CN113637719A - A kind of chicken albumin oligopeptide and preparation method thereof - Google Patents
A kind of chicken albumin oligopeptide and preparation method thereof Download PDFInfo
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- CN113637719A CN113637719A CN202110916078.8A CN202110916078A CN113637719A CN 113637719 A CN113637719 A CN 113637719A CN 202110916078 A CN202110916078 A CN 202110916078A CN 113637719 A CN113637719 A CN 113637719A
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- oligopeptide
- chicken blood
- blood albumin
- albumin
- chicken
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- Peptides Or Proteins (AREA)
Abstract
The invention discloses a chicken blood albumin oligopeptide and a preparation method thereof, belonging to the technical field of food processing. According to the preparation method of the chicken blood albumin oligopeptide, the chicken blood albumin is subjected to enzymolysis in a specific enzymolysis process, so that the chicken blood albumin is fully degraded into small molecular polypeptides with activity, and then the small molecular polypeptides are further subjected to ultrafiltration interception and screening, so that the obtained oligopeptide product has high active substance content and has remarkable effects of resisting oxidation, enhancing organism immunity, resisting fatigue and the like; the preparation method has the advantages of simple production process, safety, no toxicity and high economic benefit, and can realize industrial large-scale production. The invention also provides the chicken blood albumin oligopeptide obtained by the preparation method, and the product has various biological activities and stronger activities of resisting oxidation, enhancing immunity, resisting fatigue and the like; the product has high purity, safety, no toxicity, and high economic use value. The invention also provides the application of the product in the preparation of medicines, foods and health products.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to chicken blood albumin oligopeptide and a preparation method thereof.
Background
China has huge broiler breeding and slaughtering processing industries, in recent years, the demand of domestic markets is greatly increased, the number of stocked broilers is about 10 billion every year, the stock production is as high as hundreds of billions, and China becomes the second largest chicken producing country in the world after the United states. The chicken blood is a byproduct in the process of slaughtering and processing broilers, the utilization rate of the chicken blood is not high at present in China, and part of the chicken blood is prepared into chicken blood powder which is used as animal feed, but the added value is low; the vast majority of the rest chicken blood is directly discharged into the environment as waste, which not only wastes resources but also causes serious environmental pollution. Blood contains various nutrients and bioactive substances, especially abundant proteins, and the content of proteins in blood plasma is about 8% of the total mass, while the content of proteins in blood cells is up to 35%. Therefore, the blood is an abundant protein resource library, and if people can continuously develop a new chicken blood protein utilization method, the economic value of the chicken blood can be fully exerted, the environmental pollution can be reduced, and the application prospect is wide. At present, more and more technologies are available for separating plasma protein powder and hemoglobin powder from chicken blood, but the chicken blood protein separation technology and application still deserve further development.
In recent years, bioactive peptides become the focus of research at home and abroad, and a great deal of research shows that oligopeptides generated by degrading animal and plant proteins, such as soybean peptide, corn peptide, hemoglobin peptide and the like, have various biological activities, such as blood pressure reduction, blood fat reduction, liver protection, immunity enhancement, intestinal flora regulation, oxidation resistance, bacteria resistance, virus resistance and the like, and are popular among consumers. Meanwhile, the oligopeptide can be directly absorbed and enter blood without being digested by gastrointestinal tracts, which has great significance for the elderly with the degradation of digestive functions, patients with gastrointestinal tract dysfunction and weak sub-health people. In view of the wide application prospect of small-molecule polypeptides, more and more oligopeptide production and preparation technologies and applications have been developed and disclosed. At present, the technology and process for preparing bioactive peptides are approaching to perfection, and the process mainly degrades animal and plant proteins by enzymolysis or fermentation to obtain oligopeptides. The proteolysis is the most common method for preparing polypeptide, and comprises a single enzyme method and a compound enzyme method, the reaction condition is mild and controllable, and the stability of the result can be ensured.
