CN114540447B - Animal myocardial zymolyte and preparation method thereof and application of animal myocardial zymolyte in preparation of products for improving myocardial function - Google Patents
Animal myocardial zymolyte and preparation method thereof and application of animal myocardial zymolyte in preparation of products for improving myocardial function Download PDFInfo
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- CN114540447B CN114540447B CN202210065629.9A CN202210065629A CN114540447B CN 114540447 B CN114540447 B CN 114540447B CN 202210065629 A CN202210065629 A CN 202210065629A CN 114540447 B CN114540447 B CN 114540447B
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/20—Meat products; Meat meal; Preparation or treatment thereof from offal, e.g. rinds, skins, marrow, tripes, feet, ears or snouts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
- A23L13/48—Addition of, or treatment with, enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
- A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/34—Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Cardiology (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Developmental Biology & Embryology (AREA)
- Heart & Thoracic Surgery (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Hospice & Palliative Care (AREA)
- Vascular Medicine (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses an animal myocardial zymolyte, a preparation method thereof and application thereof in preparing products for improving myocardial function. The preparation method of the zymolyte comprises the following steps: an animal heart minced meat, repeatedly frozen and thawed; the meat emulsion is subjected to heat treatment to inactivate enzyme, water is added to control the temperature to adjust the pH value, and protease is added to carry out enzymolysis for 4 to 10; and after enzymolysis, carrying out heat treatment on the reaction system to inactivate enzyme, centrifuging to collect supernatant, concentrating moderately, and then spray drying or freeze drying to obtain the animal myocardial enzymolysis product. The obtained zymolyte small molecule peptide has high ratio, can directly act on myocardial cells, and obviously improves the capability of resisting damage of the myocardial cells. The zymolyte obtained by the invention protects the heart of the mouse in a heart failure model of the doxorubicin of the mouse, has remarkable effect, reduces the damage of the doxorubicin to the heart of the mouse, and has good market development and application prospects.
Description
Technical Field
The invention belongs to the field of bioactive peptides, and in particular relates to animal myocardial zymolyte, a preparation method thereof and application thereof in preparing products for improving myocardial function.
Background
The heart is the most important organ in viscera, and provides power for blood circulation; the heart is a hollow muscle tissue, and the myocardium, which is the main component of the heart, serves as the outline of the heart's cavity. The heart muscle rhythmically contracts and expands to form the beating of the heart, which is the functional basis for the heart to exert its pumping action.
The irregular life and cardiovascular diseases can cause myocardial injury with different degrees, and the myocardial injury can be easily developed to the later stage to cause heart failure and death. Congenital heart disease, hypertension and coronary heart disease are high incidence people of heart failure.
In the study of myocardial protection, doxorubicin is used as a clinical medicine for treating cancers and inhibiting tumors, and can cause myocardial damage, and is used as a model medicine for myocardial damage by a large number of researchers. The chemical medicines always have or have light or heavy side effects, and the chemical medicines are required to be taken in compliance with medical advice, and the food-borne active substances have no toxic or side effects, can be taken as daily health care, reduce stress reaction caused by stimulation of cardiac muscle and improve injury effect, so that the development of the food-borne substances which have myocardial protection activity and are safe and nontoxic is significant.
Disclosure of Invention
The invention aims to provide an animal myocardial zymolyte, a preparation method thereof and application thereof in preparing products for improving myocardial function.
The aim of the invention is achieved by the following technical scheme:
a preparation method of animal myocardial zymolyte comprises the following steps:
(1) Taking animal hearts (animals are killed and slaughtered), removing surface fat, cutting into pieces and mincing into meat paste, freezing at-80 ℃ to-60 ℃ for 8-12 h, repeatedly thawing at 30-40 ℃ for 4-6 h, and repeatedly freezing and thawing for several times;
the animal is mammal, preferably cow, pig or sheep;
(2) Performing heat treatment on the meat emulsion obtained in the step (1) to inactivate enzyme, then adding water, controlling the temperature to be 45-60 ℃, adjusting the pH value of the system to be 7-9, adding protease with the mass of 1-4 per mill of the meat emulsion, and performing enzymolysis for 4-10 h;
the protease is more than one of alkaline protease, neutral protease or flavourzyme;
(3) After enzymolysis, carrying out heat treatment on the reaction system to inactivate enzyme, centrifuging to collect supernatant, concentrating moderately, and spray drying or freeze drying to obtain animal myocardial enzymolysis product;
the heat treatment in the steps (2) and (3) is preferably carried out at the temperature of 85-95 ℃ for 10-15 min;
the centrifugation in the step (3) is preferably carried out at the temperature of 4-10 ℃ for 10-20 min at 8000-10000 g;
concentrating in the step (3), preferably concentrating at 55 ℃ in vacuum until the solid content is 30-50%;
the spray drying in the step (3) is carried out, the air inlet temperature is preferably 160-210 ℃, and the air outlet temperature is preferably 70-120 ℃;
the temperature of the freeze drying in the step (3) is-50 to-40 ℃.
