Preparation method and application of oyster peptide

Abstract

The invention discloses a preparation method and application of oyster peptide, and belongs to the technical field of marine bioactive substances. The preparation method of the oyster peptide in the oyster peptide energy beverage specifically comprises the following steps: freezing edible parts of fresh oysters, homogenizing, adding 0.05-0.3 wt% of animal compound protease and 0.01-0.2 wt% of neutral protease, and carrying out enzymolysis for 1-7 hours to obtain an oyster peptide crude extract; the oyster peptide crude extract is subjected to decoloration treatment and then filtered, filtrate obtained after filtration is subjected to desalination treatment, the desalted solution is subjected to spray drying treatment, and oyster peptide powder is obtained. According to the invention, the preparation process of the oyster peptide is optimized, the peptide powder with better flavor and better stability in beverage application is obtained, and the problem of fishy smell resistance and precipitation in the application of the oyster peptide in beverage products in the current market is solved.

Description

Technical Field

[0001] The invention belongs to the technical field of marine biological active substances, and in particular, relates to a preparation method of oyster peptide and application thereof. Background Art

[0002] Usually, most people will selectively ignore the sub-healthy state without organic lesions, which will eventually lead to the development of diseases. Therefore, we should pay attention to regulating body functions, getting rid of sub-healthy state, and controlling the occurrence and development of diseases from the source.

[0003] At present, most energy drinks on the market contain taurine, caffeine, guarana, white sugar, vitamins, minerals, etc. Such energy drinks have a good refreshing effect on mature working people and people who are easily fatigued due to high-intensity work, but for students whose physical development is not yet complete, they have a weak ability to metabolize caffeine. A small amount of caffeine is likely to reach a toxic dose and cause harm to them. In addition, the taurine in the formula is basically a synthetic raw material and does not have the biological activity of natural taurine. Studies have shown that taurine is mainly metabolized by the kidneys after being absorbed by the human body to maintain the balance of taurine content in the body. Excessive intake will inevitably cause kidney burden. Regular and large-scale drinking of such beverages is prone to dependence, and the side effects are also obvious. At the same time, a large amount of added white sugar is not suitable for people who lose weight, people with blood sugar regulation dysfunction and people with type 2 diabetes. Therefore, the market urgently needs to launch a new energy drink that can fundamentally solve the above pain points for consumers, cater to market trends, meet consumer needs, have natural and safe raw material components, and have anti-fatigue, anti-oxidation, energy supplement, refreshing and calming functions.

[0004] However, the energy peptide beverage products currently on the market usually have very serious off-flavor and turbidity. The product itself has poor flavor, especially after a shelf life of 8 months. There are usually a large amount of flocculation precipitation visible to the naked eye, and unbearable characteristic flavor and fishy smell. Summary of the invention

[0005] The purpose of the present invention is to provide a preparation method of oyster peptide and application thereof, so as to make up for the deficiencies of the prior art.

[0006] To achieve the above object, the present invention is implemented through the following technical solutions:

[0007] The invention discloses a method for preparing oyster peptides. The method comprises the following steps: enzymolyzing the edible part of oysters, and the enzymes selected are animal composite protease and neutral protease.

[0008] Furthermore, the preparation method is specifically as follows: the edible part of fresh oysters is frozen and then homogenized, 0.05% to 0.3wt% of animal composite protease and 0.01% to 0.2wt% of neutral protease are added, and enzymolysis is performed for 1 to 7 hours to obtain a crude oyster peptide extract; the crude peptide extract is decolorized and then filtered, the filtrate after filtration is desalted, and the desalted solution is spray-dried to obtain oyster peptides.

[0009] Furthermore, the animal composite protease comprises trypsin, pepsin and alkaline protease; the mass fractions of the three enzymes added are: 0.025% to 0.15wt% trypsin, 0.015% to 0.1wt% pepsin, and 0.01% to 0.05wt% alkaline protease.

[0010] Furthermore, the desalination adopts membrane separation technology.

[0011] Application of the oyster peptide in the preparation of beverage products.

[0012] The invention discloses an application of the oyster peptide in the preparation of an energy drink with anti-fatigue function.

