CN114989247B - Anti-hypoxia or hypoxia anti-fatigue peptide prepared from yak milk residues, and preparation method and application thereof - Google Patents

Anti-hypoxia or hypoxia anti-fatigue peptide prepared from yak milk residues, and preparation method and application thereof Download PDF

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CN114989247B
CN114989247B CN202210453879.XA CN202210453879A CN114989247B CN 114989247 B CN114989247 B CN 114989247B CN 202210453879 A CN202210453879 A CN 202210453879A CN 114989247 B CN114989247 B CN 114989247B
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CN114989247A (en
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孙术国
杨飞艳
何旭东
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Central South University of Forestry and Technology
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Abstract

The invention relates to an anti-hypoxia or hypoxia and anti-fatigue peptide of yak milk residues, and a preparation method and application thereof, wherein the anti-hypoxia or hypoxia and anti-fatigue peptide of the yak milk residues is respectively T1, T2 and T3, T1 has an amino acid sequence shown as SEQ ID NO.1, the molecular weight is 1000.50Da, T2 has an amino acid sequence shown as SEQ ID NO.2, the molecular weight is 1822.00Da, T3 has an amino acid sequence shown as SEQ ID NO.3, and the molecular weight is 1212.65Da. The invention has very good anti-hypoxia or hypoxia and anti-fatigue effects, can improve the integral value of Tibetan milk and dairy products, and can improve the income of farmers and pastures.

Description

Anti-hypoxia or hypoxia anti-fatigue peptide prepared from yak milk residues, and preparation method and application thereof
Technical Field
The invention belongs to the field of biological micromolecular active peptides, relates to an anti-hypoxia or hypoxia and anti-fatigue peptide derived from yak milk residues, and in particular relates to an anti-hypoxia or hypoxia and anti-fatigue peptide of the yak milk residues, and a preparation method and application thereof.
Background
The energy required for the survival of the human body is mainly produced by oxidation reaction, and oxygen is essential in maintaining the vital activity of the body. However, the human body often causes hypoxia or anoxia of the human body due to diseases, mental stress, altitude hypoxia environment and other factors, thus further causing damages such as abnormal metabolism, functions and morphological structures of human tissues, and the like, and being life threatening in severe cases. In order to reduce the injury of human body in the state of hypoxia or anoxia, the effective measure is to inject or eat the anti-hypoxia or hypoxia anti-fatigue medicine into human body. At present, two main classes of anti-hypoxia or hypoxia anti-fatigue medicines are available on the market, wherein one class is western medicines comprising acetazolamide, methazolamide, dexamethasone, medroxyprogesterone, theophylline and sumatriptan; the other is the plant and its extract containing antioxidant active components, including semen Ginkgo and its extract, semen Strychni extract, radix Rhodiolae and its extract, turnip polysaccharide, saviae Miltiorrhizae radix and its preparation, extract of Yunnan Bulbus Lilii of Physcomitrella, herba Saussureae Involueratae and its extract, ginsenoside, quercetin, etc. The former chemical drugs have certain relief effect on symptoms caused by hypoxia, but have great side effects, and the drugs have high accumulation in organs and are easy to cause serious immune response, so the practical use is greatly limited; the natural products of the latter plants are mostly derived from rare traditional Chinese medicinal materials, and the extraction, separation and purification processes are complex, so that the price of the medicines is high. Therefore, the development of the anti-hypoxia or hypoxia anti-fatigue medicine with homology of medicine and food has small side effect, definite curative effect and high cost performance.