The plasma accounts for 65% of the whole blood in mass content, contains 8% of protein, and the albumin is the main protein component in the plasma and accounts for about 50% of the total protein in mass content. Mature chicken plasma albumin is a polypeptide chain consisting of 565 amino acid residues, is rich in various essential amino acids, particularly lysine, has the mass content of nearly 8 percent, and can be used as an amino acid supplement. Albumin has vital significance to the body, and has various physiological functions of maintaining the osmotic pressure of blood plasma, transporting important physiological substances in blood, providing nutrition for the body and the like. Clinically, human serum albumin can be used for treating shock and cerebral edema caused by blood loss and trauma and diseases such as intracranial pressure rise caused by injury. Research also shows that the serum albumin has better antioxidant activity. Therefore, the albumin in the chicken blood is utilized to prepare the bioactive peptide, so that the bioactive peptide has a better application prospect, not only can fully utilize abundant resources of the chicken blood to reduce waste, but also is beneficial to developing healthy food beneficial to human bodies, meets the requirements of consumers, and further improves the deep processing level of poultry breeding industry in China, the utilization of byproducts and the economic benefit of the industry.
In the prior art, CN110786496A discloses a preparation method and application of a meat flavor spice, and plasma protein powder extracted from poultry blood is taken as a raw material to prepare plasma protein peptide by enzymolysis; CN106119216A discloses a co-production process for extracting SOD and hemoglobin oligopeptide powder from livestock and poultry blood corpuscle liquid, and the technology discloses a scheme for preparing chicken hemoglobin oligopeptide by taking chicken blood as a raw material; CN102191304A discloses a preparation method of pig plasma antioxidant peptide, which prepares pig serum albumin reducing peptide through an enzymolysis process. However, there is no report on the utilization of chicken serum albumin to prepare oligopeptide products, and the related technologies are still under development and improvement.
Disclosure of Invention
Based on the defects of the prior art, the invention aims to provide the preparation method of the chicken blood albumin oligopeptide, the method fully utilizes active substances in the chicken blood, the production process is simple, the prepared product has high purity and activity, safety, no toxicity and high economic value, and has excellent biological activity in the aspects of oxidation resistance, organism immunity enhancement, fatigue resistance and the like.
In order to achieve the purpose, the invention adopts the technical scheme that:
a preparation method of chicken blood albumin oligopeptide comprises the following steps:
(1) dissolving the crude product of the chicken blood albumin in water to obtain a crude product mixed solution; the mass percentage of the chicken blood albumin in the mixed solution is 5-20%;
(2) adding protease into the crude product mixed solution, adjusting the pH value to 7.5-10.5, carrying out enzymolysis at 40-60 ℃ for 10-20 h, and then carrying out enzyme deactivation treatment; the addition amount of the protease is 0.3-1.5% (w/v) of the crude product mixed solution;
(3) centrifuging the mixed solution after enzyme deactivation, taking supernatant liquid for ultrafiltration and interception treatment, and drying to obtain the chicken serum albumin oligopeptide.
The chicken blood albumin is rich in various essential amino acids for human body, especially lysine, and has a mass content of approximately 8%, and can be used as amino acid supplement. Through repeated experimental research of the inventor, the chicken blood albumin oligopeptide product obtained by catalysis under the screened enzymolysis condition has high active peptide content, each essential amino acid is fully reserved and has high activity, and the chicken blood albumin oligopeptide product has obvious effects of resisting oxidation and fatigue and improving immunity.
The ultrafiltration interception technology used in the invention has a good separation effect on the small molecular active peptide obtained after enzymolysis, belongs to a physical separation method, does not introduce chemical reagents, is safe and nontoxic in the whole process, and guarantees the biological activity of the oligopeptide product to the maximum extent.
According to the preparation method of the chicken blood albumin oligopeptide, the chicken blood albumin is subjected to enzymolysis in a specific enzymolysis process, so that the chicken blood albumin is fully degraded into small molecular polypeptides with activity, and then the small molecular polypeptides are further subjected to ultrafiltration interception and screening, so that the obtained oligopeptide product has high active substance content and has remarkable effects of resisting oxidation, enhancing organism immunity, resisting fatigue and the like; the preparation method has the advantages of simple production process, safety, no toxicity and high economic benefit, and can realize industrial large-scale production.
Preferably, the mass percentage of the chicken blood albumin in the mixed solution in the step (1) is 12-16%.
The initial concentration can ensure that the concentration of the substrate in the crude raw material is moderate, thereby being beneficial to subsequent full enzymolysis and simultaneously avoiding the influence on the enzymolysis quality caused by excessive impurity of the chicken blood albumin contained in the mixed solution.
