CN107522782A - A kind of preparation method and applications of gradient collagen and collagen polypeptide molecule - Google Patents
A kind of preparation method and applications of gradient collagen and collagen polypeptide molecule Download PDFInfo
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- CN107522782A CN107522782A CN201710882480.2A CN201710882480A CN107522782A CN 107522782 A CN107522782 A CN 107522782A CN 201710882480 A CN201710882480 A CN 201710882480A CN 107522782 A CN107522782 A CN 107522782A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
- A23L2/52—Adding ingredients
- A23L2/66—Proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/80—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter by adsorption
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
- A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
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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
- 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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Abstract
" a kind of preparation method and applications of gradient collagen and collagen polypeptide molecule " of the invention, belong to the constant gradient molecule preparing technical field such as collagen, collagen polypeptide and amino acid oligomers, be related to preparation, production technology and its application of collagen and gradient collagen polypeptide, oligopeptides molecule etc..The claim protection gradient collagen of the present invention and the preparation method and applications of collagen polypeptide molecule.Including：Extract the pretreating process of collagen raw material；Extract the technique and technology of collagen macromolecular；By collagen macromolecular to 18KDa, to 3KDa, to 1.5KDa gradient collagen peptide molecule, and 18KDa, 3KDa, 1.5KDa collagen polypeptide molecule and oligomeric amino acid, the preparation method and operating procedure of amino acid, and gradient collagen molecules are in beauty industry, food additives, packaging material for food, health food, prevention disease and application in the beverage.
Description
Technical field
The invention belongs to collagen and collagen polypeptide gradient molecule preparing technical field, it is related to collagen and gradient glue
Preparation, production technology and its application of former polypeptide, oligopeptides molecule etc..
Background technology
Collagen (collagen) has an important physiological function in human body, including the growing of body, aging
Exchange and balance between the renewal of tissue, the reparation of injury tissue, the regulation of osmotic pressure, maintenance blood plasma and tissue fluid etc..
Collagen is a kind of protein of three-dimensional spiral structure, is the strong backing of dermal layer of the skin, and skin water is tender smooth to be had
Elasticity, and wrinkle, aging, slack and undisciplined, color spot etc. situation, caused by being all due to the holding and be lost in of collagen.
Collagen has 30 kinds or so, and common type is I type, II type, III type, IV type, V type and Ⅺ type.Collagen because
With biocompatibility, degradability and biological function, therefore in the fields such as food, medicine, organizational project, cosmetics extensively
Application.NTx albumen is distributed mainly on the tissue such as skin, tendon, rich in the hydroxylysine for thering is alanine to be combined with sugar type,
Molecular length about 300nm, diameter about 1.5nm, in bar-shaped, it is made up of three peptide chains, wherein having two α (I) chains, a α (II)
Chain, body function is acted on most strong；TypeⅡ Collagen is produced by cartilage cell；Type Ⅳ collagen albumen category basilar memebrane collagen egg
In vain, basilar memebrane is distributed mainly on, molecule is very tiny and is linearly distributed in organization internal, is the master for connecting epidermis and corium
Want key element；V Collagen Type VI egg is pericellular collagen, is distributed in around cell peripheral and NTx, may be in base
Work between film and connective tissue, largely exist in connective tissue.Collagen is human body natural's albumen, to skin surface
Protein molecule there is larger affinity, weaker antigenic, good biocompatibility and biodegradable security, can
Degraded absorbs, and adhesion strength is good.Applied to beauty, food, medicine, tonic.Early in 12nd century Bingen St.Hilde-gard
Just describe by the use of the cartilage soup of calf as medicine to treat arthralgia.It is a kind of preferable blood fat reducing food, can reduces
Blood triglyceride and cholesterol.Collagen is the key that calcium is pinned in skeleton, cartilaginous tissue.Or sclera it is main into
Point, the effect to eyes is also extremely important, if the synthesis of collagen is reduced in sclera, increase of degrading may result in myopia.
