CN106518960A - Method for preparing animal viscera micro-molecule polypeptide through complete physical method - Google Patents
Method for preparing animal viscera micro-molecule polypeptide through complete physical method Download PDFInfo
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- CN106518960A CN106518960A CN201610890699.2A CN201610890699A CN106518960A CN 106518960 A CN106518960 A CN 106518960A CN 201610890699 A CN201610890699 A CN 201610890699A CN 106518960 A CN106518960 A CN 106518960A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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Abstract
The invention provides a method for preparing animal viscera micro-molecule polypeptide through a complete physical method. The method for preparing animal viscera micro-molecule polypeptide through the complete physical method comprises the following steps of 1 animal viscera organ preprocessing, 2 animal viscera homogenizing, 3 freeze thawing, 4 low-temperature grinding, 5 superfine grinding, 6 low-temperature high-pressure protein grinding, 7 superfine separation and 8 freeze drying. The method for preparing animal viscera micro-molecule polypeptide through the complete physical method applies the complete physical method for preparing the animal viscera micro-molecule polypeptide, the influence caused by a chemical reagent on micro-molecule polypeptide active ingredients and the like is avoided, and the natural and efficient small peptide active substances are obtained on the basis of ensuring product safety.
Description
Technical field
The invention belongs to physics and food engineering technical field, more particularly to a kind of complete Physical preparation animal is dirty
The method of device micromolecule polypeptide.
Background technology
Since recent decades, though oneself has as many as hundreds of kind from detached biologically active peptide in various biologies, and it is at present, biological
Active peptide does not also have more consistent sorting technique.Biologically active peptide can be divided into exogenous and endogenous biological according to source to divide
Active peptide；Divide according to function and can be divided into physiologically active and food sense organ peptide.
Bioactive Peptides refer to what non-body was produced, the peptide with biologically active after being absorbed in the form of peptide
Class material, such as animal milk can provide various active skin, including milk-derived EGF (EGF), TGF
(TGF) etc..Endogenous bioactive peptide refers to that what the tissue of organism itself or organ produced has Physiological effect to itself
The peptide matters of effect, the main peptide hormone for including some important incretory secretions in vivo, such as growth hormone release swash
Element, thyroid-stimulating hormone, thymus gland skin, insulin etc.；Skin, such as bradykinin are swashed by the tissue produced in blood or tissue.
For a long time, people think that protein digested can only could be absorbed in the form of free amino acid always,
Utilization of the animal intestinal to protein is not limited to the form of free amino acid in fact, and major part is with 2～3 amino acid
The oligopeptides form of composition is absorbed.These peptide matters are the Molecularly Imprinted Polymers between protein and amino acid, can
Direct participation is ingested, digests, metabolic process, and its mechanism of absorption is better than protein and amino acid, and has amino acid and protein
Irreplaceable physiological function.It can be used as medicine, vaccine, diagnostic reagent, enzyme inhibitor and lead compound.Scientist
Study and be found that the active skin with the function such as immunological regulation, hypotensive, antiviral, antitumor, they are used for medical treatment, guarantor
The industries such as strong and food.
At present, the domestic technology for preparing animal viscera polypeptide mainly has following several：
1. biologic enzymolysis method；2. Hydrolyze method；3. physics prepares method.
Wherein：
1. biologic enzymolysis method
Using multiple protein enzyme (such as hydrolase) protein degradation, enter through very loaded down with trivial details Biochemical processes afterwards
The further isolated or purified of row.
Enzymatic isolation method requirement condition is high, and separation process is loaded down with trivial details tediously long, and intermediate product is excessive, is difficult to carry out pure separation, and,
As the continuation of enzyme is acted on, the inactivation of amount of activated material can be caused, and easily polluted.It is generally to be applied to the indoor system of experiment more
It is standby, and really it is used for the less of prepare with scale.Furthermore, using yielding poorly that enzymatic isolation method is produced, the rate of producing into<5%, product cost
It is high.
