CN117778505A - Preparation method and application of oyster peptide - Google Patents
Preparation method and application of oyster peptide Download PDFInfo
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- CN117778505A CN117778505A CN202311802830.1A CN202311802830A CN117778505A CN 117778505 A CN117778505 A CN 117778505A CN 202311802830 A CN202311802830 A CN 202311802830A CN 117778505 A CN117778505 A CN 117778505A
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- oyster peptide
- oyster
- parts
- peptide
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- Non-Alcoholic Beverages (AREA)
Abstract
The invention discloses a preparation method and application of oyster peptide, belonging 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 and homogenizing fresh oyster edible parts, adding 0.05-0.3 wt% of animal compound protease and 0.01-0.2 wt% of neutral protease, and performing enzymolysis for 1-7 h to obtain oyster peptide crude extract; decolorizing the crude oyster peptide extract, filtering, desalting the filtrate, and spray drying to obtain oyster peptide powder. The preparation method optimizes the preparation process of the oyster peptide, obtains the peptide powder with better flavor and better stability in beverage application, and solves the problem of fishy smell prevention and precipitation of the oyster peptide in beverage product application in the current market.
Description
Technical Field
The invention belongs to the technical field of marine bioactive substances, and particularly relates to a preparation method and application of oyster peptide.
Background
With the progress and development of society, the people stay up and work overtime, and the mental stress is increased continuously, so that the people have serious overdraft of energy and physical strength, and the human body system is overloaded to operate, fatigued, weak and weak, and the work and life are seriously affected. In general, for this sub-healthy state without organic lesions, most people will selectively ignore it, burying the risk for the eventual development of the disease state. Therefore, the health care tea can regulate the body function, get rid of sub-health state, and control the occurrence and development of diseases from the source.
At present, most of energy drinks on the market contain taurine, caffeine, guarana, white granulated sugar, vitamins, minerals and the like, and the energy drinks have good refreshing effect on mature working population and high-strength operation fatigue-prone population, but are weak in caffeine metabolism for student population with incomplete physical development, and a small amount of caffeine is likely to reach toxic dosage to cause harm to the students. In addition, taurine in the formula is basically an artificial synthetic raw material, and does not have the biological activity of natural taurine. Research shows that taurine is mainly metabolized by kidneys after being absorbed by human bodies, so that the balance of the taurine content in the bodies is maintained, and excessive intake tends to cause kidney burden. Frequent consumption of such beverages in large quantities is prone to dependency and has relatively obvious side effects. Meanwhile, a large amount of added white granulated sugar is not suitable for people with weight loss, blood sugar regulation dysfunction and type 2 diabetes. Therefore, the market is urgent to provide a novel energy beverage which can fundamentally solve the pain points for consumers, can meet the market trend, meets the demands of the consumers, has natural and safe raw material components, and has the functions of resisting fatigue, resisting oxidation, supplementing energy, refreshing, soothing the nerves and the like.
However, the energy peptide beverage products currently on the market generally exhibit very severe anti-fishy and cloudy conditions, and the products themselves have poor flavor, especially after 8 months of shelf life, often exhibit macroscopic significant flocculation and intolerable characteristic flavors and fishy odors.
Disclosure of Invention
The invention aims to provide a preparation method and application of oyster peptide, which are used for overcoming the defects of the prior art.
The invention achieves the aim by the following technical scheme:
a process for preparing oyster peptide includes such steps as enzymolysis of edible oyster part, and the chosen enzymes are animal composite proteinase and neutral proteinase.
Further, the preparation method specifically comprises the following steps: freezing and homogenizing fresh oyster edible parts, adding 0.05-0.3 wt% of animal compound protease and 0.01-0.2 wt% of neutral protease, and performing enzymolysis for 1-7 h to obtain oyster peptide crude extract; and (3) decoloring the peptide crude extract, filtering, desalting the filtered filtrate, and spray-drying the desalted solution to obtain oyster peptide.
Further, the desalination is by membrane separation technology.
The application of the oyster peptide in the preparation of beverage products.
The application of the oyster peptide in the preparation of the energy beverage with the anti-fatigue function.
An energy drink with an anti-fatigue function comprises the following raw materials in percentage by mass: 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 and vitamin B 6 0.02-0.05 part of vitamin B 12 0.03-0.06 part, 1-3 parts of dietary fiber, 5-20 parts of sweetener and 0.1-0.3 part of acidity regulator; the preparation method of the oyster peptide is as described above.
