to the feed port of the colloid mill through a vacuum pump and water is added in a 2:1 ratio while controlling the flow rate. After passing through the colloid mill, it is pumped into an enzymolysis tank. The steam valve in the interlayer of the enzymolysis tank is opened to increase the temperature, stirring is started, and the temperature is set to 56±2°C.

3. The preparation method according to claim 1, characterized in that: The centrifugal parameters in step (0-4) are 4000r / min and the feed speed is 1.0m 3 / h, intercepting insoluble residues above 100 mesh, the enzymatic hydrolyzate filtrate is filtered through the plate and frame filter again, 0.8Mpa, 22 plates, intercepting the remaining insoluble residues above 300 mesh, and the filtrate is connected to the temporary storage tank.

4. The preparation method according to claim 1, characterized in that: The concentration parameters in step (0-6) are evaporation pressure 0.06Mpa, evaporation temperature, 68-70°C for the first effect, and 56-60°C for the second effect.

5. The preparation method according to claim 1, characterized in that: Step (0-7) adopts spray drying, and the specific steps are: the concentrated enzymatic hydrolysate passes through a homogenizer and then enters a spray tower for spray drying, the inlet temperature of the spray tower for spray drying is 170-190°C, and the outlet temperature is 95-105°C; a vibrating screen is set at the powder outlet, and the powder is screened once every 8-12 minutes to collect the defatted sea cucumber peptide powder.

6. The defatted sea cucumber peptide prepared by any method of claims 1-5.

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

8. An antioxidant sea cucumber peptide solid beverage, characterized in that The invention is composed of the following raw materials in percentage by weight: 5-30% of the defatted sea cucumber peptide powder according to claim 6, 1-3% of astaxanthin microcapsule powder, 10-20% of apple powder, 5-15% of cranberry fruit powder, 0.05-0.15% of stevioside, 30%-51% of maltodextrin, 3-10% of inulin, 3-7% of resistant dextrin, 0.2-1.5% of edible flavor and 0.01-0.02% of pyrroloquinoline quinone.

9. The method for preparing the solid beverage according to claim 8, characterized in that The following steps are involved: (1) Weighing and mixing: Accurately weigh the raw materials according to the formula dosage, put the weighed raw materials into a three-dimensional mixer, seal it, and mix it for 20-40 minutes until it is uniform; (2) Fluidized bed granulation: Turn on the fluidized bed granulation equipment, suck the mixed material into a closed container of the hopper, granulate, dry, and obtain granules with a moisture content of <5%; (3) Granulation: The granulated particles are sieved through an 80-mesh sieve to obtain relatively uniform particles; (4) Packaging: Put the semi-finished granules into the granule packing machine, code them, and pack them; (5) Packaging: The packaged solid beverages are placed in coded packaging boxes and then sealed with cigarettes.

10. The preparation method according to claim 9, characterized in that: Step (1) also includes unpacking: before each raw material enters the weighing room, the outer packaging bag is removed and replaced with a workshop-specific sterile bag and marked.

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