The present invention uses scientific and effective method, formed one kind prepare collagen and collagen polypeptide include macromolecular, in
The gradient molecule and collagen oligopeptide of equimolecular and small molecule and point out its practical case to the method and production technology of amino acid
Example.
The content of the invention
It is an object of the invention to provide the preparation method and applications of a kind of gradient collagen and collagen polypeptide molecule.
The preparation method and applications of gradient collagen and collagen polypeptide molecule provided by the invention, including following step
Suddenly：
1st, the pretreatment process method of collagen raw material is extracted；
2nd, the technique and technology of collagen macromolecular are extracted；
3rd, by the collagen gradient molecule of collagen macromolecular to 18KDa or so, and 18KDa or so collagen egg
White molecule preparation method and operating procedure；
4th, collagen molecules by 18KDa or so are prepared to the gradient collagen peptide molecule for being less than 3KDa, and 3KDa
Collagen polypeptide molecule operating procedure；
5th, prepare and 3KDa collagen polypeptide is less than to the collagen polypeptide of the relatively small molecule less than 1.5KDa by molecular weight, and
The preparation method and operating procedure of collagen oligopeptide, amino acid oligomers and amino acid less than 1.5KDa；
6th, the application of the gradient collagen molecules and collagen polypeptide that prepare.
The step 1 of the above method, the pretreatment process method for extracting collagen raw material
In order to ensure the edibility of collagen, extraction method of the present invention based on physical method is to prepare green food
The guarantee of product, by raw material after pretreatment, the collagen of the high-quality of can extraction under certain condition, pretreatment
Method is as follows：It is prepared by the Animal Skin, bone, hyaline cartilage, bone, fish-skin, fish scale or fish-skin (band fish scale) etc. of choosing carefully out first
The material of collagen, respectively with 10%-100% not wait vinegar (can also be wine etc.) soak 0-2 hours, remove part ion and
Amine, during which remove the debris such as hair, fat.Then, the water of immersion is outwelled, weight ratio water 1:0.5-50 adds running water, distillation
Either mineral water etc. is any has or without in treated clear water, the small time-varying of 0.1-10 is boiled under high pressure or non-high-pressure situations for water
Property, or 300-10000g centrifugations remove to combine glycolipid, standby or boil again.
The step 2 of the above method, the technique and technology for extracting collagen macromolecular
Material：Water is 1:0.5-50, boiled under high pressure or non-high-pressure situations, or microwave extracts -10 hours 8 minutes,
Repeat 1-5 times or more time, it is cool to be finally incubated nature, goes to combine glycolipid, renaturation to 30-70 degree, multiple times of filtration acquisition glue
Former albumen.Enzymolysis liquid is carried out refined filtration by this process using filter paper, vertical diatomite filtering machine, will to reach the use of next procedure
Ask.Form the mixing stoste containing the type of collagen I, II type, III type, IV type, V type and Ⅺ type.
The step 3 of the above method, the collagen gradient molecule by collagen macromolecular to 18KDa or so, and
18KDa or so collagen molecules preparation method and operating procedure
By collagen biological enzyme, all protease includes pepsin, trypsase, cathepsin, wood
Melon protease, subtilopeptidase A, neutral proteinase, flavor protease, carboxyl protease and alkali protease etc. all may be used
With by single or be used in mixed way, Partial digestion is formed from collagen macromolecular to 18KDa or so collagen molecules；Entirely
Enzymolysis forms 18KDa or so collagen molecules (such as Fig. 1), and device therefor and auxiliary facility have the pressure-resistant enzymolysis of double-layer heat insulation
Tank, all kinds of glass containers, operating platform, water-bath.