The invention has the advantages and beneficial effects that:
the invention prepares the protein hydrolysate with more bitter amino acid exposure and less fresh amino acid exposure through screening protease and judging enzyme cutting sites and enzymolysis end points, and the oyster peptide powder with high purity, good flavor, low fat content and no fishy smell in the later period can be obtained by combining a membrane separation technology, thereby being beneficial to the application range of oyster peptide serving as a food raw material. The preparation process of the oyster peptide powder is optimized, the peptide powder with better flavor and better stability in beverage application is obtained, and the problem of anti-fishy precipitation of the oyster peptide powder in beverage product application in the current market is solved.
The invention adopts a scientific formula and a relatively simple production process to solve the problems of high sugar content, caffeine dependence, no human health point of view and the like of the existing energy beverage by adopting the raw materials rich in natural taurine; the oyster peptide is extracted by pure natural method, so that the fatigue state of a human body is effectively relieved, the digestion and absorption of nutrient substances and the energy utilization are improved, and the sub-health states such as listlessness and the like are relieved. The energy drink formula with the anti-fatigue function provided by the invention is subjected to low GI authentication by a third party detection mechanism, and the GI value of the energy drink formula is estimated to be 30-40 (less than or equal to 55) through preliminary experiments, so that the energy drink formula can be widely applied to various crowds, such as students, old people, people with weight loss, type 2 diabetes mellitus and the like.
Detailed Description
The technical scheme of the invention is further described and illustrated below by combining with the embodiment.
When oyster peptide beverage is developed, the preparation process is improved in addition to ensuring the functionality and digestibility absorption of oyster peptide beverage so as to avoid the problem of unstable flavor system in the shelf life of the beverage.
The basic taste of peptide is expressed by five kinds of sour, sweet, bitter, salty and fresh, and the specificity of different enzymes and the shearing action sites of the different enzymes aiming at the same raw material have larger difference on the molecular weight and amino acid composition of enzymolysis products, and the difference is accompanied by strong difference of sense organs. The oyster peptide meat is delicious, and researches on the oyster peptide meat containing rich flavor-developing amino acids (glutamic acid, aspartic acid, glycine and alanine) find that the fresh sweet amino acid content in the enzymolysis liquid is up to 48.1 percent, the bitter amino acid content is up to 51.9 percent, and the oyster peptide powder on the market is exposed by some flavor-developing small peptides, and in actual blending, the flavor of the oyster peptide is rich and the flavor of the beverage is inversely proportional. In the traditional beverage sterilization process, the denaturation of protein and free fatty acid in raw and auxiliary materials can be caused by high temperature and high pressure or pH less than 4, and good flavor and stability are difficult to maintain in the shelf life.
Example 1:
the enzyme preparation selected for enzymolysis and the enzymolysis conditions are researched through experimental analysis.
1. Enzyme preparation screening: five enzymes of neutral protease, alkaline protease, aquatic protease, flavourzyme and animal compound enzyme are screened for different collocations, an enzymolysis end point is judged through the slag content, enzyme deactivation treatment is carried out, an oyster peptide crude extract is obtained for carrying out sensory evaluation, 18 men and women without dysgeusia are selected by a sensory evaluation group of the study according to national standard GB/T16291.1-2012, wherein 10 women 8 men (age between 25 and 32) are selected, and the temperature of a evaluating room is controlled at room temperature (25+/-2 ℃). The panelists were trained with the reference solution for the characteristics of the experiment. The reference solutions were as follows: umami taste (sodium glutamate solution, 15 g/L); sweet taste (sucrose solution 16 g/L); sour taste (citric acid solution 1 g/L); astringency (quercetin solution 0.5 g/L)); bitter taste (caffeine 0.5 g/L).
The taste of the characteristic flavor is complex, and oyster soup prepared by oyster water boiled with white water is used as a reference solution (10 g/L boiled for 20 min) for the evaluation. And (5) taking six reference liquids as reference flavors, wherein the flavor scores are 10 minutes, and evaluating the relative flavors of the enzymolysis liquids. And 0-10 points are used for judgment, and the higher the point is, the heavier the flavor is judged. The evaluation is divided into two steps of smelling and swallowing, oyster extract is kept at constant temperature to 35 ℃ and stored in a 20mL disposable plastic cup, a subject needs to smell the flavor of the oyster extract firstly, then the flavor of the oyster extract is tasted back and forth at the front, middle and rear sides of the tongue for about 15 seconds, the oyster extract can be spitted or swallowed, distilled water is used for rinsing, and the previous operation is repeated after 5 minutes. The final results were averaged over the panelist scores.