2. Hydrolyze method
Including alkaline process and acid system, cardinal principle is to control pH by acid, alkali process, makes protein denaturation, degraded or free,
It is finally reached separation, the purpose of purifying.
Hydrolyze method using it is more be acid system, however, being the damage that acid system or alkaline process can cause some effective ingredients
Lose, such as acid hydrolyzation can cause the loss of whole tryptophans, part serine and tyrosine, and working condition is poor, corrosivity is strong,
And produce a large amount of wastes；Alkaline process can make amino acid that racemization occurs, and make the big portion such as threonine, serine, arginine, cystine
Divide inactivation.The product yield of Hydrolyze method technology of preparing only has 2-3%.
3. physics prepares method
It is mainly used in the preparation of animal viscera freeze-dried powder and freeze-dried powder capsule, its technology is as follows：
Animal viscera pre-processes → shreds → freeze → low-temperature grinding → lyophilized → capsule/packaging
Physics prepares method and can not make the protein degradation of macromolecular structure, thus cannot be in separating animal's internal organs it is effective into
Part, can not carry out it is pure isolate and purify, therefore be only used for the production of coarse products as freeze-dried powder.The absorbability of its product
Difference, bioavailability are very low.
The content of the invention
This patent just, on the basis of physics preparation method, by the improvement to technology, can be prevented effectively from conventional physical
Problem present in method preparation, obtains the active ingredients such as animal viscera small molecular peptide, and then improves yield and the life of product
Thing utilization rate.
The method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide, comprises the steps：
Step one, animal viscera pretreatment：Clean dirt, the adipose tissue for separating and removing animal viscera and connective group
Knit, shred, clean haemolysis；
Step 2, animal viscera homogenate：Under aseptic condition, unnecessary distilled water is removed in control, the tissue and nothing by shredding, after haemolysis
Bacterium distilled water presses material-water ratio 30-60% (w/v) mixing, then uses historrhexis's homogenizer, is homogenized in small, broken bits；
Step 3, freeze thawing：Animal viscera homogenised tissue quick freeze 20-80h, Ran Houyu is made under the conditions of -20 to -70 DEG C
15-35 DEG C of water-bath is thawed；
Step 4, low-temperature grinding：Under the conditions of 0-10 DEG C, homogenate organs and tissues are crushed with low-temperature and high-speed refiner；
Step 5, ultramicro grinding：The animal viscera tissue fluid of low-temperature grinding presses material-water ratio 30-60% with sterile distilled water
(w/v) mix, be vortexed with low-temperature and high-speed and crush ultramicro grinding；
Step 6, cryogenic high pressure albumen are crushed：Under the conditions of 0-10 DEG C, with super-pressure continuous flow cell disintegrating machine, will be low
The organs and tissues that temperature is crushed carry out cell membrane and crush；
Step 7, ultra-fine separation：Target substance is intercepted using ultrafiltration and molecular sieve；
Step 8, freeze-drying：- 20 to -90 DEG C of low temperature quick freezes, are dried.
In a kind of preferred embodiment of method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide,
Clean haemolysis process to be specially in the step one：
Step one, by the animal viscera for shredding tissue with 1% bromogeramine solution soak 15-20min, sterile purified water
Rinse three times；
Step 2, with 75% alcohol-pickled 15-20min, sterile purified water rinses three times repeatedly；
Step 3, by material-water ratio 1:10-1:5 (w/v) add deionized water to carry out haemolysis, and the hemolysis time is 10-48 hours.
In a kind of preferred embodiment of method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide,
In the step 5, low-temperature and high-speed vortex grinder works under 18000 revs/min of condition of work at 2 DEG C -10 DEG C.
In a kind of preferred embodiment of method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide,
The step 5 is repeated three to five crushing.
In a kind of preferred embodiment of method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide,
The step 6 is repeated three to five crushing.