[0013] An energy drink with anti-fatigue function, comprising the following raw materials by mass fraction: 75-90 parts of drinking water, 5-8 parts of oyster peptide, 1-3 parts of natural taurine, 2-5 parts of inositol, 3-8 parts of nicotinamide, 0.02-0.05 parts of vitamin B6, and 1-2 parts of vitamin B6. 12 The oyster peptide is 0.03-0.06 parts, dietary fiber is 1-3 parts, sweetener is 5-20 parts, and acidity regulator is 0.1-0.3 parts; the preparation method of the oyster peptide is as described above.

[0014] The advantages and beneficial effects of the present invention are:

[0015] The present invention prepares a protein hydrolyzate with more bitter amino acids exposed and less umami amino acids exposed by screening proteases, determining the cleavage sites and the end point of enzymolysis, and combines membrane separation technology to obtain oyster peptide powder with high purity, good flavor, low fat content and no fishy smell in the later stage, which is conducive to a wider application range of oyster peptides as food raw materials. The present invention optimizes the preparation process of oyster peptide powder, obtains peptide powder with better flavor and better stability in beverage applications, and solves the problem of fishy precipitation of oyster peptide powder in beverage product applications in the current market.

[0016] The present invention uses raw materials rich in natural taurine, scientific formulas and relatively simple production processes to solve the problems of existing energy drinks, such as high sugar content, caffeine dependence, and failure to take human health into consideration; the present invention uses pure natural extracted oyster peptides to effectively relieve human fatigue, improve nutrient digestion and absorption and energy utilization, and relieve sub-health conditions such as mental depression. The energy drink formula with anti-fatigue function provided by the present invention has been certified as low GI by a third-party testing agency, and its GI value is 30 to 40 (≤55) through preliminary experimental evaluation, and can be widely used in many types of people, such as students, the elderly, people who lose weight and people with type 2 diabetes. DETAILED DESCRIPTION

[0017] The technical solution of the present invention is further described below in conjunction with specific embodiments.

[0018] When developing oyster peptide beverages, in addition to ensuring their functionality and digestibility, the preparation process must also be improved to avoid problems such as poor flavor and unstable system during the shelf life of the beverage.

[0019] The basic flavors of peptides are sour, sweet, bitter, salty, and fresh. For the same raw material, the specificity of different enzymes and their shearing sites have great differences in the molecular weight and amino acid composition of the enzymatic hydrolysate, and this difference will be accompanied by strong differences in sensory perception. Oyster peptides are delicious and contain rich flavor amino acids (glutamic acid, aspartic acid, glycine, and alanine). Studies have found that the proportion of fresh and sweet amino acids in its enzymatic hydrolysate is as high as 48.1%, and the proportion of bitter amino acids is as high as 51.9%. The oyster peptide powders on the market are all exposures of some small umami peptides, and in actual preparation, the rich umami taste is inversely proportional to the flavor of the beverage. In addition, in the traditional beverage sterilization process, high temperature and high pressure or pH <4 will cause the denaturation of proteins and free fatty acids in the raw materials and auxiliary materials, and it is difficult to maintain good flavor and stability during the shelf life.

[0020] Embodiment 1:

[0021] This example studies the enzyme preparation and conditions for enzymolysis by experimental analysis.

[0022] 1. Enzyme preparation screening: By screening five kinds of enzymes, namely neutral protease, alkaline protease, aquatic protease, flavor protease, and animal compound enzyme, different combinations were made, the end point of enzymolysis was determined by the residue content, and the enzyme was inactivated to obtain the crude extract of oyster peptides for sensory evaluation. Referring to the national standard GB / T 16291.1-2012, the sensory evaluation team of this study selected 18 men and women without taste disorders, including 10 women and 8 men (aged between 25 and 32), and the temperature of the tasting room was controlled at room temperature (25±2℃). According to the characteristics of the experiment, the reference solution was used to train the team members. The reference solutions are as follows: umami (sodium glutamate solution, 15g / L); sweetness (sucrose solution 16g / L); sourness (citric acid solution 1g / L); astringency (quercetin solution 0.5g / L); bitterness (caffeine 0.5g / L).