At present, with the rapid development of polypeptide histology, polypeptide substances are receiving more and more attention in disease treatment due to various biological characteristics. Currently, researchers develop anti-myocardial ischemia and hypoxia polypeptides (CN 201910790480.9 and CN 201910792484.0) in the aspect of researching and treating coronary atherosclerotic heart disease, however, the source information of the polypeptides is lacking, and the clinical data of human bodies is lacking, so that the anti-hypoxia effect of the anti-myocardial ischemia and hypoxia polypeptides is to be confirmed. Researchers also develop giant salamander active peptide compositions (CN 201911344308.7) with the functions of resisting aging, regulating blood pressure and resisting hypoxia, wherein the active peptide accounts for a small proportion in a formula, and although a mouse experiment proves that the giant salamander active peptide composition has the effect of resisting hypoxia, because the composition has complex components, whether the giant salamander active peptide has the effect of resisting hypoxia is difficult to judge. Yak is an animal special for low oxygen in suitable highland in China, in order to comprehensively utilize yak blood, researchers develop anti-hypoxia peptide from yak blood (Shora, research on anti-hypoxia effect of yak blood anti-hypoxia peptide and action mechanism thereof (D), yagan, university of Sichuan agriculture, 2019) and research on mechanism thereof, however, the structure of the researched anti-hypoxia peptide is not clear and lacks data in clinical aspects of human body. The milk residue is prepared by heating the defatted milk obtained by extracting the whole yak milk to prepare butter, adding abomasum enzyme or yoghourt, filtering and drying in the sun to obtain byproducts, and the milk residue is rich in protein, amino acid, calcium, iron, vitamins, lactose content and the like, however, due to technical limitations, the milk residue protein is not fully utilized, and resource waste is caused.
Disclosure of Invention
The invention provides an anti-hypoxia or hypoxia and anti-fatigue peptide of yak milk residues, a preparation method and application thereof, and develops the anti-hypoxia peptide with a definite structure and good health care effect.
In order to achieve the purpose, the invention adopts the following technical scheme:
the anti-hypoxia or hypoxia anti-fatigue peptide of the yak milk residues is respectively T1, T2 and T3, wherein T1 has an amino acid sequence shown as SEQ ID NO.1, the molecular weight is 1000.50Da, T2 has an amino acid sequence shown as SEQ ID NO.2, the molecular weight is 1822.00Da, T3 has an amino acid sequence shown as SEQ ID NO.3, and the molecular weight is 1212.65Da.
Preferably, the raw material of the yak milk residue anti-hypoxia or hypoxia anti-fatigue peptide is yak milk in the area with the altitude of 2800-5500 m.
A method for preparing anti-hypoxia or hypoxia anti-fatigue peptide from yak milk residues, which comprises the following steps of,
1) The yak milk residue sampling method comprises the following steps: adopting the yak milk residue powder or milk residue blocks freeze-dried by natural wind under the conditions of the altitude of the Tibetan plateau of 4000m, the winter temperature of minus 10 ℃ to minus 25 ℃ and the low oxygen concentration of 12 percent and the natural average wind speed of 2.5 m/s;
2) Pulverizing the yak milk residue powder or the milk residue blocks obtained in the step 1) to obtain yak milk residue fine powder;
3) Preparing the anti-hypoxia or hypoxia and anti-fatigue peptide from yak milk residues: adding distilled water and enzyme into the yak milk residue fine powder obtained in the step 2), carrying out ultrasonic crushing and enzymolysis, and carrying out freeze drying on the obtained zymolyte to obtain the yak milk residue anti-hypoxia or hypoxia anti-fatigue peptide freeze-dried powder.
Further preferably, the pulverizing conditions in the step 2) are as follows: 11500-12500rpm, and shearing for 4.5-5.5min.
Preferably, the drying temperature in the step 2) is as follows: air-drying at a low temperature of between-10 ℃ and-25 ℃.
Further preferably, the step 3) is carried out according to a feed liquid ratio of 1: distilled water is added into 2-4 g/ml, and the ratio of the enzyme addition amount to the yak milk residue fine powder mass is 1: (50-150), ultrasonic crushing, and carrying out water bath enzymolysis for 2-4 h at the temperature of 32-42 ℃.
Still further preferably, the enzyme of step 3) is pepsin+trypsin, the efficacy of pepsin being 1: 25000-35000, the enzyme activity of trypsin is more than 250USP u/mg.
Preferably, the step 3) is carried out for 2 to 4 hours each time, after the enzymolysis is finished each time, the supernatant is remained after centrifugation at 3000 to 5000rpm, the milk residue sediment is continuously added with the enzymolysis liquid for the next enzymolysis, and finally, all the supernatant is collected for analysis, and each time of enzymolysis is carried out for 1 to 3 times.