TABLE 1 sensory evaluation of different oyster enzymatic liquids
| Variety of species | Fresh flavor | Sour taste | Bitter taste | Astringency of the tea | Sweet taste | 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 |
| Flavoured protease | 7 | 7 | 3 | 5 | 6 | 9 |
| Animal composite protease | 5 | 5 | 6 | 5 | 3 | 4 |
Note that: all scores were removed from the highest score and the lowest score, and the remaining averages were taken
As can be seen from table 1, the fresh high to low sequences are: aqua protease > flavourzyme = alkaline protease > animal complex protease = neutral protease.
The bitter taste is sequentially from high to low: neutral protease > alkaline protease > animal complex protease > aquatic protease > flavor protease
The characteristic flavors are as follows in sequence from high to low: flavourzyme > aquaprotease > alkaline protease > neutral protease = animal complex protease.
The preparation aims at obtaining oyster enzymolysis liquid with main taste of bitter taste and weak flavor of fresh taste and characteristic taste, so the neutral protease and animal compound protease in table 1 are comprehensively selected.
Example 2:
manually removing shells of oysters, taking edible parts, freezing to prepare a frozen plate, homogenizing, adding 0.05-0.3% of animal compound protease, carrying out enzymolysis for 1-7 h, and inactivating enzyme at 95 ℃ to obtain oyster peptide crude extract; adding active carbon and diatomite into the peptide crude extract, stirring for 15-20min at 50-80 ℃, then filtering by a plate frame, desalting the filtered filtrate by a membrane separation technology, and performing spray drying treatment on the desalted solution to obtain oyster peptide raw materials.
Example 3:
manually removing shells of oysters, taking edible parts, freezing to prepare a frozen plate, homogenizing, adding 0.01-0.2% of neutral protease, carrying out enzymolysis for 1-7 h, and inactivating enzyme at 95 ℃ to obtain oyster peptide crude extract; adding active carbon and diatomite into the peptide crude extract, stirring for 15-20min at 50-80 ℃, then filtering by a plate frame, desalting the filtered filtrate by a membrane separation technology, and performing spray drying treatment on the desalted solution to obtain oyster peptide raw materials.
Example 4:
freezing the edible parts of oyster which are manually shelled to prepare a frozen plate, homogenizing, adding 0.05-0.3% of animal compound protease and 0.01-0.2% of neutral protease, carrying out enzymolysis for 1-7 h, and inactivating enzyme at 95 ℃ to obtain oyster peptide crude extract; adding active carbon and diatomite into the peptide crude extract, stirring for 15-20min at 50-80 ℃, then filtering by a plate frame, desalting the filtered filtrate by a membrane separation technology, and performing spray drying treatment on the desalted solution to obtain oyster peptide raw materials.
The selected enzymes were tested for their degree of hydrolysis using the degree of proteolysis as an evaluation condition: the related experimental data show that the proteolytic degree is in direct proportion to the enzymolysis time, but the stability of the enzymolysis liquid has a decreasing trend after the enzymolysis time reaches a certain value, so that the evaluation of the proteolytic degree is important to the influence of the product. And (3) determining the nitrogen content of the oyster liquid raw material and the enzymolysis liquid by using a Kjeldahl nitrogen determination method, and determining the ammonia nitrogen content by using a formaldehyde titration method. The calculation formulas of protein recovery and protein hydrolysis degree are as follows:
degree of proteolysis (DH) = (F) t -F 0 )/N×100％
F t The content of free amino nitrogen (g/100 mL) in oyster enzymolysis liquid;
F 0 free amino nitrogen content (g/100 mL) in oyster liquid (before enzymolysis);
n: protein nitrogen content (g/100 mL) in oyster liquid (before enzymolysis).
TABLE 2 influence of different enzyme preparations and enzymolysis time on the degree of proteolysis
As shown in Table 2, the degree of proteolysis of the three samples was in the order of example 4 > example 2 > example 3.