[0023] The taste of characteristic flavor is relatively complex. This evaluation uses oyster soup made from oyster water boiled in plain water as the reference solution (10g / L boiled for 20min). The flavor score is 10 points based on the six reference solutions, and the relative flavor of the enzymatic hydrolysate is evaluated. And the score is judged from 0-10, and the higher the score, the heavier the flavor. The evaluation is divided into two steps: smelling and tasting. The oyster extract is kept at a constant temperature of 35°C and stored in a 20mL disposable plastic cup. The subjects need to smell its flavor first, and then taste it back and forth on the front, middle, back and sides of the tongue for about 15S, spit it out or swallow it, rinse the mouth with distilled water, and repeat the previous operation after 5min. The final result is the average of the scores of each group member.

[0024] Table 1 Sensory evaluation of different oyster hydrolysates

[0025] variety Umami sour bitterness Astringency Sweetness Characteristic flavor Neutral protease 5 4 8 6 3 4 Alkaline protease 7 6 7 4 4 7 Aquatic protease 9 7 4 8 4 8 Flavor protease 7 7 3 5 6 9 Animal compound protease 5 5 6 5 3 4

[0026] Note: All ratings remove the highest and lowest scores and take the average value

[0027] According to Table 1, the order of freshness from high to low is: aquatic protease > flavor protease = alkaline protease > animal complex protease (trypsin: pepsin: alkaline protease = 5.5:3.5:1) = neutral protease.

[0028] The order of bitterness from high to low is: neutral protease > alkaline protease > animal compound protease > aquatic protease > flavor protease

[0029] The characteristic flavors are ranked from high to low as follows: flavor protease > aquatic protease > alkaline protease > neutral protease = animal complex protease.

[0030] The purpose of this preparation is to obtain an oyster hydrolysate with a bitter main taste and a lighter umami taste and characteristic flavor, so the neutral protease and animal compound protease in Table 1 were comprehensively selected.

[0031] Embodiment 2:

[0032] The oysters are manually shelled and the edible parts are taken out and frozen to make frozen plates, and then homogenized, 0.05-0.3% animal composite protease (including 0.025%-0.15wt% trypsin, 0.015%-0.1wt% pepsin, and 0.01%-0.05wt% alkaline protease) is added, and enzymolysis is performed for 1-7h, and the enzyme is inactivated at 95°C to obtain a crude oyster peptide extract; activated carbon and diatomaceous earth are added to the crude peptide extract, and the mixture is stirred at a temperature of 50-80°C for 15-20min, and then plate and frame filtration is performed, and the filtered filtrate is desalted by using a membrane separation technology, and the desalted solution is spray-dried to obtain an oyster peptide raw material.

[0033] Embodiment 3:

[0034] The oysters are manually shelled and the edible parts are taken out and frozen to make frozen plates, and then the plates are homogenized, 0.01-0.2% neutral protease is added, and the enzyme is hydrolyzed for 1-7 hours, and the enzyme is inactivated at 95°C to obtain a crude oyster peptide extract; activated carbon and diatomaceous earth are added to the crude peptide extract, and the mixture is stirred at 50-80°C for 15-20 minutes, and then plate and frame filtration is performed, and the filtrate after filtration is desalted by using a membrane separation technology, and the desalted solution is spray-dried to obtain an oyster peptide raw material.

[0035] Embodiment 4:

[0036] The oysters are manually shelled and the edible parts are taken out and frozen to make frozen plates, and then homogenized, 0.05-0.3% animal composite protease (trypsin 0.025%-0.15wt%, pepsin 0.015%-0.1wt%, alkaline protease 0.01%-0.05wt%) and 0.01-0.2% neutral protease are added, and enzymolysis is performed for 1-7h, and the enzyme is inactivated at 95°C to obtain a crude oyster peptide extract; activated carbon and diatomaceous earth are added to the crude peptide extract, and the temperature is 50-80°C, and stirred for 15-20min, and then plate and frame filtration is performed, and the filtered filtrate is desalted by membrane separation technology, and the desalted solution is spray-dried to obtain an oyster peptide raw material.