In the animal myocardial zymolyte prepared by the method, the number of polypeptides with the molecular weight of less than 500Da accounts for 59.11-79.67%, and the obtained zymolyte can obviously reduce the damage of doxorubicin to the heart of a mouse and can be used for preparing medicaments and health-care products for improving myocardial function.
The medicine can also contain more than one pharmaceutically acceptable carrier or auxiliary materials.
The auxiliary materials are preferably sustained release agents, excipients, fillers, adhesives, wetting agents, disintegrating agents, absorption promoters, adsorption carriers, surfactants or lubricants and the like.
The carrier is at least one of microcapsule, microsphere, nanoparticle and liposome.
Compared with the prior art, the invention has the following advantages and effects:
1. the obtained zymolyte small molecule peptide has high ratio, can directly act on myocardial cells, and obviously improves the capability of resisting damage of the myocardial cells.
2. The zymolyte obtained by the invention protects the heart of the mouse in a heart failure model of the doxorubicin of the mouse, has remarkable effect, reduces the damage of the doxorubicin to the heart of the mouse, and has good market development and application prospects.
3. The invention combines endogenous protease and commercial protease to carry out enzymolysis on food-borne protein, and the product is safe, has no toxic or side effect and has higher economic benefit; the preparation method can obtain the small molecular active peptide which is richer and easy to absorb, and the product has obvious efficacy.
4. The invention has simple process, low equipment requirement, strong operability and good application value.
Drawings
FIG. 1 shows the molecular weight distribution of polypeptides in the substrate.
FIG. 2 is the protective effect of the substrate on doxorubicin-induced cardiomyocyte injury.
FIG. 3 is the effect of the substrate on the short axis shortening rate of each group of doxorubicin-induced heart failure model mice.
FIG. 4 is the effect of the substrate on the ejection fraction of doxorubicin-induced heart failure model mice.
FIG. 5 is the effect of the substrate on left chamber volume of doxorubicin-induced heart failure model mice.
FIG. 6 is the effect of the enzyme substrate on lactate dehydrogenase activity in serum of mice with doxorubicin-induced heart failure model.
FIG. 7 is the effect of the substrate on the activity of creatine kinase isoenzyme in serum of mice with doxorubicin-induced heart failure model.
FIG. 8 is the effect of the substrate on superoxide dismutase activity in heart tissue of doxorubicin-induced heart failure model mice.
FIG. 9 is the effect of the enzymatic hydrolysate on the malondialdehyde content of lipid oxidative metabolite in heart tissue of a model of doxorubicin-induced heart failure.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but embodiments of the present invention are not limited thereto.
Example 1
A preparation method of animal myocardial zymolyte specifically comprises the following steps:
1) Pretreatment: after the beef heart is cleaned, the surface fat of the beef heart is removed, and the beef heart is cut into pieces and minced to be sufficiently fine.
2) Repeated freezing and thawing: sealing and subpackaging the minced meat into 200g, quick-freezing at-60 ℃ for 8h, thawing at 38 ℃ for 6h, and repeating the operation for 2 times.
3) Inactivating and inactivating enzyme: after the last thawing, the temperature is quickly raised to 95 ℃ and the enzyme is heated for 12min for the enzyme deactivation treatment.
4) Enzymolysis: respectively weighing and adding the mixture of the bovine heart blood and the distilled water according to the mass ratio of 1:1.5, heating to 55 ℃, stirring to uniformly distribute the meat emulsion in the water, and regulating the pH value of the system to 8. Adding neutral protease 2%of meat emulsion and flavourzyme 1%of meat emulsion, continuously stirring during enzymolysis process to make meat emulsion uniformly distributed in the system, uniformly contacted with protease, and performing enzymolysis for 6h.
