CN107252001B - A kind of method and application of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent - Google Patents
A kind of method and application of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent Download PDFInfo
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- CN107252001B CN107252001B CN201710448634.7A CN201710448634A CN107252001B CN 107252001 B CN107252001 B CN 107252001B CN 201710448634 A CN201710448634 A CN 201710448634A CN 107252001 B CN107252001 B CN 107252001B
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- Life Sciences & Earth Sciences (AREA)
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- Polymers & Plastics (AREA)
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- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Birds (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
The invention discloses the methods and application of a kind of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent.Present invention separation and Extraction epidermal growth factor EGF from the chicken embryo that fast-growth is developed, method is to make EGF band positive electricity according to the pH value for adjusting homogenate, EGF is adsorbed using benzoic acid not soluble in water, separate it from homogenate, acetone is recycled not dissolve the property of EGF, the principle design that EGF is parsed from benzoic acid, prepares a kind of nutrition renovation agent of the promotion animal intestinal mucosa reparation of EGF of source containing chicken.Animal intestinal mucosa EGF nutrition renovation agent of the invention adds 0.05-0.3 kilograms in livestock and poultry complete feed per ton, can significantly improve animal intestinal mucosa structure function, improves livestock and poultry feed intake and growth performance, significantly reduces diarrhea of weaned piglets rate.
Description
Technical field
The present invention relates to the methods and application of a kind of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent.
Background technique
Animal intestinal tract is not only the place of nutrient digestion absorption, is also important immune organ, safeguards normal intestines
Road function is most important to animal growth.Animal lacks in nutrient, infection, stress and when pathology, can lead
The damage of enteron aisle structure and function, bacteria and endotoxin translocation and gut muco-membranous barrier dysfunction etc. are caused, animal is seriously affected
Health and production performance bring weight huge economic loss to breeding production.Epidermal growth factor (epidermal growth
Factor, EGF) belong to somatomedin family member, there is extensive bioactivity, have and promote gastrointestinal tissue's growth and hair
It educates, repairs impaired mucous membrane tissue, adjust the various enzymatic activitys of enteron aisle, improve the important function such as nutrient digestion absorption.External source
Supply EGF is the important channel for improving animal intestinal tract structure and function, repairing damage enteron aisle, promoting animal health growth, and EGF makees
There is very wide application prospect on Animal husbandry production for novel animal intestinal mucosa nutrition regulator.
EGF is the single chain polypeptide being made of 53 amino acid, and secondary structure is fine and close chondritic, special space
Structure makes EGF have higher stability, has tolerance to acid, heat, can resist trypsase, pepsin and chymotrypsin
Digestion.EGF has many biological actions, can promote intracellular DNA, RNA and protein synthesis；Stimulate Various Tissues cell point
It splits, be proliferated and break up；Gastric acid secretion inhibiting promotes ulcer healing and epithelial tissue regeneration；Increase substance in vivo transhipment and metabolism；
Promote extracellular matrix and some substances of body (tartaric acid, prostaglandin) synthesis, has and adjust sperm, the development of ovary and reproduction
The effect of function, EGF is the important growth factor for promoting new born animal stomach, intestinal growth, but the limited source EGF, valuableness
Price restrict it in the extensive use in herding field.
Summary of the invention
The object of the present invention is to provide a kind of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent method,
It is characterized in that its step are as follows:
1) 15-20 age in days hatching chicken embryo is taken, decladding is added 0.05mol/L acetic acid solution high-speed homogenization 10-30 minutes,
1500 revs/min, wherein decladding chicken embryo and acetic acid solution mass ratio are 1:5-10, are centrifuged 20-30 minutes, take supernatant；
2) precipitating continues plus 0.05mol/L acetic acid solution is homogenized, 1500 revs/min of centrifugations, wherein precipitating and acetic acid solution
Mass ratio is 1:5-10, is centrifuged 20-30 minutes, and supernatant twice is merged, and removes supernatant surface fat,
3) supernatant adds sodium benzoate, is sufficiently stirred, and dissolves it all, with the NH of 3mol/L4OH solution tune pH is extremely
5.5-6.5 stirs half an hour, stands, is filtered under diminished pressure, and takes out precipitating；
4) after drying to be precipitated, add pre- cold acetone, stirring forms suspension, 4-12h is stood at 4 DEG C, then has albumen sample wadding
Shape precipitating, is filtered under diminished pressure；Precipitating is used into acetone washing again, zeolite powder is added, being precipitated after washing with the mass ratio of zeolite powder is 1:
5-10 is stirred 10-20 minutes, and animal intestinal mucosa EGF nutrition renovation agent is made in 50 DEG C of drying and crushings.
Preferably, the chicken embryo refers to white plumage AA chicken or the Sanhuang chicken chicken embryo of the healthy development of 15-20 age in days.