The step 4 of the above method, collagen molecules by 18KDa or so are prepared to the gradient collagen polypeptide for being less than 3KDa
Molecule, and the operating procedure of 3KDa collagen polypeptide molecule
Collagen biological enzyme, increase enzyme amount (enzyme amount for increasing 2-20%) and increase degradation time, device therefor
There are the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers, operating platform, water-bath with auxiliary facility.0.1-10000ml (
Can be more or less) concentration be 0.0001-1000ug/ul collagen, with 0.0005-50mg/ml enzyme, pH5-9 mono-
As be 7, digested after being adjusted in the reaction system of -24 hours 3 minutes (also can longer time or shorter time) enzymolysis, put
Be cooled to room temperature after destroying enzyme activity into 90 DEG C of water-baths, obtain Partial digestion formed from 18KDa or so collagen molecules to
3KDa gradient collagen peptide molecule, and the collagen polypeptide molecule less than 3KDa.
The step 5 of the above method, prepare by collagen polypeptide of the molecular weight less than 3KDa to the relatively small molecule for being less than 1.5KDa
Collagen polypeptide, and the preparation method and operating procedure of collagen oligopeptide less than 1.5KDa, amino acid oligomers and amino acid；
Collagen is placed, device therefor and auxiliary facility at room temperature after 2-20% enzyme amount and long-time are degraded
There are the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers, operating platform, water-bath.0.1-10000ml (also can more or
Less) concentration is 0.0001-1000ug/ul collagen, and with 0.0005-50mg/ml enzyme, pH5-9 is generally 7, carries out 3
Digest after being adjusted in the reaction system of minutes -24 hours (also can longer time or shorter time) enzymolysis, after room temperature is placed, put
It is cooled to room temperature after destroying enzyme activity into 90 DEG C of water-baths, obtains Partial digestion and form collagen molecules by 3KDa or so to small
In 1500Da gradient collagen peptide molecule, and the collagen polypeptide molecule based on collagen polypeptide molecule less than 1500Da, most
After obtain amino acid.Blueness is presented in Fig. 2 SDS protein adhesives is collagen gradient molecule, is shown from macromolecular and each
Molecular grade forms the hangover of blueness, and middle blue zone is exactly the 18KDa molecules that Partial digestion is formed, and the ratio for being below is more uniform
Small peptide gradient molecule hangover, until the small peptide molecule of 1.5KDa collagens, smaller molecule is difficult to show on this electrophoretogram.
The application of the step 6 of the above method, the gradient collagen molecules and collagen polypeptide prepared
(1) the small molecule collagen of this high-quality has obvious cosmetic result to human body skin, is very safe U.S.
Hold good merchantable brand, oral external application is all suitable, and concentration is the 0.01-20% aqueous solution, or is used with other combinations of substances, effect Jiamei.
(2) drink this product 10-200ml, can feel 24 hours it is energetic, without tired sense because, collagen in vivo
Albumen forms and repairing bodily tissue.The growing of body, the renewal of aging tissues, the reparation of injury tissue, additionally it is possible to adjust
Osmotic pressure is saved, the moisture between human normal plasma and tissue fluid constantly exchanges and maintains to balance, and is required for protein as machine
Body is most important " construction material ".
(3) this product is suitable for the effect for resisting various radiation, and experiment proves that 0.01-1% micromolecular collagen stostes just have
The effect of various good anti-various radiation, and good water conservation layer can be formed.
(4) this product can be used as functional health care food prevention of cardiovascular disease, it have improve circulation, to coronary heart disease, lack
Courageous and upright encephalopathic is favourable.There is close relationship with prevention of arterial hardening, hypertension.
(5) this product is assisting body discharge aluminum, and reducing aluminum, aggregation aspect also has unique distinction in vivo.Aluminium is to human body
Harmful, research shows, the senile dementia gradually increased a few days ago is relevant with the intake of aluminium.Contain a large amount of sweet ammonia in collagen
Acid, it is not only involved in rubber polymer original in human body, but also it is a kind of nervous centralis inhibitory transmitter in brain cell to be, to produce
The sedation of Central nervous, there is good therapeutic action collagen food to anxiety disorder, neurasthenia etc..