Preferably, the protease comprises at least one of alkaline protease, trypsin, papain, and flavourzyme.
The protease can be used for the biological enzymolysis process of the chicken blood albumin, the enzymolysis effect is good, and the activity of the small molecular peptide in the product is high.
Preferably, the pH value in the step (2) is 8.5-9.5.
Preferably, the temperature in the step (2) is 50-55 ℃.
Preferably, the addition amount of the protease in the step (2) is 0.5-1.0% of the mass percentage of the crude product mixed solution.
The conditions of pH value, temperature and enzyme addition amount are favorable for ensuring the sufficiency of the enzymolysis process: if the pH value and the temperature are too low or too high in the enzymolysis process, the activity of the protease can be changed; if the activity of the protease is too low, the enzymolysis process is not complete, and the chicken blood albumin may be denatured, which affects the purity and activity of the enzymolysis product. The addition amount of the protease in the enzymolysis process is also important for the enzymolysis quality, and if the addition amount of the protease is small, the enzymolysis efficiency in the same time is too low, so that the industrial production is not facilitated, and the raw material waste is caused; when the addition amount of the enzyme exceeds a certain limit, the exposure amount of some hydrophobic amino acid residues is possibly increased, and further certain bitter taste is generated in the oligopeptide product, so that the application of the oligopeptide product in the food industry is limited, and the process cost is increased.
Preferably, the enzymolysis time in the step (2) is 12-15 h.
After repeated experiments, the inventor finds that the oligopeptide product prepared under the preferable enzymolysis time has higher content of small molecular peptides, high production efficiency and less bad flavor, and can be used for preparing various foods.
Preferably, the ultrafiltration of step (3) retains polypeptide molecules having a molecular weight of less than 3000 Da.
The small molecular peptide retained under strict screening has high content, less impurities, good water solubility and high purity of essential amino acid contained in the small molecular peptide.
Preferably, the drying of step (3) is spray drying.
More preferably, the inlet temperature of the spray drying is 160-180 ℃, and the outlet temperature is 60-80 ℃.
The spray drying under the condition can fully remove the water of the oligopeptide product prepared from the chicken blood albumin, so that the purity of the product is higher.
The invention also aims to provide the chicken blood albumin oligopeptide prepared by the preparation method of the chicken blood albumin oligopeptide.
The chicken blood albumin oligopeptide has various biological activities, has strong antioxidant activity, and simultaneously has the activities of enhancing immunity, resisting fatigue and the like; the product has high purity, safety, no toxicity, and high economic use value.
The invention also aims to provide the application of the chicken blood albumin oligopeptide in the preparation of medicines, foods and health-care products.
The oligopeptide product has high stability, safety, no toxicity and rich nutrition, and can be fully applied to the preparation of various edible products.
The preparation method of the chicken blood albumin oligopeptide has the beneficial effects that the chicken blood albumin is subjected to enzymolysis in a specific enzymolysis process to be fully degraded into small molecular polypeptides with activity, and then the small molecular polypeptides are screened by further ultrafiltration interception, so that the obtained oligopeptide product has high active substance content and has remarkable effects of resisting oxidation, enhancing the immunity of the organism, resisting fatigue and the like; the preparation method has the advantages of simple production process, safety, no toxicity and high economic benefit, and can realize industrial large-scale production. The invention also provides the chicken blood albumin oligopeptide obtained by the preparation method, and the product has various biological activities and stronger activities of resisting oxidation, enhancing immunity, resisting fatigue and the like; the product has high purity, safety, no toxicity, and high economic use value. The invention also provides application of the chicken serum albumin oligopeptide in preparation of medicines, foods and health-care products.
Drawings
FIG. 1 is a schematic diagram showing the effect of the anti-oxidant activity of the chicken blood albumin oligopeptide in effect example 1 of the present invention;
FIG. 2 is a graph showing ORAC values of chicken blood albumin oligopeptide at different concentrations in example 1 of the present invention.
Detailed Description
Unless otherwise specified, the raw materials used in the examples of the present invention and comparative examples were commercially available, and the production equipment used was a commercially available common model.