5) Inactivating and inactivating enzyme: after the enzymolysis is completed to obtain an enzymolysis liquid, rapidly heating to 95 ℃ and heating for 10min to perform enzyme deactivation treatment, centrifuging 8000g at 4 ℃ for 15min to remove sediment, and collecting supernatant.
6) Concentrating: concentrating at 55deg.C under vacuum, concentrating the supernatant until the solid content is 40%, and concentrating.
7) Spray drying: supernatant with 40% solid content has inlet temperature of 160-210 deg.c and outlet temperature of 70-120 deg.c.
Example 2
A preparation method of animal myocardial zymolyte specifically comprises the following steps:
2) Repeated freezing and thawing: sealing and subpackaging the minced meat into 500g, quick-freezing at-80 ℃ for 10h, thawing at 35 ℃ for 5h, and repeating the operation for 3 times.
4) Enzymolysis: respectively weighing and adding the beef paste and distilled water according to the mass ratio of 1:1, heating to 55 ℃, stirring to uniformly distribute the beef paste in the water, and regulating the pH of the system to 8. Adding alkaline protease with mass of 2 per mill of meat emulsion, continuously stirring during enzymolysis process to ensure that the meat emulsion is uniformly distributed in the system, uniformly contacts with the protease, and carries out enzymolysis for 8 hours.
The other steps are the same as in example 1.
Example 3
A preparation method of animal myocardial zymolyte specifically comprises the following steps:
2) Repeated freezing and thawing: sealing and subpackaging the minced meat into 500g, quick-freezing at-80 ℃ for 12h, thawing at 37 ℃ for 4h, and repeating the operation for 3 times.
4) Enzymolysis: respectively weighing and adding the beef paste and distilled water according to the mass ratio of 1:3, heating to 50 ℃, stirring to uniformly distribute the beef paste in the water, and regulating the pH of the system to 7.5. Adding neutral protease with mass of 2 per mill of meat emulsion, continuously stirring during enzymolysis to ensure that the meat emulsion is uniformly distributed in the system, uniformly contacts with the protease, and carries out enzymolysis for 8 hours.
The other steps are the same as in example 1.
Comparative example 1
A preparation method of animal myocardial zymolyte specifically comprises the following steps:
prepared according to the method of example 1, except that: step 3) of example 1 was not performed to inactivate the enzyme, and commercial proteases (neutral protease and flavourzyme) were not added in the subsequent enzymatic hydrolysis process, the rest of the steps being the same as in example 1.
Comparative example 2
A preparation method of animal myocardial zymolyte specifically comprises the following steps:
prepared according to the method of example 1, except that: the protease used in the enzymolysis process is bromelain only, the enzyme adding amount is 2 per mill of the mass of meat emulsion, and the rest steps are the same as those of the example 1.
Effect verification example 1
Molecular weight distribution of the substrate
The molecular weight measurement method is as follows: determining the molecular weight distribution of the sample by gel chromatography; the standard peptide samples were: tripeptide GGG (relative molecular mass 189 Da), tetrapeptide GGYR (relative molecular mass 451 Da), aprotinin (relative molecular mass 6512 Da), cytochrome C (relative molecular mass 12384 Da), bovine serum albumin (relative molecular mass 66463 Da), and the linear equation fitted by the molecular weight of each standard and the retention time of each standard detected by high performance liquid chromatography is: y= -2.1128x+14.808 (R 2 =0.98), where y is the log molecular weight of the standard and x is the retention time.The molecular weight distribution of the samples was calculated by comparing the retention times of the samples. High performance liquid chromatography conditions: the chromatographic column TSK-GEL G2000 SWXL 7.8 mm. Times.300 mm, detection wavelength 220nm, flow rate 0.5mL/min, mobile phase 20% acetonitrile, 79.92% ultrapure water and 0.08% trifluoroacetic acid.