EGF of the present invention is a single chain polypeptide being made of 53 amino acid, and secondary structure is the spherical of densification
Structure.
Preferably, the step 1) and the concentration of the acetic acid solution in step 2) are 0.05mol/L, and temperature is 4 DEG C.
Preferably, the sodium benzoate additional amount is often to go up clear liquid 20-30g is added.
Preferably, after pre- cold acetone being added in the step 4), the mass concentration being deposited in pre-cooling acetone soln is
0.04-0.1g/ml, pre-cooling acetone temperature are 4 DEG C.
Preferably, the zeolite powder pH is between 7-7.5, water content 3.0-8.0%, and suction ammonia amount is 120-140 mmoles
That/100g, mesh number are 30-60 mesh.
The invention also discloses a kind of animal intestinal mucosa EGF nutrition renovation agents of the method preparation in livestock and poultry complete feed
In application, specifically: in livestock and poultry complete feed per ton add the 0.05-0.3 kilograms of animal intestinal mucosa EGF nutrition reparation
Agent.
Further, the 0.1-0.3 kilograms of animal intestinal mucosa is added in 21-28 age in days weanling pig complete feed per ton
EGF nutrition renovation agent adds the 0.05-0.2 kilograms of animal intestinal mucosa EGF nutrition renovation agent in sow complete feed per ton,
The 0.05-0.1 kilograms of animal intestinal mucosa EGF nutrition renovation agent is added in broiler chicken complete feed per ton.
The animal intestinal mucosa EGF nutrition renovation agent preparation method of the present invention is made due to using by adjusting solution ph
Chicken embryo is homogenized EGF band positive electricity, adsorbs EGF using benzoic acid not soluble in water, separates it from supernatant, recycles
Acetone does not dissolve the property of EGF, and the principle that EGF is parsed from benzoic acid can make chicken embryo EGF separation and Extraction obtain yield big
In 85%, EGF coarse powder yield be 0.1-0.2mg/g chicken embryo, it efficiently solve EGF limited source in existing feed industry,
Expensive problem.
Animal intestinal mucosa EGF nutrition renovation agent of the invention can make piglet average daily gain improve 30%-40%, material weight
Than reducing 10%-20%, diarrhea rate declines 80%-85%, and jejunum villi height improves 70%-90%, Crypt depth decline
20%-30%, duodenum enterokinase activity improve 50%-55%, and the activity of Jejunal mucosa lactase improves 35%-40%,
Sucrase active improves 40%-60%；Test group broiler chicken daily gain is set to improve 8-12%, feed intake improves 6-10%, and 12 refer to
Intestinal villus length improves 10-20%, and Duodenal villi length/Crypt depth improves 20-30%；Ileum villus length/crypts
Depth improves 10-18%, and 40- has been respectively increased in duodenum trypsase, amylase, lipase and total protein hydrolytic enzyme activities
55%, 80-100%, 30-45% and 25-35%.Animal intestinal mucosa EGF nutrition renovation agent of the invention has protection and promotes
The effect of mucosal epithelial cells growth, improves livestock and poultry feed intake and growth performance, significantly reduces the work of Diarrhea after Piglets " Weaning rate
With.
Specific embodiment
Present invention combination following instance is further described.
Embodiment 1
100 15 ages in days are taken to hatch healthy chicken embryo, decladding, 0.05mol/L acetic acid is added in the ratio of 1:6 (g/g) in weighing
(4 DEG C), high-speed homogenization 30 minutes, 1500 revs/min were centrifuged 20 minutes, took supernatant；Precipitating continues plus the acetic acid of 0.05mol/L
(4 DEG C) are homogenized 15 minutes, and 1500 revs/min are centrifuged 20 minutes, take supernatant, merge supernatant twice, remove supernatant surface
Fat is added sodium benzoate (25g/L) in supernatant, is sufficiently stirred, and dissolves its all, adds the NH of 3mol/L4OH tune PH is extremely
6.0, half an hour is stirred, stands, is filtered under diminished pressure, precipitating is taken out and adds pre- cold acetone (4 DEG C of 0.05g/ml acetone) after precipitating is dry,
Stirring forms suspension, stands 12h at 4 DEG C, albumen sample flocculent deposit occurs, be filtered under diminished pressure, and precipitating is used acetone washing 3 again
Secondary, zeolite powder (30 mesh) is added with the ratio of 1:5 in the precipitating left, is stirred 10 minutes, animal is made in 50 DEG C of drying and crushings
Intestinal mucosa EGF nutrition renovation agent.
Embodiment 2
25 age in days weaned piglets start, and control group fed compares daily ration (formula is shown in Table 1).Test group complete feed is per ton
200 grams of animal intestinal mucosa EGF nutrition renovation agent is added on the basis of control group daily ration, feeds above-mentioned feed to 46 ages in days, measurement is young
Pig growth performance.