The application of the yak milk residue anti-hypoxia or hypoxia and anti-fatigue peptide or the yak milk residue anti-hypoxia or hypoxia and anti-fatigue peptide obtained by the preparation method of the yak milk residue anti-hypoxia or hypoxia and anti-fatigue peptide in anti-hypoxia drugs, foods or health products.
The invention has the beneficial effects that:
the anti-hypoxia or hypoxia and anti-fatigue peptide obtained by the method is derived from yak milk residues, and whether the yak milk residues are subjected to anti-hypoxia or hypoxia and anti-fatigue peptide freeze-dried powder or separated and purified to obtain high-purity T1, T2 and T3, has very good anti-hypoxia or hypoxia and anti-fatigue effects, so that the anti-hypoxia or hypoxia and anti-fatigue peptide prepared by the method has the advantages of small development side effect, definite curative effect and high cost performance, and meanwhile, as the yak milk residues are widely sourced in a Tibetan area and are by-products of butter processing, the method can change waste into valuables, improve the integral value of milk and dairy products in the Tibetan area and improve the income of farmers.
Drawings
FIG. 1 is a picture of the morphology of the anti-hypoxia or hypoxia and anti-fatigue peptide T1 of the yak milk residues prepared by the invention on neural cell protection cells for 6 hours of hypoxia;
FIG. 2 shows the effect of the addition of the anti-hypoxia or hypoxia anti-fatigue peptide T1 to the hypoxia 6-hour nerve cell activity of the yak milk residues prepared by the invention;
FIG. 3 is a graph showing the morphology of the anti-hypoxia or hypoxia and anti-fatigue peptide T1 of the yak milk residues prepared by the invention on nerve cell protection cells for 12 hours under hypoxia;
FIG. 4 shows the effect of the addition of the anti-hypoxia or hypoxia anti-fatigue peptide T1 prepared by the invention on the activity of nerve cells in hypoxia for 12 hours;
FIG. 5 shows the protective effect of the anti-hypoxia or hypoxia anti-fatigue peptide T1 of the yak milk residues on hypoxia 24-hour nerve cells;
FIG. 6 shows the effect of the addition of the anti-hypoxia or hypoxia anti-fatigue peptide T1 to the hypoxia 24-hour nerve cell activity of the yak milk residues prepared by the invention;
FIG. 7 is a picture of the morphology of the anti-hypoxia or hypoxia and anti-fatigue peptide T2 of the yak milk residues prepared by the invention on neural cell protection cells for 6 hours of hypoxia;
FIG. 8 shows the effect of the addition of anti-hypoxia or hypoxia anti-fatigue peptide T2 to the yak milk residues prepared by the invention on the activity of nerve cells in hypoxia for 6 hours;
FIG. 9 shows the protective effect of the anti-hypoxia or hypoxia anti-fatigue peptide T3 of the yak milk residues prepared by the invention on nerve cells in hypoxia for 6 hours;
FIG. 10 shows the effect of the addition of anti-hypoxia or hypoxia anti-fatigue peptide T3 to hypoxia 6 hr nerve cell activity of yak milk residues prepared in accordance with the present invention;
FIG. 11 shows the protective effect of the anti-hypoxia or hypoxia anti-fatigue peptide T3 of the yak milk residues prepared by the invention on nerve cells in hypoxia for 12 hours;
FIG. 12 shows the effect of the addition of anti-hypoxia or hypoxia anti-fatigue peptide T3 to hypoxia 12 hr nerve cell activity;
FIG. 13 shows the protective effect of the anti-hypoxia or hypoxia anti-fatigue peptide T3 of the yak milk residues prepared by the invention on nerve cells in hypoxia for 24 hours;
FIG. 14 shows the effect of the addition of anti-hypoxia or hypoxia anti-fatigue peptide T3 to hypoxia 24 hr nerve cell activity of yak milk residues prepared in accordance with the present invention.
The specific embodiment is as follows:
the invention will be further illustrated with reference to specific examples. The present invention will be described in further detail with reference to examples, but is not limited to these examples.