For better illustrating the objects, technical solutions and advantages of the present invention, the present invention will be further described with reference to specific examples, which are intended to be understood in detail, but not intended to limit the present invention. The crude product of the chicken blood albumin is prepared according to the prior art (the preparation and property research of the chicken blood albumin, 1998 (01): 41-45, proceedings of Shantou university (Nature science edition)), and the properties of the raw material are matched with the records of the prior art.
Example 1
One embodiment of the preparation method of the chicken blood albumin oligopeptide is disclosed in the invention.
The preparation method of the chicken blood albumin oligopeptide comprises the following steps:
(1) dissolving 100g of crude chicken blood albumin in water to obtain a crude mixed solution; the mass percentage of the chicken blood albumin in the mixed solution is 10 percent;
(2) adding alkaline protease into the crude product mixed solution, adjusting pH to 8.5 with NaOH, performing enzymolysis at 55 deg.C for 15 hr, and inactivating enzyme at 95 deg.C for 10 min; the addition amount of the protease is 1% (w/v) of the crude product mixed solution;
(3) cooling to room temperature, centrifuging the mixed solution after enzyme deactivation for 15min at the speed of 4000rpm, taking supernatant as enzymolysis liquid, performing ultrafiltration interception treatment, intercepting and separating oligomeric polypeptide molecules with the molecular weight of less than 3000Da in the enzymolysis liquid, and performing spray drying on ultrafiltrate to obtain 76.9g of the chicken blood albumin oligopeptide; the inlet temperature of the spray drying is 170 ℃ and the outlet temperature is 80 ℃.
The product has good water solubility, no other bad flavor except slight bitter taste; the components of the obtained chicken blood albumin oligopeptide are detected, the test method is a Kjeldahl method (GB/T5009.5-2003), and the detection shows that the product has the active peptide content of 85.6 percent by mass, the water content of only 5.3 percent by mass and the ash content of less than 1.3 percent by mass, which indicates that the prepared product has high purity.
The molecular weight distribution of the product is shown in table 1, and it can be known from the table that macromolecular substances are basically filtered after ultrafiltration interception treatment, and active ingredients in the product are mainly micromolecular oligopeptide with the molecular weight of less than 3000Da or even less than 2000 Da.
TABLE 1
The amino acid content of the prepared chicken blood albumin oligopeptide was analyzed by using an automatic amino acid analyzer, the technical parameters used in the analysis were the same as those in the preparation and property research of chicken blood albumin (1998 (01): 41-45; published by Shantou university (Nature science edition)), and the test results are shown in Table 2.
TABLE 2
| Amino acids | Content (g/100g) |
| Lysine | 7.03 |
| Phenylalanine | 4.96 |
| Methionine | 3.04 |
| Threonine | 2.68 |
| Isoleucine | 4.85 |
| Leucine | 6.52 |
| Valine | 4.69 |
| Alanine | 3.42 |
| Arginine | 4.66 |
| Asparagine | 2.23 |
| Aspartic acid | 5.41 |
| Cysteine | 4.85 |
| Glutamine | 5.12 |
| Glutamic acid | 7.09 |
| Glycine | 1.8 |
| Histidine | 1.84 |
| Tyrosine | 3.68 |
| Proline | 3.25 |
| Serine | 4.31 |
As can be seen from the table, the essential amino acids with high activity in human body in the chicken blood albumin oligopeptide prepared by the embodiment are higher in content, wherein the content of lysine in the essential amino acids is as high as 7.03% by mass.
Example 2
One embodiment of the preparation method of the chicken blood albumin oligopeptide is disclosed in the invention.
The preparation method of the chicken blood albumin oligopeptide comprises the following steps:
(1) dissolving 100g of crude chicken blood albumin in water to obtain a crude mixed solution; the mass percentage of the chicken blood albumin in the mixed solution is 8 percent;
(2) adding trypsin into the crude mixed solution, adjusting pH to 7.4 with NaOH, performing enzymolysis at 50 deg.C for 18 hr, and inactivating enzyme at 95 deg.C for 10 min; the addition amount of the protease is 1.5 percent (w/v) of the crude product mixed solution;
(3) cooling to room temperature, centrifuging the mixed solution after enzyme deactivation for 15min at the speed of 4000rpm, taking supernatant as enzymolysis liquid, performing ultrafiltration interception treatment, intercepting and separating oligomeric polypeptide molecules with the molecular weight of less than 3000Da in the enzymolysis liquid, and performing spray drying on ultrafiltrate to obtain 74.3g of the chicken blood albumin oligopeptide; the inlet temperature of the spray drying is 170 ℃ and the outlet temperature is 80 ℃.