The complete Physical of the present invention prepares the method for animal viscera micromolecule polypeptide and has the advantages that：
The complete Physical of the present invention prepares the method for animal viscera micromolecule polypeptide and establishes one kind and effectively can prepare
The physics technology of animal viscera micromolecule polypeptide.This technology can simply, efficiently improve the utilization rate of animal viscera, while protecting
Card active ingredient micromolecule polypeptide, amino acid equal size and activity.The technology is that a kind of complete Physical prepares micromolecule polypeptide
Technology；The technical matters is simple, convenient, easy to operate, it is adaptable to industrialization production；The application of the technology greatly can be opened
Send out the using value of animal viscera；The technology can ensure the activity of micromolecule polypeptide, it is to avoid the inactivation of primary amino acid.
This technology patent system prepares animal viscera micromolecule polypeptide using complete Physical, it is to avoid chemical reagent is to little point
The impact of sub- polypeptide active composition etc., on the basis of product safety is ensured, obtains the small peptide active material of efficiency natural.This product
With immunological regulation, nutrition and anti-oxidant as primary efficacy, there is wide application at aspects such as prevention, treatment and health cares.This is special
Sharp technology can rationally utilize animal viscera resource, improve the effective rate of utilization of animal viscera, and then improve the additional of animal viscera
Value, improves the productive value of animal.
Tied up under 2 DEG C of -10 DEG C of cryogenic conditions using ultramicro grinding, high speed vortex pulverizer produces mechanical shearing and ultrasonic wave
2 kinds of functions are crushed, makes animal viscera become molecular level size；Crushed using cryogenic high pressure albumen, by the internal organs group of low-temperature grinding
Knit carry out cell membrane crush, reach cytoclastic effect, at the same ensure intracellular matter activity and performance.
Description of the drawings
For the technical scheme being illustrated more clearly that in the embodiment of the present invention, below will be to making needed for embodiment description
Accompanying drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the present invention, for
For those of ordinary skill in the art, on the premise of not paying creative work, can be obtaining other according to these accompanying drawings
Accompanying drawing, wherein：
Fig. 1 is a kind of embodiment of the method that the complete Physical that the present invention is provided prepares animal viscera micromolecule polypeptide
Process chart；
Fig. 2 is that the cleaning haemolysis of the method that the complete Physical shown in Fig. 1 prepares animal viscera micromolecule polypeptide is a kind of real
Apply the process chart of example.
Specific embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the present invention, rather than the embodiment of whole.It is based on
Embodiment in the present invention, it is all other that those of ordinary skill in the art are obtained under the premise of creative work is not made
Embodiment, belongs to the scope of protection of the invention.
Fig. 1 is referred to, is the preparation of the method that the complete Physical for providing of the invention prepares animal viscera micromolecule polypeptide
A kind of process chart of embodiment of method.
Method S1 that the complete Physical prepares animal viscera micromolecule polypeptide comprises the steps：
Step S11, animal viscera pretreatment：Clean dirt, the adipose tissue for separating and removing animal viscera and connective group
Knit, shred, clean haemolysis；
Step S12, animal viscera homogenate：Under aseptic condition, unnecessary distilled water is removed in control, the tissue by shredding, after haemolysis with
Sterile distilled water presses material-water ratio 30-60% (w/v) mixing, then uses historrhexis's homogenizer, is homogenized in small, broken bits；
Step S13, freeze thawing：Animal viscera homogenised tissue quick freeze 20-80h, Ran Houyu is made under the conditions of -20 to -70 DEG C
15-35 DEG C of water-bath is thawed；
Step S14, low-temperature grinding：Under the conditions of 0-10 DEG C, homogenate organs and tissues are crushed with low-temperature and high-speed refiner；
Step S15, ultramicro grinding：The animal viscera tissue fluid of low-temperature grinding presses material-water ratio 30-60% with sterile distilled water
(w/v) mix, be vortexed with low-temperature and high-speed and crush ultramicro grinding；
Specifically, low-temperature and high-speed vortex grinder is worked under 18000 revs/min of condition of work, is entered repeatedly at 2 DEG C -10 DEG C
Three to five crushing of row；
Step S16, cryogenic high pressure albumen are crushed：Under the conditions of 0-10 DEG C, with super-pressure continuous flow cell disintegrating machine, will be low
The organs and tissues that temperature is crushed carry out cell membrane and crush；
Specifically, three to five crushing are repeated.