[0037] The degree of hydrolysis of the selected enzymes was tested using the degree of protein hydrolysis as the evaluation condition: Relevant experimental data showed that the degree of protein hydrolysis was proportional to the enzymatic hydrolysis time, but the stability of the enzymatic hydrolysate showed a decreasing trend after the enzymatic hydrolysis time reached a certain value, so it was crucial to evaluate the impact of the degree of protein hydrolysis on the product. The nitrogen content of the oyster liquid raw material and the enzymatic hydrolysate was determined by the Kjeldahl method, and the ammonia nitrogen content was determined by the formaldehyde titration method. The calculation formulas for protein recovery and protein hydrolysis degree are as follows:

[0038] Degree of protein hydrolysis (DH) = (F t -F0) / N×100%

[0039] F t: The content of free amino nitrogen in oyster hydrolysate (g / 100mL);

[0040] F0: content of free amino nitrogen in oyster liquid (before enzymatic hydrolysis) (g / 100mL);

[0041] N: protein nitrogen content in oyster liquid (before enzymatic hydrolysis) (g / 100mL).

[0042] Table 2 Effects of different enzyme preparations and enzymatic hydrolysis time on protein hydrolysis degree

[0043]

[0044] As shown in Table 2, the degree of protein hydrolysis of the three samples is Example 4> Example 2> Example 3.

[0045] The oyster peptide obtained by the preparation method of the above embodiment 4 has a solubility of up to 99%. After analysis, the essential amino acid composition of oyster peptide reaches 55.7g / 100g, which is higher than milk protein (50.0g / 100g) and egg protein (49.7g / 100g), the polysaccharide content is ≥15.0%, and the natural taurine content is ≥1500mg / 100g; good stability, coexistence stability with other components, good storage stability; low viscosity; good processability; low osmotic pressure and low allergy; and good palatability. That is, the oyster peptide prepared in this embodiment 4 is suitable for the development of oyster peptide energy drinks.

[0046] In addition, the oyster peptide added in Example 4 is preferably derived from triploid oysters from exclusive natural waters, and oysters from October to April of the following year are selected. They are pollution-free, large in size, meaty, and rich in nutrients. Of course, high-quality oyster peptide powder can also be obtained by selecting oysters from other production areas and months.

[0047] Embodiment 5:

[0048] An oyster peptide energy drink is prepared from the following ingredients: 90 parts of drinking water, 6 parts of oyster peptide powder (commercially available product), 3 parts of taurine, 2 parts of inositol, 4 parts of nicotinamide, 0.04 of B6, 12 0.05, 3 parts of dietary fiber, 20 parts of sweetener, 0.2 parts of acidity regulator, 0.03 parts of edible flavor, 0.04 parts of preservatives, 0.08 parts of lemon yellow, and 0.02 parts of allura red.

[0049] The preparation method of the oyster peptide energy drink comprises the following steps:

[0050] (1) Add the formula amount of water, 6 parts of oyster peptide, 3 parts of taurine, 2 parts of inositol, 4 parts of niacinamide, 0.04 B6, 120.05, 3 parts of dietary fiber, 20 parts of sweetener, 0.2 parts of acidity regulator, 0.8 parts of pectin, 0.3 parts of soybean polysaccharide, 0.03 parts of edible flavor, 0.04 parts of preservative, 0.03 parts of tartrazine, 0.07 parts of brilliant blue, put them into a mixing tank in sequence, stir for 3-5 minutes, add 0.2 parts of acidity regulator at room temperature and mix well;

[0051] (2) Filtering through a filter with a filtration level of 10 to 50 μm, the filtered liquid is transparent, odorless, and has no visible matter;

[0052] (3) Fill the clarified liquid, sterilize the filled product and then seal it.

[0053] Embodiment 6:

[0054] An oyster peptide energy drink is prepared from the following raw materials: 90 parts of drinking water, 6 parts of oyster peptide powder (prepared according to Example 4), 3 parts of taurine, 2 parts of inositol, 4 parts of nicotinamide, 0.04 B6, 12 0.05, 3 parts of dietary fiber, 20 parts of sweetener, 0.2 parts of acidity regulator, 0.03 parts of edible flavor, 0.04 parts of preservatives, 0.08 parts of lemon yellow, and 0.02 parts of allura red.