The product has good water solubility, no other bad flavor except slight bitter taste; the components of the obtained chicken blood albumin oligopeptide are detected, the testing method is the same as that in example 1, and the detection proves that the product has the active peptide content of 70.6 percent by mass, the water content of only 6.2 percent by mass and the ash content of less than 1.8 percent by mass, which indicates that the prepared product has high purity.
Effect example 1
To verify the antioxidant activity of the chicken serum albumin oligopeptide prepared by the invention, the product obtained in example 1 is subjected to an Oxidative Radical Absorption Capacity (ORAC) detection experiment. The detection steps are as follows: four experimental variable groups of AAPH-group (blank), AAPH + group (positive control), high concentration test sample group (4mg/mL) and low concentration test sample group (2mg/mL) were set, and Trolox group (standard) was set at the same time. PBS, the test sample prepared in example 1, sodium fluorescein and AAPH were added to a 96-well plate and rapidly placed in a fluorescence microplate reader to detect the change in fluorescence signal (at 485nm excitation and 535nm emission) within 2h at 37 ℃. Since fluorescein sodium is easily quenched by free radical attack generated by AAPH, the antioxidant capacity of the tested sample can be reflected by detecting the change of fluorescence intensity. The test results are shown in fig. 1 and 2, and it is evident that the chicken blood albumin oligopeptide product prepared in example 1 has significant antioxidant activity, and the oxidative radical absorption capacity is correspondingly improved with the increase of the concentration.
Effect example 2
To verify that the chicken serum albumin oligopeptide prepared by the invention has an effect of improving the immune function, the following test experiments are carried out on the product obtained in example 1:
(1) thymus and spleen index test
The thymus is a central immune organ of an organism and is an important place for the differentiation and maturation of T cells; the spleen is the important peripheral immune organ of the body, and the site for the colonization and proliferation of T lymphocytes and B lymphocytes. The thymus index and the spleen index can reflect the immune function of an organism, and the influence of a test object on the immunity of the organism can be detected by observing the thymus index and the spleen index. On the other hand, immunosuppressive agents such as cyclophosphamide inhibit lymphocyte differentiation, decrease the number of lymphocytes in immune organs such as thymus and spleen, and further lead to a decrease in the weight of immune organs. Therefore, the experiment establishes a model control group through cyclophosphamide, and comprises the following specific steps: 35 Kunming mice were randomly divided into a normal group, a Cyclophosphamide (CTX) model group, a chicken blood albumin oligopeptide low dose group (25mg/kg) prepared in example 1, a chicken blood albumin oligopeptide medium dose group (50mg/kg) prepared in example 1, and a chicken blood albumin oligopeptide high dose group (100mg/kg) prepared in example 1, and 7 mice were each group. One week after the animals of each group were acclimatized, the corresponding groups were continuously gavaged with different doses of albumin oligopeptide for 21 days. From the 19 th day of administration, except the normal group, each group was intraperitoneally injected with 80mg/kg cyclophosphamide daily to establish an immunosuppressive mouse model. After the last administration, the mice fasting for 24h, weighing the weight, taking blood from the orbit, and taking the thymus and spleen for experimental detection. The mouse thymus and spleen were weighed, and the corresponding organ index (%) -organ weight (g)/body weight (g) was calculated. The results of the mouse thymus index and spleen index tests are shown in table 3.
TABLE 3
Compared with the blank control group, the composition of the composition,*P<0.05; compared with the cyclophosphamide model group,#P<0.05。
as is apparent from table 3, the thymus index and spleen index of the cyclophosphamide model group without the oligopeptide product injection were significantly decreased compared to the blank control group, whereas in contrast, the two indices of the variable group with the oligopeptide product described in example 1 were increased, and the higher the dose, the larger the increase.