Step S17, ultra-fine separation：Target substance is intercepted using ultrafiltration and molecular sieve；
Step S18, freeze-drying：- 20 to -90 DEG C of low temperature quick freezes, are dried.
Referring to Fig. 2, Fig. 2 is the method that the complete Physical shown in Fig. 1 prepares animal viscera micromolecule polypeptide
A kind of process chart of embodiment of cleaning haemolysis.
Clean haemolysis process to be specially in step S11：
Step S111, by the animal viscera for shredding tissue with 1% bromogeramine solution soak 15-20min, aseptic distillation
Water is rinsed three times；
Step S112, with 75% alcohol-pickled 15-20min, deionized water rinses three times repeatedly；
Step S113, by material-water ratio 1:10-1:5 (w/v) add deionized water to carry out haemolysis, and the hemolysis time is that 10-48 is little
When.
The present invention complete Physical prepare method 1 of animal viscera micromolecule polypeptide and preparation method thereof have have as follows
Beneficial effect：
The complete Physical of the present invention prepares the method for animal viscera micromolecule polypeptide and establishes one kind and effectively can prepare
The physics technology of animal viscera micromolecule polypeptide.This technology can simply, efficiently improve the utilization rate of animal viscera, while protecting
Card active ingredient micromolecule polypeptide, amino acid equal size and activity.The technology is that a kind of complete Physical prepares micromolecule polypeptide
Technology；The technical matters is simple, convenient, easy to operate, it is adaptable to industrialization production；The application of the technology greatly can be opened
Send out the using value of animal viscera；The technology can ensure the activity of micromolecule polypeptide, it is to avoid the inactivation of primary amino acid.
This technology patent system prepares animal viscera micromolecule polypeptide using complete Physical, it is to avoid chemical reagent is to little point
The impact of sub- polypeptide active composition etc., on the basis of product safety is ensured, obtains the small peptide active material of efficiency natural.This product
With immunological regulation, nutrition and anti-oxidant as primary efficacy, there is wide application at aspects such as prevention, treatment and health cares.This is special
Sharp technology can rationally utilize animal viscera resource, improve the effective rate of utilization of animal viscera, and then improve the additional of animal viscera
Value, improves the productive value of animal.
Tied up under 2 DEG C of -10 DEG C of cryogenic conditions using ultramicro grinding, high speed vortex pulverizer produces mechanical shearing and ultrasonic wave
2 kinds of functions are crushed, makes animal viscera become molecular level size；Crushed using cryogenic high pressure albumen, by the internal organs group of low-temperature grinding
Knit carry out cell membrane crush, reach cytoclastic effect, at the same ensure intracellular matter activity and performance.
Embodiments of the invention are the foregoing is only, the scope of the claims of the present invention is not thereby limited, it is every using this
Equivalent structure or equivalent flow conversion that bright specification and accompanying drawing content are made, or directly or indirectly it is used in other related skills
Art field, is included within the scope of the present invention.