(6) usually invented food can be with zhuanggu, and the ossein in osteocyte is the binder of hydroxyapatite, and it is imitated with hydroxyl
Apatite together constitutes the main body of bone.Collagen is the important composition composition in skeleton, especially cartilaginous tissue.
Collagen just as one in bone be full of duck eye net, it can firmly keep here will be lost in it is calcareous.Filled without this
The net of full duck eye, even supplemented with excessive calcium, can also be lost in vain.Under normal circumstances, the calcium in sclerotin total amount,
The inorganic substances such as magnesium, phosphorus only account for a few percent of total amount, and the organic substance such as collagen will be accounted for more than 80%.To moving people
For group, the continuous sharp wear in joint can form cartilage damage, cannot recover in a short time or even for good and all.Common people are come
Say, after 25 years old, internal collagen begins to gradually be lost in, especially women, due to hormone in vivo caused by the age
Imbalance, the speed several times faster than male of loss.Therefore the optimum method for maintaining joint and bone health is exactly to supplement in time
Calcareous and collagen.
(7) this product can be as the modifying agent of frozen food, and as jelly, its fusing point is relatively low, is soluble in hot water, has
The characteristics of just melt in the mouth, it is commonly used for meal jelly, grain jelly, jelly etc..Gelatin is general with this product amount in ice cream
For 0.25%-0.60%.
(8) this product can be as the fining agent of beverage, and fish glue is five-star glue fining agent generally acknowledged in the world, in beer and
Grape Wine Industry, fish glue and gelatin can obtain very good effect as sedimentation, fining agent.
(9) this product can be used as stabilizer for dairy products, can make an addition in milk powder, can both improve the nutritive value of milk powder, again may be used
Strengthen the feature of milk powder, accelerate skeleton development, the immunity for the body that increases intelligence, improves.
(10) this product can be used as food additives, and collagen polypeptide may be added to that in candy, bread, addition 3%
~5%, the ageing time of bread can be extended.
Brief description of the drawings
The main product based on 18KDa collagen polypeptide after Fig. 1 SDS protein adhesives macromolecular collagen protein enzymolysis.
Blueness is presented in Fig. 2 SDS protein adhesives is collagen gradient molecule, is shown from macromolecular and each molecule ladder
Degree formed blueness hangover, middle blue zone be exactly Partial digestion formed 18KDa molecules, the more uniform small peptide of ratio for being below
Gradient molecule trails, until to the small peptide molecule of 1.5KDa collagens.
Embodiment：
Embodiment 1, the pretreating process for extracting collagen raw material
In order to ensure the edibility of collagen, extraction method of the present invention based on physical method is to prepare green food
The guarantee of product, by raw material after pretreatment, the collagen of the high-quality of can extraction under certain condition, pretreatment
Method is as follows：It is prepared by the Animal Skin, bone, hyaline cartilage, bone, fish-skin, fish scale or fish-skin (band fish scale) etc. of choosing carefully out first
The material of collagen, respectively with 10%-50% not wait vinegar (can also be wine etc.) soak 0.5-1 hours, remove part ion and
Amine, during which remove the debris such as hair, fat.Then, the water of immersion is outwelled, weight ratio water 1:10-20 adds running water, distilled water
Either mineral water etc. is any has or without in treated clear water, the small time-varying of 0.1-10 is boiled under high pressure or non-high-pressure situations
Property, insulation is until room temperature, and with knife, either other hard things scrape off unnecessary fat etc. or 3000-4000g centrifugations go to combine
Glycolipid, it is standby or boil again, in case the extraction of collagen is used.
Embodiment 2, the technique and technology for extracting collagen in the material:
The Animal Skin, bone, hyaline cartilage, bone, fish-skin, fish scale or fish-skin (band fish scale) etc. of choosing carefully out prepare collagen
Material, carry out collagen extracting, device therefor and auxiliary facility have the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers,
Operating platform, water-bath.