[0055] The preparation method of the oyster peptide energy drink comprises the following steps:

[0056] (1) Add the formula amount of water, 6 parts of oyster peptide, 3 parts of taurine, 2 parts of inositol, 4 parts of niacinamide, 0.04 B6, 12 0.05, 3 parts of dietary fiber, 20 parts of sweetener, 0.2 parts of acidity regulator, 0.8 parts of pectin, 0.3 parts of soybean polysaccharide, 0.03 parts of edible flavor, 0.04 parts of preservative, 0.03 parts of tartrazine, 0.07 parts of brilliant blue, put them into a mixing tank in sequence, stir for 3-5 minutes, add 0.2 parts of acidity regulator at room temperature and mix well;

[0057] (2) Filtering through a filter with a filtration level of 10 to 50 μm, the filtered liquid is transparent, odorless, and has no visible matter;

[0058] (3) Fill the clarified liquid, sterilize the filled product and then seal it.

[0059] Evaluation experiments were performed on Examples 5 and 6:

[0060] 1) Paired comparison method to evaluate the sensory properties of oyster peptide beverages

[0061] Referring to the national standard GB / T 16291.1-2012, the sensory evaluation panel of this study selected 16 males and females without taste disorders, including 10 females and 6 males (aged between 25 and 32), and the temperature of the tasting room was controlled at room temperature (25±2°C). The beverages prepared in Example 5 and Example 6 were placed at 4°C, 25°C and 37°C for 3 months, respectively. The appearance and quantity of the samples were consistent, and each taster was given a group of samples. The aroma, sweetness, sourness, fishy smell (whether it is anti-fishy) and freshness of the samples were evaluated for differences, and the results were filled in the evaluation table of Table 3.

[0062] Table 3 Directed pairwise comparison evaluation table

[0063]

[0064] The statistical results of the differences between the 2 oyster peptide drinks by 16 tasters are as follows:

[0065] Table 4 Statistical table of difference pair evaluation results

[0066]

[0067]

[0068] From the above table, it can be seen that only 1 of the 16 people said that the difference was not obvious, and the rest said that Example 1 had a serious anti-smell; in terms of aroma index, 13 people indicated that Example 6 had a better aroma, and 1 person said there was no difference; in terms of sour and sweet taste, since the addition of sour and sweeteners in Examples 5 and 6 was the same, 7 people said that the difference was not obvious, and 7 people said that Example 6 had a better sour and sweet taste; in terms of freshness, 14 people said that Example 6 had a better freshness, and 1 person said that the difference was not obvious. This shows that the homemade oyster peptide powder has a better flavor during the shelf life.

[0069] To determine the effect of oyster peptide powder on the stability of beverages, the beverages in Example 5 and Example 6 were placed at 4°C, 25°C and 37°C for 3 months to observe their product stability.

[0070] Table 5 Effects of different enzyme preparations and enzymatic hydrolysis time on the system

[0071]

[0072]

[0073] At 4°C, the beverage prepared from the collected oysters in Example 5 began to precipitate on the third day of the first week, at 25°C, precipitate began to form from the fourth week, and at 37°C, precipitate began to form from the third week. However, there was no precipitation in Example 6 at elevated temperatures and for a long time, indicating that the oyster peptide added in Example 6 made the overall system more stable.

[0074] The oyster peptide powder prepared by the enzymatic hydrolysis method provided in Example 6 has a more delicate and coordinated taste and a more stable and excellent flavor in beverage preparation applications. Compared with oyster peptide products on the market, beverages prepared by this method have a better taste, a more stable system, no anti-fishy smell during the shelf life, and a better physical sensation for consumers.

[0075] The above description shows that the peptide powder prepared in Example 4 not only has a stable flavor and no fishy smell, but also does not produce precipitation under the conditions of high temperature and high pressure (sterilization) or pH < 4 (acidity regulator).