Example 1
A preparation method of a yak milk residue anti-hypoxia or hypoxia anti-fatigue peptide comprises the following steps:
1) The yak milk residue sampling method comprises the following steps: selecting yak milk in a region with an altitude of 2800-5500 m, preparing milk residue in the process of processing butter by a traditional process, further air-drying at a low temperature below 0 ℃ to obtain milk residue powder or milk residue blocks, preferably milk residue powder, wherein the storage time is within 1 month, the storage time is more than 1 month under the condition of 0-4 ℃, and the storage time is kept for standby under the condition of-18 ℃;
2) Adopting the yak milk residue powder or the milk residue blocks in the step 1), and shearing and crushing for 5min at a high speed of 12000rpm by a crusher to obtain yak milk residue fine powder;
3) Preparing the anti-hypoxia or hypoxia and anti-fatigue peptide from yak milk residues: and (3) mixing the yak milk residue fine powder obtained in the step (2) according to a feed liquid ratio of 1: distilled water was added at 3g/ml, and the ratio of enzyme addition to substrate was 1:100, ultrasonic crushing, carrying out enzymolysis for 3 hours in a 35 ℃ water bath, and freeze-drying the obtained zymolyte to obtain freeze-dried peptide powder of the anti-hypoxia or hypoxia and anti-fatigue peptide of the yak milk residue, and preserving at low temperature;
4) The yak milk residue anti-hypoxia or hypoxia and anti-fatigue peptide freeze-dried powder prepared in the step 3) is identified by adopting a polypeptide group, three anti-hypoxia or hypoxia and anti-fatigue peptides which are respectively T1, T2 and T3, the amino acid sequence YPFGPPN of T1, the molecular weight of the peptide is 1000.50Da, the amino acid sequence PVVVPPFLQPEVMGVSK of T2, the molecular weight of the peptide is 1822.00Da, the amino acid sequence LVYPFPGPIPN of T3, the molecular weight of the peptide is 1212.65Da, according to the molecular weight of T1, T2 and T3, the purity of the peptide is more than 95 percent of T1, T2 and T3 respectively by adopting a separation and purification conventional technical means, and further identification and confirmation are carried out by HPLC-MS-MS and nuclear magnetic resonance, and the anti-hypoxia or hypoxia and anti-fatigue peptide freeze-dried powder of T1, T2 and T3 are detected and analyzed, and applied to anti-hypoxia health foods.
Further, in the step 3), in the preparation method of the yak milk residue polypeptide, the added enzyme is pepsin+trypsin, and the efficiency of the pepsin is 1:30000 trypsin has an enzymatic activity of more than 250USP u/mg.
Further, in the step 3), in the preparation method of the yak milk residue polypeptide, the enzymolysis is carried out for 3 hours each time, and the enzymolysis is carried out for 2 times.
In order to make the objects and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the following cell experiments, animal experiments, and human clinical experiments. It should be understood that the specific experiments described herein are only for the purpose of illustrating the invention and are not to be construed as limiting the invention.
Unless otherwise specified, all reagents used in the following experiments were commercially available, and the methods of operation were all conventional.
Experimental results
Cell assay for protection of hypoxic nerve cells
1. Experimental cell
The experiments were performed using rat neuronal cell RSC96, available from the China academy of sciences typical culture Collection Committee cell Bank (National Collection of Authenticated Cell Cultures).
2. Cell hypoxia method
Experimental grouping: the method is divided into an anoxic control group, an anoxic experimental group and an normoxic control group.
Normoxic control group: cells were exposed to 95% CO 2 Normally cultured in a gaseous atmosphere for 24 hours (normoxic control group);
hypoxia control group: placing cells in 3%O 2 ，5％CO 2 And 92% N 2 Is cultured for 24 hours in an anoxic environment (anoxic control group);
hypoxia experimental group: cells were post-administration placed in 3%O 2 ，5％CO 2 And 92% N 2 Is cultured for 24 hours in an anoxic environment (anoxic experimental group: the dosage of the milk residue peptide is 25 mu g/mL,50 mu g/mL,100 mu g/mL). The results are shown in FIGS. 1-14.