Take：Material：Water is 100:200-500, which adds running water, distilled water either mineral water etc. is any, to be had or nowhere
In the clear water managed, it is molecule degeneration that -3 hours 50 minutes are boiled under high pressure or non-high-pressure situations, is during which incubated at 100 degree,
Until be naturally cooling to room temperature, repeat boil with temperature-fall period 1-5 time or more, finally insulation nature is cool, goes to combine
Glycolipid, renaturation to 40-50 degree, multiple times of filtration obtain collagen.This process will be digested using filter paper, vertical diatomite filtering machine
Liquid carries out refined filtration, to reach the requirement of next procedure.It is filling, as fruit product need to use liquid, the then liquid after filtering
Can direct packaging into standard keg packaging.Storage liquid condition saves backup after the assay was approved, can also concentrate, spray and do
Dry, the liquid after filtering can be just spray-dried after needing concentration to reach finite concentration before entering spray tower, and this is to improving economic effect
The meaning of benefit and production efficiency is very big.
Form the mixing stoste containing the type of collagen I, II type, III type, IV type, V type and Ⅺ type.If necessary to certain type
Number collagen, can be added and subtracted for raw material.
Embodiment 3, the collagen molecules by collagen giant molecule to 18KDa or so, and 18KDa or so glue
Collagen molecules preparation method and operating procedure based on former protein molecular
The collagen biological enzyme that will be obtained in 2, Partial digestion are formed by collagen macromolecular to 18KDa or so
Collagen molecules；The holoenzyme solution under the conditions of simultaneously, forms 18KDa or so collagen molecules, device therefor and matches somebody with somebody
Set facility has the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers, operating platform, water-bath.Specific method and operating procedure are：
Prepared before enzymolysis, according to enzymatic activity specification, configure bromelain or papain etc. enzyme liquid, reference
Prepare 10mL, 1mL packing, stored protected from light (tinfoil paper bag), -20 degree persistences.
Enzymolysis step:Water transfer bath, to digest, in addition, water transfer bath is to 90 DEG C, is used to 55 DEG C to do enzyme inactivation.
Preliminary experiment first is carried out with the collagen and enzyme of small size, adds the enzyme of different volumes, enzymolysis liquid, SDS are taken out in different time
Electrophoresis, observe enzymolysis situation.Afterwards, the dosage of collagen protein enzymolysis volume Ah and's enzyme, the product batch number activity of different enzymes are determined
It is different with dosage.
Such as 0.1-10ml (also can the be more or less) collagen that concentration is 0.01-10ug/ul, 0.05-5mg/ is used
Ml enzyme, pH5-7, after carrying out -2 hours 3 minutes (also can longer time or shorter time) enzymolysis, put into 90 DEG C of water-baths
It is cooled to room temperature after destroying enzyme activity, obtains the collagen molecules by collagen giant molecule to 18KDa or so of Partial digestion；
And the collagen molecules (see Fig. 1) based on 18KDa or so collagen molecules, Fig. 1 SDS protein adhesive macromolecular collagens
The main product based on 18KDa collagen polypeptide after proteolysis.It is cool to be finally incubated nature, multiple times of filtration, is placed on refrigerator storage
Deposit or standby.Enzymolysis liquid is carried out refined filtration by this process using filter paper, vertical diatomite filtering machine, to reach making for next procedure
With requiring.It is filling, as fruit product need to use it is liquid, then filter after liquid can direct packaging into standard keg packaging.Inspection
Test warehousing after passing liquid condition to save backup, can also concentrate, be spray-dried, the liquid after filtering needs dense before spray tower is entered
Contracting can be just spray-dried after reaching finite concentration, and this is to increasing economic efficiency and the meaning of production efficiency is very big.