[0076] 2. Animal experiment verification on relieving physical fatigue:

[0077] 1 Materials and Methods

[0078] 1.1 Sample: The oyster peptide energy drink prepared in Example 6 (powder formula without water) was sent for inspection. The sample was a milky white solid. It was prepared with ultrapure water to a corresponding concentration before use. The approved net content of the sample was 400 g / bag. The recommended daily intake of oyster peptide energy drink per person was 330 ml / 60 kg·BW. Storage conditions: 4°C. Solvent: ultrapure water.

[0079] 1.2 Comparative sample: Red Bull Vitamin Flavored Drink, produced by Tianshi Pharmaceutical Healthcare Co., Ltd. (Thailand), the sample is a yellow liquid, and the recommended daily intake per person is 250 ml. Sample batch number: 20221205, shelf life: 12 months. Before use, the vitamin flavored drink was rotary evaporated at low temperature (50°C) into 1000 ml for standby use.

[0080] 1.2 Experimental animals: 168 SPF ICR mice; male; initial weight range: 19-23g

[0081] 1.3 Dose selection and administration of test substances: A negative control group (ultrapure water) was set up, three dose groups of oyster peptide energy drink: low dose group (14.0ml / kg·BW), medium dose group (46.7ml / kg·BW) and high dose group (140.0ml / kg·BW) (equivalent to 3 times, 10 times and 30 times the daily intake of humans, respectively), and three dose groups of Red Bull vitamin flavored drink: low dose (12.5ml / kg·BW), medium dose (41.7ml / kg·BW), high dose group (125ml / kg·BW) (equivalent to 3 times, 10 times and 30 times the daily intake of humans, respectively). Administration of test substances and comparative samples: oral gavage, 0.2ml / 10g·BW.

[0082] 1.4 Main instruments and reagents: swimming box (50cm×50cm×40cm), SPN3001F electronic balance, lead scale, stopwatch, Hitachi 7020 fully automatic biochemical analyzer, UV2100 ultraviolet spectrophotometer, centrifuge, constant temperature water bath; lactate meter (Lactate SCOUT), lactate test strips (Lactate Scout Sensors Big Pack 72+, batch number: 1909827254), liver glycogen / muscle glycogen detection kit (BB-4723, Shanghai Beibo Biotechnology Co., Ltd.), urea nitrogen kit (MAKER biochemical kit).

[0083] 1.5 Experimental method: Grouping: 280 animals were divided into 4 subgroups, with 70 animals in each subgroup. The animals in each subgroup were randomly divided into 7 groups according to body weight, namely, negative control group, oyster peptide energy drink low-dose group, medium-dose group, high-dose group and Red Bull vitamin flavored drink low-dose group, medium-dose group, and high-dose group, with 10 animals in each group. Weigh a certain amount of oyster peptide energy drink concentrated solid powder and dissolve it in ultrapure water to prepare a solution of the required concentration. The Red Bull vitamin flavored drink was rotary evaporated at low temperature (50°C) to 2000ml, which was used as the gavage solution of the required concentration for the high-dose group, and then diluted with ultrapure water to prepare a solution of the corresponding concentration. Animals in each group were given different doses of oyster peptide energy drink and Red Bull vitamin flavored drink by gavage, and the negative control group was given ultrapure water. The gavage capacity of mice was 0.2ml / 10g·BW. Gavage once a day at the same time. Weigh and adjust the gavage volume weekly, and gavage for 30 consecutive days. Mice in each group were fed with ordinary feed and had free access to food and water. After 30 days, the physical fatigue relief function of mice in each subgroup was tested.

[0084] 1.5.1 Weighted swimming experiment: 30 minutes after the last administration of the test sample to 10 mice in each dose group of subgroup 1, 5% of the body weight of lead was loaded on the base of the mouse's tail. The mice were then placed in a swimming box (water depth ≥30 cm, water temperature 25℃±1.0℃) to swim. The time from the start of swimming to the death of the mice was recorded as the weighted swimming time of the mice.