As can be seen from fig. 1 to 14, in the anoxic model established by RSC96 of the blood-profound cells (nerve cells) of the rat, the toxicity MTS experimental result shows that the three polypeptides T1, T2 and T3 are not toxic to RSC96 cells, and the anoxic control group has a significantly reduced cell shape and a partially dead cell shape compared with the normoxic control group under the anoxic condition of RSC96 cell administration for 6 hours, 12 hours and 24 hours, and the anoxic control group has a significantly improved cell state, significantly reduced shrinkage cell and dead cell shape compared with the anoxic administration group of T1, T2 and T3, and is concentration dependent. In 6h,12h and 24h of hypoxia, the obvious degree of hypoxia effect is 6h <12h <24h. It is concluded that three polypeptides T1, T2 and T3 are non-toxic to RSC96 cell and possess anoxia resisting or hypoxia resisting effect.
(II) animal experiments:
BALB/C male mice were randomized into 5 groups (12 per group) each, a sedentary control group, a model control group, a milk residue polypeptide low dose group (25 mg/kg.d-1), a medium dose group (50 mg/kg.d-1), a high dose group (100 mg/kg.d-1), and fed once daily for 7 weeks. Each group was free to adapt to swim with weight at week 1, and the model control group and peptide dose group continued to perform weight-bearing swimming training for the next 6 weeks, referred to as excessive exercise. Finally, all mice were enrolled in voluntary activity for 10 minutes and voluntary activity data were recorded. The specific grouping and handling methods are shown in Table 1 below and the results are shown in Table 2.
Table 1 shows the method for treating the protection effect of the milk residue polypeptide on the fatigued mice
Grouping Number of examples Treatment method
Sedentary control group 12 Sodium carboxymethyl cellulose for stomach irrigation
Model control group 12 Training of swimming by using resina draconis and lavage of sodium carboxymethyl cellulose
Low dose set of resina Draconis 12 Gastric lavage milk residue polypeptide for training of swimming
Dose group in resina Draconis 12 Gastric lavage milk residue polypeptide for training of swimming
High dose set of resina Draconis 12 Gastric lavage milk residue polypeptide for training of swimming
TABLE 2 Effect of milk residue Polypeptides on voluntary locomotion in tired mice
Note that: comparison with sedentary control group + P≤0.05， ++ P is less than or equal to 0.01; p was less than or equal to 0.05 compared to the model control group; t mixing: and (3) performing low-temperature freeze drying on the enzymolysis liquid obtained by enzymolysis of milk residues to obtain a product (polypeptide).
As can be seen from Table 2, compared with the sedentary control group, the total distance, average speed and activity time of the model control group are obviously reduced (P is less than or equal to 0.05 or P is less than or equal to 0.01), and the rest time is obviously increased (P is less than or equal to 0.05); compared with the model control group, the rest time of the low, medium and high dosage groups of the polypeptides T mixed, T1, T2 and T3 is obviously reduced (P is less than or equal to 0.05), the total distance is obviously increased (P is less than or equal to 0.05). Conclusion: the milk residue polypeptide T mixture, T1, T2 and T3 have anti-fatigue effect on BALB/C mice, and T3 is optimal.
(III) human clinical trial
Experimental grouping: the method is divided into a plateau hypoxia control group, a ginseng rhodiola rosea capsule group, a plateau hypoxia experimental group and a normoxic control group.
Normoxic control group: recruiting 100 healthy volunteers, namely the original residents in the Hunan province, wherein the ratio of men to women is 1:1, the ages are 40-45 years, the life of the volunteers in the Hunan long sand urban area is according to usual life habits during the experiment period, and the physical and physiological states of the volunteers are detected after 1 week of the experiment;
plateau hypoxia control group: for the same group of volunteers in the normoxic control group, after the normoxic control experiment is finished, transferring personnel to the Tibetan Lasa urban area, and detecting the physical and physiological states of the volunteers according to usual living habits during the experiment for 1 week;
plateau hypoxia experimental group: taking the same batch of volunteers as the plateau hypoxia control group, after the plateau hypoxia control experiment is finished, feeding 20 volunteers with 7.0g of ginseng rhodiola root capsules per day according to 60kg of body weight during the experiment period, and detecting the physical and physiological states of the volunteers after the experiment for 1 week; taking 0.4g of anti-deficiency (low) freeze-dried powder every day for 20 volunteers according to the weight of 60kg of human body, and testing the physical and physiological states of the volunteers for 1 week; taking 0.4g T1 freeze-dried powder for 20 volunteers according to the weight of 60kg of human body every day, and testing the physical and physiological states of the volunteers for 1 week; taking 0.4g T2 freeze-dried powder for 20 volunteers according to the weight of 60kg of human body every day, and testing the physical and physiological states of the volunteers for 1 week; 20 volunteers were dosed with 0.4g of T3 lyophilized powder per day according to a human body weight of 60kg, and the physical and physiological status of the volunteers were examined for 1 week of the experiment. The results are shown in tables 3-4.