And the oyster peptide obtained by the preparation method of the example 4 has the solubility of up to 99 percent. According to analysis, the composition of essential amino acid of oyster peptide reaches 55.7g/100g, which is higher than milk protein (50.0 g/100 g) and egg protein (49.7 g/100 g), the polysaccharide content is more than or equal to 15.0%, and the natural taurine content is more than or equal to 1500mg/100g; good stability, coexistence stability with other components, good storage stability; low viscosity; good workability; low osmotic pressure and low allergy; has good palatability. That is, the oyster peptide prepared in example 4 was suitable for development of oyster peptide energy drink.
In addition, the oyster peptide added in example 4 is preferably derived from triploid oysters of the exclusive natural sea area, and oysters of 10 months to 4 months of the next year are selected, so that the oyster peptide powder has no pollution, big head, high meat quality and rich nutrition, and can be obtained from oysters of other producing areas and months.
Example 5:
an oyster peptide energy beverage is prepared from the following raw materials: 90 parts of drinking water, 6 parts of oyster peptide powder (commercial product), 3 parts of taurine, 2 parts of inositol, 4 parts of nicotinamide and B 6 Is 0.04, B 12 0.05 part of dietary fiber 3 parts, 20 parts of sweetener, 0.2 part of acidity regulator, 0.03 part of edible essence, 0.04 part of preservative, 0.08 part of lemon yellow and 0.02 part of allure red.
The preparation method of the oyster peptide energy beverage comprises the following steps:
(1) Mixing 6 parts of oyster peptide, 3 parts of taurine, 2 parts of inositol, 4 parts of nicotinamide and B 6 Is 0.04, B 12 0.05 part of dietary fiber 3 parts, sweetener 20 parts, acidity regulator 0.2 parts, pectin 0.8 parts, soybean polysaccharide 0.3 parts, edible essence 0.03 parts, preservative 0.04 parts, lemon yellow 0.03 parts and brilliant blue 0.07 parts are sequentially put into a stirring tank, stirred for 3-5 minutes, then acidity regulator 0.2 parts is added at normal temperature, and uniformly mixed;
(2) Filtering by a filter with the filtering grade of 10-50 mu m, wherein the filtered feed liquid is transparent, odorless and has no visual matters;
(3) And (5) filling the clear liquid, sterilizing the filled product, and sealing the cover.
Example 6:
an oyster peptide energy beverage 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 and B 6 Is 0.04, B 12 0.05 part of dietary fiber 3 parts, 20 parts of sweetener, 0.2 part of acidity regulator, 0.03 part of edible essence, 0.04 part of preservative, 0.08 part of lemon yellow and 0.02 part of allure red.
The preparation method of the oyster peptide energy beverage comprises the following steps:
(1) Mixing 6 parts of oyster peptide, 3 parts of taurine, 2 parts of inositol, 4 parts of nicotinamide and B 6 Is 0.04, B 12 0.05 part of dietary fiber 3 parts, sweetener 20 parts, acidity regulator 0.2 parts, and pectin0.8 part of soybean polysaccharide 0.3 part of edible essence 0.03 part of preservative 0.04 part of lemon yellow 0.03 part of brilliant blue 0.07 part of soybean polysaccharide, and sequentially placing the materials into a stirring tank, stirring for 3-5 minutes, adding 0.2 part of acidity regulator at normal temperature, and uniformly mixing;
(2) Filtering by a filter with the filtering grade of 10-50 mu m, wherein the filtered feed liquid is transparent, odorless and has no visual matters;
(3) And (5) filling the clear liquid, sterilizing the filled product, and sealing the cover.
Evaluation experiments were performed on examples 5 and 6:
1) Evaluation of oyster peptide beverage sensory Properties by paired comparison
With reference to national standard GB/T16291.1-2012, the sensory panel of the study selected 16 individuals for men and women without dysgeusia, of which 10 women 6 men (age between 25 and 32), and the temperature of the rated room was controlled at room temperature (25.+ -. 2 ℃). The beverages prepared in example 5 and example 6 were placed at 4 ℃,25 ℃ and 37 ℃ for 3 months, respectively, and the appearance and amount of the samples were identical, and each panelist was given a set of samples, respectively. The samples were evaluated for differences in aroma, sweetness, sourness, fishy smell (whether or not fishy smell is reversed), freshness, and the like, and the results are shown in the evaluation table of table 3.