(2) Test for promoting increase in leukocyte count
When pathogenic bacteria invade the human body, white blood cells can penetrate through the capillary wall through deformation, migrate from the inside of the blood vessel to the tissues outside the blood vessel and concentrate to the invasion part of the pathogenic bacteria to surround and phagocytose the pathogenic bacteria, and the quantity of the white blood cells reflects the immunity of the organism. Therefore, the number of leukocytes in peripheral blood is an important index for detecting the immune function of the body. The orbit of each group of mice in the detection test (1) was bled, the number of leukocytes in the blood was detected using a fully automatic blood cell analyzer, and the average value of the number of each group was finally counted. The results of the measurement of the peripheral blood leukocyte count of each group of mice are shown in Table 4.
TABLE 4
| Group of | Number of leukocytes (. times.10)9/L) |
| Blank control group | 7.17±0.81 |
| Cyclophosphamide model group | 4.25±0.76* |
| Oligopeptide Low dose group | 4.81±0.36 |
| Oligopeptide medium dose group | 5.24±0.36# |
| Oligopeptide high dose group | 6.13±0.29# |
P <0.05 compared to placebo; compared to the cyclophosphamide model group, # P < 0.05.
As can be seen from Table 4, compared with the blank control group, the addition of cyclophosphamide in the remaining four groups significantly reduced the number of white blood cells of the mice, while the numbers of white blood cells in the blood of the mice in the medium-dose and high-dose groups using the medium-high dose oligopeptide product of the present invention were significantly increased, which indicates that the chicken blood albumin oligopeptide product of the present invention can effectively increase the number of white blood cells in the blood.
(3) Macrophage phagocytosis rate assay
Macrophages are multifunctional immune cells, and their phagocytic function is an important component of nonspecific immunity in the body. In addition, macrophages also have cytokine secretion and antigen presentation functions, thereby indirectly participating in the immune response of the body. Therefore, the phagocytic function of macrophages directly reflects the immune status of the body. The experiment detects the phagocytic capacity of the abdominal cavity macrophages according to the method that the abdominal cavity macrophages of the mice phagocytize the chicken red blood cells. The method comprises the following specific steps: 35-Kunming mice were randomly divided into a normal group, a Cyclophosphamide (CTX) model group, a low dose group (25mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, a medium dose group (50mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, and a high dose group (100mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, and 7 mice were each group. One week after the animals of each group were acclimatized, the corresponding groups were continuously gavaged with different doses of albumin oligopeptide for 21 days. From the 19 th day of administration, except the normal group, each group was intraperitoneally injected with 80mg/kg cyclophosphamide daily to establish an immunosuppressive mouse model. After the last administration, 0.5mL of 3% chicken red blood cell suspension was injected into the abdominal cavity of each mouse, and the suspension was dispersed by lightly pressing the abdomen of the mouse. After 60min, collecting the abdominal cavity liquid of the mice, and observing and recording the phagocytosis of chicken red blood cells by the macrophagic fines in the abdominal cavity through a microscope. The phagocytosis ratio (%) × 100% (number of macrophages engulfed chicken red blood cells/total number of macrophages), and the phagocytosis index (%) — total number of chicken red blood cells engulfed by macrophages/total number of macrophages engulfed chicken red blood cells.
The test results are shown in table 5.
TABLE 5
| Group of | Phagocytosis ratio (%) | Phagocytic index |
| Blank control group | 33.3±2.57 | 0.43±0.04 |
| Cyclophosphamide model group | 21.9±2.62* | 0.23±0.02* |
| Oligopeptide Low dose group | 23.4±2.25 | 0.25±0.02 |
| Oligopeptide medium dose group | 25.3±2.26# | 0.28±0.03# |
| Oligopeptide high dose group | 28.6±1.80# | 0.34±0.02# |
Compared with the blank control group, the composition of the composition,*P<0.05; compared with the cyclophosphamide model group,#P<0.05。
the results in table 5 show that, compared with the blank control group, the cyclophosphamide added in the other four groups of cyclophosphamide groups significantly reduces the phagocytic rate and phagocytic index of the mouse abdominal cavity macrophages, while the moderate-dose and high-dose groups using the oligopeptide product of the present invention significantly improve the phagocytic rate and phagocytic index of the mouse abdominal cavity macrophages, which indicates that the chicken blood albumin oligopeptide product of the present invention can effectively improve the phagocytic rate and phagocytic index of the macrophages in blood.