Table 3 clinical effects of milk residue polypeptide on male anti-hypoxia or anti-fatigue
Note that: p was less than or equal to 0.05 compared to the plateau hypoxia control group.
As can be seen from Table 3, the serum MDA of men in the anoxic control group is significantly increased (P.ltoreq.0.05 or P.ltoreq.0.01) and the SOD is significantly decreased (P.ltoreq.0.05) compared with the normoxic control group; compared with the anoxic control group, the ginseng rhodiola rosea capsule group and the polypeptide T mixed, T1, T2 and T3 groups have obviously reduced serum MDA (P is less than or equal to 0.05) and obviously increased SOD (P is less than or equal to 0.05). Conclusion: the Ginseng radix and radix Rhodiolae capsule and polypeptide T mixture, T1, T2, and T3 have anti-anoxia or hypoxia and anti-fatigue effects on male.
Table 4 clinical effects of milk residue polypeptide on female anti-hypoxia or anti-fatigue
Note that: p was less than or equal to 0.05 compared to the plateau hypoxia control group.
As can be seen from Table 4, compared with the normoxic control group, the serum MDA of the women in the anoxic control group is significantly increased (P is less than or equal to 0.05 or P is less than or equal to 0.01), and the SOD is significantly decreased (P is less than or equal to 0.05); compared with the anoxic control group, the ginseng rhodiola rosea capsule group and the polypeptide T mixed, T1, T2 and T3 groups have obviously reduced serum MDA (P is less than or equal to 0.05) and obviously increased SOD (P is less than or equal to 0.05). Conclusion: the Ginseng radix and radix Rhodiolae capsule and polypeptide T mixture, T1, T2, and T3 have anti-anoxia or hypoxia and anti-fatigue effects for female.
Cell test results show that the milk slag polypeptide and three polypeptides T1, T2 and T3 have better protection effect on nerve cells under the condition of hypoxia.
Animal experiments show that the milk residue polypeptides T, T1, T2 and T3 can obviously improve the autonomous activity of the fatigued mice.
The human clinical experiments show that the milk residue polypeptide and three polypeptides T1, T2 and T3 thereof can obviously reduce the symptoms of dizziness, headache, fatigue, weakness, insomnia and the like of a human body under the condition of altitude hypoxia, reduce the rising rate of the serum MDA content and the falling rate of the SOD content of the human body, and have better effect than the ginseng rhodiola root capsule. The yak milk residue anti-hypoxia or hypoxia anti-fatigue peptide prepared by the invention can be applied to health care products, can improve the conditions of human body, such as diseases, mental stress and plateau hypoxia environment, and has the advantages of small side effect, definite curative effect and high cost performance.
The above embodiments are merely preferred embodiments of the present invention, and should not be construed as limiting the present invention, and the embodiments and features in the embodiments in the present application may be arbitrarily combined with each other without collision. The protection scope of the present invention is defined by the claims, and the protection scope includes equivalent alternatives to the technical features of the claims. I.e., equivalent replacement modifications within the scope of this invention are also within the scope of the invention.
SEQUENCE LISTING
<110> university of forestry science and technology in south China
<120> a method for expressing an anti-hypoxic peptide
<130> 2022
<160> 3
<170> PatentIn version 3.5
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<211> 9
<212> PRT
<213> Bos mutus
<400> 2
Tyr Pro Phe Pro Gly Pro Ile Pro Asn
1 5
<210> 3
<211> 11
<212> PRT
<213> Bos mutus
<400> 3
Leu Val Tyr Pro Phe Pro Gly Pro Ile Pro Asn
1 5 10

Claims (8)

1. The application of the yak milk residue peptide in preparing the anti-hypoxia medicament is characterized in that the yak milk residue peptide is T1, T2 or T3, the amino acid sequence of T1 is shown as SEQ ID NO.1, the amino acid sequence of T2 is shown as SEQ ID NO.2, and the amino acid sequence of T3 is shown as SEQ ID NO. 3.