Table 3 oriented pair comparison evaluation Table
The results of the difference between the 2 oyster peptide drinks by the 16 panelists were as follows:
TABLE 4 statistical table of differential pair-wise evaluation results
As can be seen from the above table, only 1 out of 16 people show insignificant differences, and the rest all show severe fishy smell in example 1; in terms of aroma index, 13 people indicate that the aroma of example 6 is better, and 1 person indicates no difference; in terms of sweet and sour sensation, 7 persons indicate no obvious difference and 7 persons indicate that the sweet and sour sensation of example 6 is better because the addition of the acid sweeteners of examples 5 and 6 is the same; the 14 persons in the freshness degree indicate that the freshness degree of example 6 is better, and the 1 person indicates that the difference is not obvious. Therefore, the self-made oyster peptide powder has better flavor in shelf life.
To determine the effect of oyster peptide powder on the stability of the beverage, the beverages of example 5 and example 6 were placed at 4 ℃,25 ℃ and 37 ℃ for 3 months, respectively, and the product stability was observed.
TABLE 5 influence of different enzyme preparations and enzymolysis times on the system
The beverage prepared by the external harvest of the finished oyster in example 5 at 4℃starts to precipitate on the third day of the first week, starts to precipitate on the fourth week at 25℃and starts to precipitate on the third week at 37 ℃. Whereas example 6 did not precipitate at both elevated temperature and for a long period of time, it was demonstrated that the oyster peptide added in example 6 made the whole system more stable.
The oyster peptide powder prepared by the enzymolysis method provided in the example 6 has finer and more consistent taste and more stable and excellent flavor in beverage preparation application. Compared with oyster peptide products on the market, the beverage prepared by the method has better taste, more stable system, no fishy smell in shelf life and better body feeling of consumers.
The above demonstrates that the peptide powder prepared in example 4 is not only stable in flavor and free of fishy smell, but also does not precipitate under high temperature and high pressure (sterilization) or pH < 4 (acidity regulator).
2. Animal experiments for relieving physical fatigue prove that:
1 materials and methods
1.1 sample: the oyster peptide energy beverage (powder formulation without water) prepared in example 6 was submitted to inspection, the sample submitted to inspection was a milky white solid, prepared with ultrapure water to the corresponding concentration immediately before use, the sample approved net content: 400 g/bag, oyster peptide energy beverage recommended daily intake per person is 330ml/60 kg.BW, preservation conditions: preserving at 4 ℃. Solvent: ultrapure water.
1.2 comparative samples: the red ox vitamin flavor beverage is produced by Tiansi medical health care Co., ltd (Thailand) and is yellow in color, and the daily intake of 250ml per person is recommended. Sample lot number: 20221205, shelf life: for 12 months. The vitamin flavor beverage is rotationally evaporated to 1000ml at low temperature (50 ℃) for standby before use.
1.2 experimental animals: 168 SPF-class ICR mice; a male; initial body weight range: 19-23g
1.3 dose selection and mode of administration of the test substance: a negative control group (ultrapure water) was set, oyster peptide energy drink three dose groups: low dose group (14.0 ml/kg-BW), medium dose group (46.7 ml/kg-BW) and high dose group (140.0 ml/kg-BW) (corresponding to 3, 10 and 30 times the daily intended intake of humans, respectively), three dose groups of the vitamin-in-red beverage: low dose (12.5 ml/kg.BW), medium dose (41.7 ml/kg.BW), high dose group (125 ml/kg.BW) (corresponding to 3 times, 10 times and 30 times daily intake of human respectively). Test substance and comparative sample administration mode: the stomach is irrigated by mouth, 0.2ml/10 g.BW.
1.4 major instrumentation and reagents: swimming case (50 cm. Times.50 cm. Times.40 cm), SPN3001F electronic balance, lead skin, stopwatch, hitachi 7020 full-automatic biochemical analyzer, UV2100 ultraviolet spectrophotometer, centrifuge, constant temperature water bath; lactic acid analyzer (Lactate SCOUT), lactic acid test paper (Lactate Scout Sensors Big Pack 72+, lot number: 1909827254), hepatic glycogen/myoglycogen detection kit (BB-4723, shanghai Bei Bo Biotech Co., ltd.), urea nitrogen kit (MAKER Biochemical kit).