Effect example 3
To verify the anti-fatigue effect of the chicken serum albumin oligopeptide prepared by the invention, the following test experiments are carried out on the product obtained in example 1:
28 mice, one from Kunming, were randomly divided into a normal group, a low dose group (25mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, a medium dose group (50mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, and a high dose group (100mg/kg) of the chicken blood albumin oligopeptide prepared in example 1, each group consisting of 7 mice. One week after the animals of each group were acclimatized, the corresponding groups were continuously gavaged with different doses of albumin oligopeptide for 21 days. 30min after the last administration, 5% weight of lead skin load was applied to the tail root of each mouse, and the mice were placed in a 30 cm swimming chamber for exhaustive swimming experiments with water temperature controlled at 25 + -1 deg.C. The time from the beginning of swimming to the appearance of the exhaustion state is recorded as the mouse exhaustion swimming time. And sampling the blood and the liver of the mouse for subsequent experimental determination.
In the experimental test of the effect example, the load swimming test is a forced whole-body energy loss activity, and the swimming time of the load swimming test can reflect the anti-fatigue capability of a mouse; hepatic glycogen is an important energy storage mode of an organism, can be decomposed into glucose to maintain the stability of blood sugar, and during exhaustive exercise, hepatic glycogen is decomposed and metabolized into energy to be used by the organism, so that the content of hepatic glycogen is reduced; urea nitrogen is the end product of catabolism of amino acids via the ornithine cycle. When energy generated by sugar metabolism and fat metabolism cannot meet the needs of an organism due to long-time strenuous exercise, protein and amino acid participate in metabolism to provide energy; the organism is in anaerobic respiration state during exhaustion exercise, and rapid energy supply is carried out through glycolysis. Glucose is metabolized into lactic acid, which is accumulated in muscle tissue, resulting in the decrease of muscle motor ability and fatigue of the body. Therefore, whether the product has the anti-fatigue gain effect or not can be accurately verified by detecting the indexes.
The results of the tests are shown in Table 6.
TABLE 6
Compared with the blank control group, the composition of the composition,*P<0.05。
from the results in Table 7, it can be seen that the low, medium and high dose oligopeptide products significantly prolonged the swimming time of mice, increased the liver glycogen levels of mice, and decreased the urea nitrogen and blood lactic acid levels of mice, compared to the blank control group. Therefore, according to the comprehensive evaluation of the indexes, the chicken blood albumin oligopeptide product has better anti-fatigue effect.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A preparation method of chicken blood albumin oligopeptide is characterized by comprising the following steps:
(1) dissolving the crude product of the chicken blood albumin in water to obtain a crude product mixed solution; the mass percentage of the chicken blood albumin in the mixed solution is 5-20%;
(2) adding protease into the crude product mixed solution, adjusting the pH value to 7.5-10.5, carrying out enzymolysis at 40-60 ℃ for 10-20 h, and then carrying out enzyme deactivation treatment; the addition amount of the protease is 0.3-1.5% (w/v) of the crude product mixed solution;
(3) centrifuging the mixed solution after enzyme deactivation, taking supernatant liquid for ultrafiltration and interception treatment, and drying to obtain the chicken serum albumin oligopeptide.
2. The method for preparing the chicken blood albumin oligopeptide according to claim 1, wherein the mass percentage of the chicken blood albumin in the mixed solution in the step (1) is 12-16%.
3. The method of claim 1, wherein the protease comprises at least one of alkaline protease, trypsin, papain, and flavourzyme.
4. The method for preparing chicken blood albumin oligopeptide according to claim 1, wherein the pH in the step (2) is 8.5-9.5.
5. The method for preparing the chicken blood albumin oligopeptide according to claim 1, wherein the temperature in the step (2) is 50-55 ℃.
6. The method for preparing chicken blood albumin oligopeptide according to claim 1, wherein the protease is added in the step (2) in an amount of 0.5-1.0% by mass of the crude mixed solution.
7. The method for preparing chicken blood albumin oligopeptide according to claim 1, wherein the enzymolysis time in the step (2) is 12-15 h.
8. The process for the preparation of chicken blood albumin oligopeptide according to claim 1, wherein the ultrafiltration cut-off of step (3) is performed by spray drying, wherein the cut-off of the polypeptide molecules is less than 3000 Da.
9. The chicken blood albumin oligopeptide as set forth in any one of claims 1 to 8.
10. The use of the chicken blood albumin oligopeptide of claim 9 in the preparation of medicaments, foods and health products.
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