2. The application of the yak milk residue peptide in preparing the anti-hypoxia medicament according to claim 1, wherein the yak milk residue peptide is prepared from yak milk in the region with the Tibetan plateau altitude of 2800-5500 m and the low oxygen concentration of 10-14%.
3. The application of the yak milk residue peptide in preparing the anti-hypoxia medicament according to claim 1, wherein the preparation method of the yak milk residue peptide comprises the following steps of,
1) The yak milk residue sampling method comprises the following steps: freeze-dried yak milk residue powder or milk residue blocks by natural wind;
2) Pulverizing the yak milk residue powder or the milk residue blocks obtained in the step 1) to obtain yak milk residue fine powder;
3) Preparing yak milk residue peptide: adding distilled water and enzyme into the yak milk residue fine powder obtained in the step 2), carrying out ultrasonic crushing and enzymolysis, and carrying out freeze drying on the obtained zymolyte to obtain the yak milk residue peptide.
4. The use of yak milk residue peptide according to claim 3 for preparing an anti-hypoxia medicament, wherein the drying temperature in step 1) is: -10 o C~-25 o And C, air-drying, wherein the average air speed is 1.5-3m/s.
5. The use of yak milk residue peptide according to claim 3 for preparing anti-hypoxia drugs, wherein the crushing conditions of step 2) are: 11500-12500rpm, and shearing for 4.5-5.5min.
6. The use of the yak milk residue peptide in preparing anti-hypoxia drugs according to claim 3, wherein the step 3) is characterized in that the yak milk residue fine powder is prepared by the following steps: distilled water is added into 2-4 g/ml, and the ratio of the enzyme addition amount to the yak milk residue fine powder mass is 1: (50-150), ultrasonic crushing, and carrying out enzymolysis for 2-4 hours at the temperature of 32-42 ℃ in a water bath.
7. The use of yak milk residue peptide according to claim 6 for preparing anti-hypoxia drugs, wherein the enzymes of step 3) are pepsin and trypsin, and the efficacy of pepsin is 1: (25000-35000) the enzyme activity of trypsin is more than 250USP u/mg.
8. The application of the yak milk residue peptide in preparing the anti-hypoxia drugs according to claim 3, wherein the step 3) is carried out for 2-4 hours each time, after each time of enzymolysis is finished, centrifugation is carried out at 3000-5000rpm, supernatant fluid is reserved, milk residue sediment is continuously added with enzymolysis liquid for next enzymolysis, and finally all the supernatant fluid is collected for analysis, and each time of enzymolysis is carried out for 1-3 times.
CN202210453879.XA 2022-04-27 2022-04-27 Anti-hypoxia or hypoxia anti-fatigue peptide prepared from yak milk residues, and preparation method and application thereof Active CN114989247B (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105142658A (en) * 2013-03-15 2015-12-09 Mjn美国控股有限责任公司 Reducing proinflammatory response
CN112094880A (en) * 2020-08-30 2020-12-18 中南林业科技大学 Preparation method of yak milk residue polypeptide with antioxidant activity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105142658A (en) * 2013-03-15 2015-12-09 Mjn美国控股有限责任公司 Reducing proinflammatory response
CN112094880A (en) * 2020-08-30 2020-12-18 中南林业科技大学 Preparation method of yak milk residue polypeptide with antioxidant activity

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
"Ex vivo digestion of bovine milk with genetic variants A1 and A2 of beta-casein and identification of bioactive peptides";Asledottir,Tora等;Norwegian University of Life Sciences;表9-11 *
"Peptides Isolated from Yak Milk Residue Exert Antioxidant Effects through Nrf2 Signal Pathway";Feiyan Yang等;Oxidative Medicine and Cellular Longevity;摘要、材料与方法、表1 *

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