1.5 test method: grouping: 280 animals were divided into 4 subgroups of 70 animals each. Animals in each subgroup were randomly divided into 7 groups, namely a negative control group, an oyster peptide energy drink low dose group, a medium dose group, a high dose group, and a red cow vitamin flavor drink low dose group, a medium dose group, a high dose group, each group of 10 animals. Weighing a certain amount of oyster peptide energy beverage concentrated solid powder, dissolving with ultrapure water, and preparing into solution with required concentration. The red ox vitamin beverage is rotary evaporated to 2000ml at low temperature (50 deg.c) to form the high dosage group gastric lavage solution of required concentration, and the solution is diluted with ultrapure water to prepare solution of corresponding concentration. Each group of animals was respectively given different doses of oyster peptide energy drink and red cow vitamin flavor drink for lavage, the negative control group was given ultrapure water, and the capacity of the mice for lavage was 0.2ml/10 g.BW. The stomach was irrigated 1 time a day at the same time. The amount of the stomach is weighed and adjusted every week, and the stomach is continuously irrigated for 30 days. Each group of mice was fed normal feed and had free access to food and water. After 30 days, each subgroup of mice was tested for physical fatigue relieving function.
1.5.1 load swimming experiments: after 10 mice in each dose group of subgroup 1 were last given the test sample for 30min, the tail root was loaded with 5% weight of lead skin, then the mice were placed in a swimming box for swimming (water depth: 30cm, water temperature: 25 ℃ + -1.0 ℃), and the time from the start of the swimming of the mice to death was recorded as the mouse weight-bearing swimming time.
1.5.2 serum urea nitrogen assay: after 10 mice in each dose group of the subgroup 2 are subjected to final administration of the test substance for 30min, swimming for 90min in water at 30 ℃ without load, taking blood after resting for 60min, separating serum, and measuring serum urea nitrogen by using a Hitachi 7020 full-automatic biochemical analyzer, wherein a MAKER biochemical kit is adopted as a reagent.
1.5.3 liver glycogen determination: after 10 mice in each dose group of subgroup 3 are given the test substance for 30min last time, animals are sacrificed, livers are rinsed by normal saline and sucked by filter paper, 0.1-0.2g of livers are accurately weighed, and detection is carried out according to the instruction of the kit.
1.5.4 lactic acid assay: after 10 mice in each dose group of the 4 th subgroup last irrigate the stomach for 30min, 1-2mm of the mice are cut, the cotton balls are used for removing first drop of blood, trace tail blood is collected, and the lactic acid content of the blood in the resting state of the mice is measured by a lactic acid tester. Then the mice were swim for 10min in water at 30 ℃ without load, and the lactic acid value of the mice immediately after exercise and after rest was detected by a lactic acid tester at 0min and 20min after swimming, respectively.
1.6 statistics of test data: comparing the differences among animal weights, urea nitrogen, hepatic glycogen, blood lactic acid (before swimming, after swimming for 0min, after swimming for 20 min) and negative control groups by adopting an analysis of variance method, wherein p <0.05 is statistically significant; after reciprocal transformation and COS transformation are adopted respectively due to uneven variance of the load swimming time and the area under the blood lactic acid curve, variance analysis is used for comparing the differences between each experimental group and the negative control group, and p <0.05 is statistically significant.
1.7, judging the result: weight bearing swimming time: if the load swimming time of the tested sample group is obviously longer than that of the control group and the difference is significant, the test result can be judged to be positive; urea nitrogen: if the serum urea of the tested sample group is lower than that of the negative control group and the difference is significant, the test result can be judged to be positive; liver glycogen: if the liver glycogen content of the tested sample group is obviously higher than that of the negative control group and the difference is obvious, the test result can be judged to be positive; blood lactic acid: the area under the blood lactic acid curve at three time points is taken as a judgment standard. The area of any test group is smaller than that of the negative control group, and the difference is significant, so that the positive test result can be judged. The test result of the load swimming is positive, and any two of three biochemical indexes of blood lactic acid, serum urea and hepatic glycogen are positive, so that the tested sample can be judged to have the function of relieving physical fatigue.
2 results:
2.1 animal weight gain
From Table 6, it can be seen that the second and fourth subgroups B high dose groups had significant effects (p < 0.05) during the oyster peptide energy drink feeding, and that there was no significant difference between the remaining components compared to the negative control, thereby considering that the oyster peptide energy drink and the red cow vitamin drink's stomach lavage were not the major factors affecting the significant weight gain of mice, and that there was no dose-response relationship therebetween. In conclusion, oyster peptide energy drink and red cow vitamin drink have no influence on the weight of mice in the 30-day gastric lavage experiment.
TABLE 6 Effect of oyster peptide energy drink on weight gain in mice
Note that: * Indicating that the difference was significant compared to the negative control group (p < 0.05);
* *. shows that the difference was significant compared to the negative control group (p < 0.01).