Claims (5)
1. a kind of method that complete Physical prepares animal viscera micromolecule polypeptide, it is characterised in that comprise the steps：
Step one, animal viscera pretreatment：Clean dirt, separate the adipose tissue with removal animal viscera and connective tissue, cut
Broken, cleaning haemolysis；
Step 2, animal viscera homogenate：Under aseptic condition, unnecessary distilled water is removed in control, the tissue and aseptic steaming by shredding, after haemolysis
Distilled water presses material-water ratio 30-60% (w/v) mixing, then uses historrhexis's homogenizer, is homogenized in small, broken bits；
Step 3, freeze thawing：Animal viscera homogenised tissue quick freeze 20-80h is made under the conditions of -20 to -70 DEG C, then in 15-35
DEG C water-bath is thawed；
Step 4, low-temperature grinding：Under the conditions of 0-10 DEG C, homogenate organs and tissues are crushed with low-temperature and high-speed refiner；
Step 5, ultramicro grinding：The animal viscera tissue fluid of low-temperature grinding presses material-water ratio 30-60% (w/v) with sterile distilled water
Mixing, is vortexed with low-temperature and high-speed and crushes ultramicro grinding；
Step 6, cryogenic high pressure albumen are crushed：Under the conditions of 0-10 DEG C, with super-pressure continuous flow cell disintegrating machine, by low temperature powder
Broken organs and tissues carry out cell membrane and crush；
Step 7, ultra-fine separation：Target substance is intercepted using ultrafiltration and molecular sieve；
Step 8, freeze-drying：- 20 to -90 DEG C of low temperature quick freezes, are dried.
2. the method that complete Physical according to claim 1 prepares animal viscera micromolecule polypeptide, it is characterised in that institute
Haemolysis process is cleaned in stating step one to be specially：
Step one, by the animal viscera for shredding tissue with 1% bromogeramine solution immersion 15-20min, sterile purified water flushing
Three times；
Step 2, with 75% alcohol-pickled 15-20min, sterile purified water rinses three times repeatedly；
Step 3, by material-water ratio 1:10-1:5 (w/v) add deionized water to carry out haemolysis, and the hemolysis time is 10-48 hours.
3. the method that complete Physical according to claim 1 prepares animal viscera micromolecule polypeptide, it is characterised in that institute
State in step 5, low-temperature and high-speed vortex grinder works under 18000 revs/min of condition of work at 2 DEG C -10 DEG C.
4. the method that complete Physical according to claim 1 prepares animal viscera micromolecule polypeptide, it is characterised in that institute
State step 5 and three to five crushing are repeated.
5. the method that complete Physical according to claim 1 prepares animal viscera micromolecule polypeptide, it is characterised in that institute
State step 6 and three to five crushing are repeated.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610890699.2A CN106518960A (en) | 2016-10-12 | 2016-10-12 | Method for preparing animal viscera micro-molecule polypeptide through complete physical method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610890699.2A CN106518960A (en) | 2016-10-12 | 2016-10-12 | Method for preparing animal viscera micro-molecule polypeptide through complete physical method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113072614A (en) * | 2021-04-13 | 2021-07-06 | 内蒙古蒙肽生物工程有限公司 | Equipment and method for preparing small molecular polypeptide of animal viscera |
| CN113287756A (en) * | 2021-05-26 | 2021-08-24 | 东莞十度生物科技有限公司 | Active Chinese medicine source of compound active polypeptide series and its prepn |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1524536A (en) * | 2003-02-24 | 2004-09-01 | 内蒙古神元生物工程股份有限公司 | Technology for producing sheep placenta biolobical preparations using low-temperature ultrasonic high-pressure egg albumen cracking method |
-
2016
- 2016-10-12 CN CN201610890699.2A patent/CN106518960A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1524536A (en) * | 2003-02-24 | 2004-09-01 | 内蒙古神元生物工程股份有限公司 | Technology for producing sheep placenta biolobical preparations using low-temperature ultrasonic high-pressure egg albumen cracking method |
Non-Patent Citations (1)
| Title |
|---|
| 赵云 等: "羊胎盘肽的提取及质量标准的初步研究", 《黑龙江医药科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113072614A (en) * | 2021-04-13 | 2021-07-06 | 内蒙古蒙肽生物工程有限公司 | Equipment and method for preparing small molecular polypeptide of animal viscera |
| CN113287756A (en) * | 2021-05-26 | 2021-08-24 | 东莞十度生物科技有限公司 | Active Chinese medicine source of compound active polypeptide series and its prepn |
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Application publication date: 20170322 |