The experimental results are shown in fig. 1: the ratio of polypeptides less than 500Da in the examples is significantly higher than that in the comparative examples, and the ratios of polypeptides less than 500Da in the examples 1, 2 and 3 are 79.67%, 59.11% and 63.56%, respectively, wherein the ratio of the components less than 500Da in the example 1 is as high as 79.67%, which indicates that the use of the complex enzyme results in a significant increase in the small molecular weight (< 500 Da) polypeptides of the example 1; since comparative example 1 was free of added commercial protease, the small molecular components were significantly lower than the other groups, and the large molecular components were in high proportion, demonstrating that the use of a single endogenous protease released small molecular active peptide fragments in a limited manner, and the subsequent use of commercial proteases effectively acted on the long peptide sequences of the previous step, and released the small molecular components more efficiently.
Effect verification example 2
Protection effect of zymolyte on doxorubicin induced injury myocardial cells
The experimental method comprises the following steps:
cell viability: dissolving the zymolyte with DMEM high sugar culture medium to obtain 100 μg/mL, filtering, sterilizing, and inoculating rat myocardial cells H9c2 at a density of 5×10 5 Inoculating the sample group into a 96-well plate at a concentration of each mL/mL, after the inoculation is completed for about 24 hours, adding 200 mu L of prepared enzymolysis products with different concentrations into each well of the sample group to incubate, after 24 hours, stimulating injury of the sample group and the injury group by using 10 mu M doxorubicin with the same dosage, after 24 hours, adding 10% of cck8 reagent into a cell culture medium, and after incubation for 2 hours at 37 ℃ in a dark place, measuring a light absorption value at a wavelength of 450nm by using an enzyme-labeled instrument, and calculating the cell survival rate.
The experimental results are shown in fig. 2: the stimulation injury of doxorubicin can be seen to significantly reduce the survival rate of cells compared to the normal control, and the pre-protection effect of examples on cardiomyocytes is significantly stronger than that of comparative examples, with example 1 having the highest small molecule content having the best protection effect, whereas comparative example 1 without protease addition has little effect on the pre-protection of cardiomyocytes.
Effect verification example 3
Protection effect of zymolyte on doxorubicin-induced mice heart failure model
The experimental method comprises the following steps: after the C57BL/6J mice of about 8 weeks old are adaptively fed with common feed for one week, 10 groups are randomly grouped, the samples (zymolyte) are fed according to the dosage of 400mg/kg of the body weight of the mice per day in the group of the example 1 and the group of the example 2 for 4 weeks, the control group and the injury group are normally fed for 4 weeks, three groups (except the control group) after 4 weeks are uniformly subjected to intraperitoneal injection of doxorubicin solution for molding (20 mg/kg of the body weight of the mice), the mice are continuously cultured for about one week, ultrasonic heart is measured, and the materials are obtained after 12 hours of fasted.
Experimental results:
as shown in fig. 3-5, by analyzing the ultrasonic cardiac data of four groups of experimental mice, comparing the control group with the injury group, the short axis shortening rate, the ejection fraction and the left chamber volume of the mice treated by the doxorubicin are all obviously smaller than those of the normal group, and the doxorubicin is used as a chemical medicament, so that atrophy of partial tissues of the organism can be caused, the injury effect is brought to the organism, and the short axis shortening rate, the ejection fraction and the left chamber volume of the hearts of the mice which are pre-protected and then stimulated by the doxorubicin are obviously larger than those of the injury group, so that the injury brought to cardiac muscles by the doxorubicin is improved.
As shown in figures 6-7, the serum of the mice is collected, protease indexes in the serum of the mice are measured by a full-automatic serum analyzer, and compared with a control group and a damaged group, the lactate dehydrogenase and the creatine kinase isoenzyme in the serum of the mice after the doxorubicin injury are obviously increased, and the lactate dehydrogenase and the creatine kinase isoenzyme in the serum of the mice subjected to the pre-protection and the doxorubicin stimulation injury by the embodiment of the invention can be basically recovered to normal levels, so that the injury effect in the mice is improved to a certain extent.