A. Oyster peptide energy beverage
B. Table 7 effect of oyster peptide energy drink on mouse load swimming time
Note that: * Shows that the difference compared with the negative control group has significance (p < 0.01)
Animal experiment research shows that: the weight-bearing swimming time of the oyster peptide energy drink low-dose group and the red cattle vitamin flavor drink in the dose group is obviously higher than that of the negative control group (p is less than 0.01) after the continuous 30-day gastric administration of the mice, the weight-bearing swimming time of the oyster peptide energy drink low-dose group is 8.65+/-3.75 min and 7.66+/-3.16 min of the oyster peptide energy drink high-Yu Gongniu vitamin energy drink, and the weight-bearing swimming time of the mice is obviously prolonged.
TABLE 8 Effect of oyster peptide energy drink on mouse serum Urea Nitrogen and liver glycogen
Note that: ** shows that the difference compared with the negative control group has significance (p<0.01)。
After the oyster peptide energy drink and the red cow vitamin drink are infused with stomach for 30 days, compared with a negative control group, the high-dose group mice swim for 90 minutes without load, rest for 60 minutes, and serum urea nitrogen level is obviously reduced (p < 0.01); oyster peptide energy drink high dose group serum urea nitrogen 7.3+ -1.3 mmol/L red ox vitamin flavor drink 7.9+ -2.2 mmol/L (see Table 8).
Influence of oyster peptide energy drink on liver glycogen: the oyster peptide energy drink is given for 30 days, and compared with the negative control group, the liver glycogen level of the mice in each dose group has no obvious change; the liver glycogen levels of mice in the medium and high dose groups were significantly reduced compared to the negative control group by administering vitamin-flavored red cows for 30 days, and the differences were significant (p < 0.01) (see table 8).
TABLE 9 Effect of oyster peptide energy drink on lactic acid in blood of mice (mmol/L)
Note that: * shows that the difference compared with the negative control group has significance (p<0.05)；
** Shows that the difference compared with the negative control group has significance (p<0.01)。
a After logarithmic conversion, the data are normally distributed, each group of variance is uniform, and single-factor variance analysis is adopted;
b after logarithmic conversion, the data is randomly distributed in a non-normal way, and rank sum analysis is adopted.
Animal experiment research shows that: after 14.0, 46.7 and 140.0 ml/kg.BW oyster peptide energy drink is continuously and gastrically administrated to the mice for 30 days, the load swimming time of the mice is obviously prolonged; serum urea nitrogen levels are significantly reduced; the area under the blood lactic acid curve is obviously reduced at three time points of 0min before swimming and 20min after swimming; but has no obvious effect on liver glycogen levels in resting states in mice. According to the judging standard of the health department of health food inspection and evaluation technical Specification (2003 edition) for relieving physical fatigue, the marine active peptide energy drink can be considered to have the function of relieving physical fatigue.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and any person skilled in the art may make modifications or alterations to the above technical content to the equivalent embodiments. However, any simple modification, equivalent variation and variation of the above embodiments according to the technical substance of the present invention still fall within the protection scope of the technical solution of the present invention.
Claims (6)
1. A preparation method of oyster peptide is characterized in that the method is to carry out enzymolysis on edible parts of oyster, and the selected enzymes are animal compound protease and neutral protease.
2. The preparation method according to claim 1, characterized in that it comprises the following steps: freezing and homogenizing fresh oyster edible parts, adding 0.05-0.3 wt% of animal compound protease and 0.01-0.2 wt% of neutral protease, and performing enzymolysis for 1-7 h to obtain oyster peptide crude extract; and (3) decoloring the peptide crude extract, filtering, desalting the filtered filtrate, and spray-drying the desalted solution to obtain oyster peptide.
3. The method of claim 2, wherein the desalting is by membrane separation techniques.
4. Use of oyster peptide prepared according to claim 1 in a beverage product.
5. The use of oyster peptide prepared in claim 1 in energy beverage with anti-fatigue function.
6. An energy drink with an anti-fatigue function is characterized by comprising the following raw materials in percentage by mass: 75-90 parts of drinking water, 5-8 parts of oyster peptide prepared by the method of claim 1, 1-3 parts of natural taurine, 2-5 parts of inositol, 3-8 parts of nicotinamide, 0.02-0.05 part of vitamin B6 and 12 0.03-0.06 part and 1-3 parts of dietary fiber.
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