Table 1 tests diet composition and nutritive index
Test result shows that compared with the control group test group piglet average daily gain improves 31% (P < 0.05), feed-weight ratio
12% (P < 0.05) is reduced, diarrhea rate declines 85% (P < 0.05), and jejunum villi height improves 72% (P < 0.05), Crypt depth
Decline 21% (P < 0.05), duodenum enterokinase activity improves 58% (P < 0.05), and the activity of jejunal mucous membrane lactase improves
39% (P < 0.05), sucrase active improve 42% (P < 0.05).
Embodiment 3
21 age in days healthy AA broiler chicken, control group fed compare daily ration (formula is shown in Table 2), and test group complete feed is per ton right
According to 100 grams of animal intestinal mucosa EGF nutrition renovation agent is added on the basis of group daily ration, above-mentioned feed is fed to 35 ages in days, measures broiler chicken
Growth performance.
Table 2 tests diet composition and nutritive index
Test result shows compared with the control group, test group broiler chicken daily gain and feed intake be respectively increased 9.17% (P <
0.05) and 7.74% (P < 0.05), Duodenal villi length improve 14.20% (P < 0.05), Duodenal villi length/hidden
Nest depth improves 25.53% (P < 0.01)；Ileum villus length/Crypt depth improves 13.63% (P < 0.05), duodenum pancreas
Protease, amylase, lipase and total protein hydrolytic enzyme activities be respectively increased 44.61% (P < 0.05), 98.34% (P <
0.01), 36.47% (P < 0.05) and 28.36% (P < 0.05).
Claims (9)
1. a kind of method of chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent, it is characterised in that its step are as follows:
1) 15-20 age in days hatching chicken embryo is taken, decladding adds 0.05mol/L acetic acid solution high-speed homogenization 10-30 minutes, 1500 turns/
Minute, wherein decladding chicken embryo and acetic acid solution mass ratio are 1:5-10, are centrifuged 20-30 minutes, take supernatant；
2) precipitating continues plus 0.05mol/L acetic acid solution is homogenized, 1500 revs/min of centrifugations, wherein precipitating and acetic acid solution quality
Than being centrifuged 20-30 minute for 1:5-10, merge supernatant twice, removal supernatant surface fat,
3) supernatant adds sodium benzoate, is sufficiently stirred, and dissolves it all, with the NH of 3mol/L4OH solution tune pH to 5.5-6.5,
Half an hour is stirred, stands, is filtered under diminished pressure, takes out precipitating；
4) after drying to be precipitated, add pre- cold acetone, stirring forms suspension, 4-12h is stood at 4 DEG C, then has albumen sample cotton-shaped heavy
It forms sediment, is filtered under diminished pressure；Precipitating to be used into acetone washing again, zeolite powder is added, being precipitated after washing with the mass ratio of zeolite powder is 1:5-10,
It is stirred 10-20 minutes, animal intestinal mucosa EGF nutrition renovation agent is made in 50 DEG C of drying and crushings.
2. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
It is, the chicken embryo refers to white plumage AA chicken or the Sanhuang chicken chicken embryo of the healthy development of 15-20 age in days.
3. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
It is that the EGF is a single chain polypeptide being made of 53 amino acid, secondary structure is fine and close chondritic.
4. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
The concentration for being the acetic acid solution is 0.05mol/L, and temperature is 4 DEG C.
5. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
It is that the sodium benzoate additional amount is often to go up clear liquid 20-30g is added.
6. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
It is after pre- cold acetone is added in the step 4), the mass concentration being deposited in pre-cooling acetone soln is 0.04-0.1g/ml,
It is 4 DEG C that acetone temperature, which is pre-chilled,.
7. the method for chicken embryo EGF separation and Extraction preparation animal intestinal mucosa nutrition renovation agent according to claim 1, feature
It is the zeolite powder pH between 7-7.5, water content 3.0-8.0%, suction ammonia amount is 120-140 mMs/100g, mesh number
For 30-60 mesh.
8. the animal intestinal mucosa EGF nutrition renovation agent that one kind is prepared method according to claim 1 is in livestock and poultry complete feed
Application, which is characterized in that in livestock and poultry complete feed per ton add the 0.05-0.3 kilograms of animal intestinal mucosa EGF nutrition reparation
Agent.
9. application according to claim 8, it is characterised in that: added in 21-28 age in days weanling pig complete feed per ton
The 0.1-0.3 kilograms of animal intestinal mucosa EGF nutrition renovation agent, in sow complete feed per ton described in 0.05-0.2 kilograms of addition
Animal intestinal mucosa EGF nutrition renovation agent, 0.05-0.1 kilograms of the addition animal intestinal mucosa EGF battalion in broiler chicken complete feed per ton
Support renovation agent.
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| CN112755177B (en) * | 2021-03-15 | 2022-03-18 | 浙江大学 | Oral liquid containing chick embryo EGF, preparation method and application |
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