As shown in fig. 8-9, collecting mouse myocardial tissue for on-ice lysis, centrifuging 12000g, collecting the supernatant, measuring the protein content by using a BCA protein assay, measuring the related indexes in the tissue by using a SOD and MDA kit built by south kyo, comparing the analysis data with a normal group to obtain the activity of superoxide dismutase (SOD) in the mouse myocardial tissue lysate after doxorubicin injury, which is obviously lower than that of a control group, wherein the activity of superoxide dismutase in the mouse myocardial tissue pre-protected by the embodiment of the invention is obviously higher than that of the injury group, and Malonaldehyde (MDA) is used as a lipid oxidation product, and the content of the superoxide dismutase in the myocardial tissue after doxorubicin injury is higher than that of the control group, and the content of the mouse myocardial tissue pre-protected by the embodiment of the invention is obviously lower than that of the injury group, so that the oxidative injury of doxorubicin to the myocardial tissue is improved to a certain extent.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (9)
1. The preparation method of the animal myocardial zymolyte is characterized by comprising the following steps:
(1) Taking animal hearts, removing surface fat of the animal hearts, cutting the animal hearts into pieces and mincing the meat mince, freezing the animal hearts at the temperature of between 80 ℃ below zero and 60 ℃ below zero for 8 to 12 hours, thawing the animal hearts at the temperature of between 30 and 40 ℃ for 4 to 6 hours, and repeating the freezing and thawing operations for a plurality of times;
(2) Performing heat treatment on the meat emulsion obtained in the step (1) to inactivate enzyme, then adding water, controlling the temperature to be 45-60 ℃, adjusting the pH value of the system to be 7-9, adding protease, and performing enzymolysis for 4-10 h;
the protease is a complex enzyme consisting of 2 per mill of neutral protease by mass of meat emulsion and 1 per mill of flavourzyme by mass of meat emulsion or 2 per mill of alkaline protease by mass of meat emulsion or 2 per mill of neutral protease by mass of meat emulsion;
(3) After enzymolysis, carrying out heat treatment on the reaction system to inactivate enzyme, centrifuging to collect supernatant, concentrating moderately, and spray drying or freeze drying to obtain animal myocardial enzymolysis product;
the animal in the step (1) is a mammal;
in the animal myocardial zymolyte, the number of the polypeptides with the molecular weight smaller than 500Da accounts for 59.11-79.67 percent.
2. The method of manufacturing according to claim 1, characterized in that: and (3) heat treatment, wherein the heat treatment is carried out at the temperature of 85-95 ℃ for 10-15 min.
3. The method of manufacturing according to claim 1, characterized in that: the centrifugation in the step (3) is carried out at the temperature of 4-10 ℃ for 10-20 min at 8000-10000 g.
4. The method of manufacturing according to claim 1, characterized in that: and (3) concentrating at 55 ℃ in vacuum until the solid content is 30-50%.
5. The method of manufacturing according to claim 1, characterized in that: and (3) spray drying, wherein the air inlet temperature is 160-210 ℃, and the air outlet temperature is 70-120 ℃.
6. The method of manufacturing according to claim 1, characterized in that: the temperature of the freeze drying in the step (3) is-50 to-40 ℃.
7. An animal myocardial enzymolysis product, which is characterized in that: is prepared by the method of any one of claims 1-6.
8. Use of an animal myocardial zymolyte according to claim 7 for the preparation of a medicament for improving myocardial function.
9. The use of the animal myocardial zymolyte of claim 7 in the preparation of a health care product.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1552733A (en) * | 2003-06-04 | 2004-12-08 | 大连珍奥药业有限公司 | Preparation of myocardium peptide |
| CN101130061A (en) * | 2007-07-24 | 2008-02-27 | 李立 | Segmented intestine targeted drug feeding preparation of myocardium protein polypeptide and method of producing the same |
| CN108607091A (en) * | 2016-12-12 | 2018-10-02 | 锡林郭勒盟肽好生物制品有限责任公司 | The formula of blood pressure lowering polypeptide draft solid beverage and myocardium peptide powder, preparation method thereof |
| CN110452948A (en) * | 2019-09-12 | 2019-11-15 | 北华大学 | Deer heart protein hydrolysate and its preparation method and application |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1552733A (en) * | 2003-06-04 | 2004-12-08 | 大连珍奥药业有限公司 | Preparation of myocardium peptide |
| CN101130061A (en) * | 2007-07-24 | 2008-02-27 | 李立 | Segmented intestine targeted drug feeding preparation of myocardium protein polypeptide and method of producing the same |
| CN108607091A (en) * | 2016-12-12 | 2018-10-02 | 锡林郭勒盟肽好生物制品有限责任公司 | The formula of blood pressure lowering polypeptide draft solid beverage and myocardium peptide powder, preparation method thereof |
| CN110452948A (en) * | 2019-09-12 | 2019-11-15 | 北华大学 | Deer heart protein hydrolysate and its preparation method and application |
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