[0085] 1.5.2 Serum urea nitrogen determination: 30 minutes after the last administration of the test substance to the 10 mice in each dose group of the second subgroup, they swam without weight in water at 30°C for 90 minutes. After resting for 60 minutes, blood was collected and serum was separated. Serum urea nitrogen was determined using a Hitachi 7020 fully automatic biochemical analyzer, and the reagent used was a MAKER biochemical kit.

[0086] 1.5.3 Determination of liver glycogen: 30 minutes after the last administration of the test substance to 10 mice in each dose group of subgroup 3, the animals were killed, the livers were removed, rinsed with physiological saline and dried with filter paper, 0.1-0.2 g of the livers were accurately weighed, and the test was performed according to the instructions of the kit.

[0087] 1.5.4 Lactic acid determination: 30 minutes after the last intragastric administration of 10 mice in each dose group of the fourth subgroup, 1-2 mm of tail blood was cut, and the first drop of blood was wiped off with a cotton ball. A trace amount of tail blood was collected and the blood lactate content of the mice in the resting state was measured by a lactate meter. Then the mice swam in 30°C water without weight for 10 minutes. The blood lactate value of the mice immediately after exercise and after rest was measured by a lactate meter at 0 and 20 minutes after swimming.

[0088] 1.6 Experimental data statistics: The variance analysis method was used to compare the differences in animal body weight, urea nitrogen, liver glycogen, blood lactate (before swimming, 0 min after swimming, 20 min after swimming) between each experimental group and the negative control group, p < 0.05 was considered statistically significant; due to unequal variances, the weighted swimming time and the area under the blood lactate curve were transformed by inverse conversion and COS conversion respectively, and the differences between each experimental group and the negative control group were compared by variance analysis, p < 0.05 was considered statistically significant.

[0089] 1.7 Result determination: Weighted swimming time: If the weighted swimming time of the test sample group is significantly longer than that of the control group, and the difference is significant, the experimental result can be determined to be positive; Urea nitrogen: If the serum urea of ​​the test sample group is lower than that of the negative control group, and the difference is significant, the experimental result can be determined to be positive; Liver glycogen: If the liver glycogen content of the test sample group is significantly higher than that of the negative control group, and the difference is significant, the experimental result can be determined to be positive; Blood lactate: The area under the blood lactate curve at three time points is used as the judgment standard. If the area of ​​any test group is smaller than that of the negative control group, and the difference is significant, the experimental result can be determined to be positive. If the weighted swimming test result is positive, and any two of the three biochemical indicators of blood lactate, serum urea, and liver glycogen are positive, it can be determined that the test sample has the function of relieving physical fatigue.

[0090] 2 Results:

[0091] 2.1 Animal weight gain

[0092] Table 6 shows that the high-dose group of the second subgroup and the fourth subgroup B had a significant effect (p < 0.05) during the feeding of oyster peptide energy drinks, and there was no significant difference between the other components and the negative control. Therefore, it is considered that the oral administration of oyster peptide energy drinks and Red Bull vitamin drinks is not the main factor affecting the significant increase in mouse weight, and there is no dose-response relationship between the two. In summary, oyster peptide energy drinks and Red Bull vitamin drinks had no effect on mouse weight in the 30-day oral administration experiment.

[0093] Table 6 Effects of oyster peptide energy drink on body weight gain in mice (g, )

[0094]

[0095]

[0096] Note: *. Indicates significant difference compared with the negative control group (p<0.05);

[0097] **. Indicates that there is a significant difference compared with the negative control group (p<0.01).

[0098] A. Oyster peptide energy drink

[0099] B. Red Bull Vitamin Flavored Drink

[0100] Table 7 Effect of oyster peptide energy drink on weight-bearing swimming time of mice

[0101]

[0102] Note: ** indicates significant difference compared with the negative control group (p<0.01)

[0103] Animal experiments have shown that after oral administration to mice for 30 consecutive days, the weighted swimming time of the low-dose oyster peptide energy drink group and the medium-dose Red Bull vitamin flavored drink group was significantly higher than that of the negative control group (p < 0.01), and the weighted swimming time of the low-dose oyster peptide energy drink group was 8.65 ± 3.75 min, which was higher than that of the medium-dose Red Bull vitamin energy drink group (7.66 ± 3.16 min). The weighted swimming time of mice was significantly prolonged.

[0104] Table 8 Effects of oyster peptide energy drink on serum urea nitrogen and liver glycogen in mice

[0105]

[0106]

[0107] Note: ** .Indicates that there is a significant difference compared with the negative control group (p<0.01).

[0108] After 30 days of oral administration of oyster peptide energy drink and Red Bull vitamin drink, compared with the negative control group, the serum urea nitrogen level of mice in the high-dose group was significantly reduced after swimming for 90 minutes without weight bearing and resting for 60 minutes (p<0.01); the serum urea nitrogen of the high-dose oyster peptide energy drink group was 7.3±1.3mmol / L, and that of the Red Bull vitamin flavored drink was 7.9±2.2mmol / L (see Table 8).

[0109] Effect of oyster peptide energy drink on liver glycogen: After being given oyster peptide energy drink for 30 days, there was no significant change in the liver glycogen level of mice in each dose group compared with the negative control group; after being given Red Bull vitamin flavored drink for 30 days, the liver glycogen level of mice in the medium dose group and high dose group was significantly reduced compared with the negative control group, and the difference was significant (p<0.01) (see Table 8).

[0110] Table 9 Effect of oyster peptide energy drink on blood lactate in mice (mmol / L)

[0111]

[0112] Note: * .Indicates that there is a significant difference compared with the negative control group (p<0.05);

[0113] ** .Indicates that there is a significant difference compared with the negative control group (p<0.01).

[0114] a .After logarithmic transformation, the data were normally distributed, the variances of each group were equal, and one-way analysis of variance was used;

[0115] b After logarithmic transformation, the data were considered to be non-normally distributed and rank sum analysis was performed.

[0116] Animal experiments have shown that after mice were given 14.0, 46.7, and 140.0 ml / kg·BW oyster peptide energy drinks by gavage for 30 consecutive days, the mice's weighted swimming time was significantly prolonged; the serum urea nitrogen level was significantly reduced; the area under the blood lactate curve at three time points before swimming, 0 min after swimming, and 20 min after swimming was significantly reduced; but there was no significant effect on the liver glycogen level of mice at rest. According to the judgment standard for relieving physical fatigue in the Ministry of Health's "Technical Specifications for Inspection and Evaluation of Health Foods" (2003 edition), it can be considered that marine active peptide energy drinks have the function of relieving physical fatigue.

[0117] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any technician familiar with the profession can use the above technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. A method for preparing oyster peptide, characterized in that: The method comprises the following steps: enzymolyzing the edible part of the oyster, and the selected enzymes are animal composite protease and neutral protease; the animal composite protease comprises trypsin, pepsin and alkaline protease.

2. The preparation method according to claim 1, characterized in that Specifically, the edible parts of fresh oysters are frozen and homogenized, 0.05%~0.3wt% animal composite protease and 0.01%~0.2wt% neutral protease are added, and enzymolysis is performed for 1~7h to obtain a crude oyster peptide extract; the crude peptide extract is decolorized and then filtered, the filtered filtrate is desalted, and the desalted solution is spray-dried to obtain oyster peptides; the 0.05%~0.3wt% animal composite protease includes 0.025%~0.15 wt% trypsin, 0.015%~0.1 wt% pepsin, and 0.01%~0.05 wt% alkaline protease.

3. The preparation method according to claim 2, characterized in that: The desalination adopts membrane separation technology.

4. Use of the oyster peptide prepared according to claim 1 in beverage products.

5. Use of the oyster peptide prepared according to claim 1 in an energy drink with anti-fatigue function.

6. An energy drink with anti-fatigue function, characterized in that: The invention comprises the following raw materials by mass fraction: 75-90 parts of drinking water, 5-8 parts of oyster peptide prepared according to claim 1, 1-3 parts of natural taurine, 2-5 parts of inositol, 3-8 parts of nicotinamide, 0.02-0.05 parts of vitamin B6, and 1-2 parts of oyster peptide prepared according to claim 1. 12 0.03-0.06 parts, dietary fiber 1-3 parts.

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