Embodiment 4, prepare by 18KDa or so collagen molecules to be less than 3000Da gradient collagen peptide molecule,
And the method and operating procedure of the collagen polypeptide molecule based on the collagen polypeptide molecule less than 3KDa
The collagen biological enzyme that will be obtained in 2, when increasing enzyme amount (enzyme amount for increasing 2-10%) and increasing degraded
Between, device therefor and auxiliary facility have the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers, operating platform, water-bath.In 0.1-
The collagen that 10ml (also can be more or less) concentration is 0.01-10ug/ul, with 0.05-50mg/ml enzyme, pH5-9 mono-
As be 7, digested after being adjusted in the reaction system of 1-2 hours (also can longer time or shorter time) enzymolysis, put to 90
It is cooled to room temperature after enzyme activity is destroyed in DEG C water-bath, obtains Partial digestion and form collagen molecules by 18KDa or so to being less than
3KDa gradient collagen peptide molecule, and the collagen polypeptide molecule based on collagen polypeptide molecule less than 3KDa, II Collagen Type VI
Collagen macromolecular has been obtained after the thermal degradation reaction of albumen to the collagen polypeptide of the dalton of molecular weight 2450, has been prepared
The equal relative molecular weight of product number is less than 3000 dalton.Multiple times of filtration, it is placed on refrigerator storage or standby.This process utilizes filter
Enzymolysis liquid is carried out refined filtration by paper, vertical diatomite filtering machine, to reach the requirement of next procedure.It is filling, as fruit product needs
Using liquid, then liquid after filtering can direct packaging into standard keg packaging.Liquid condition is put in storage after the assay was approved to protect
Deposit standby, can also concentrate, be spray-dried, the liquid after filtering just can be with after needing concentration to reach finite concentration before entering spray tower
Spray drying, this is to increasing economic efficiency and the meaning of production efficiency is very big.
Embodiment 5, prepare by molecular weight less than 3KDa collagen polypeptide to be less than 1.5KDa it is more compared with the collagen of small molecule
Peptide, and the preparation method and operating procedure of collagen oligopeptide less than 1.5KDa, amino acid oligomers and amino acid
Collagen can be by specific proteasome degradation, i.e. biological degradability.Because collagen has the helical structure of close and firm, institute
Its side chain can only be cut off with most protease, the only specific protease such as clostridiopetidase A, elastoser is in specific bar
Could degrade collagen under part, be broken collagen peptide bond.For the peptide bond of collagen once being broken, its helical structure is destroyed and thorough immediately
Bottom is hydrolyzed to micromolecule polypeptide or amino acid, so as to obtain gradient collagen and the small peptide molecule from macromolecular to 1500 or so
(see Fig. 2), blueness is presented in Fig. 2 SDS protein adhesives is collagen gradient molecule, is shown from macromolecular and each molecule ladder
Degree formed blueness hangover, middle blue zone be exactly Partial digestion formed 18KDa molecules, the more uniform small peptide of ratio for being below
Gradient molecule trails, and is difficult to show on this electrophoretogram until to the small peptide molecule of 1.5KDa collagens, smaller molecule.
The collagen biological enzyme that will be obtained in 4, when increasing enzyme amount (enzyme amount for increasing 5-20%) and increasing degraded
Between, device therefor and auxiliary facility have the pressure-resistant enzymatic vessel of double-layer heat insulation, all kinds of glass containers, operating platform, water-bath.In 0.1-
The collagen that 10000ml (also can be more or less) concentration is 0.0001-1000ug/ul, with 0.0005-50mg/ml's
Enzyme, pH5-7.5, carry out enzyme after being adjusted in the reaction system of -24 hours 3 minutes (also can longer time or shorter time) enzymolysis
Solution, put into 90 DEG C of water-baths and be cooled to room temperature after destroying enzyme activity, obtain Partial digestion and formed and divided by 3KDa or so collagen
Son arrives the gradient collagen peptide molecule less than 1500Da, and the collagen polypeptide point based on collagen polypeptide molecule less than 1500Da
Son, finally to the preparation method of amino acid, see in 4, collagen macromolecular adds to the collagen polypeptide of the dalton of molecular weight 2450
Big enzyme amount and reaction time, the equal relative molecular weight of product number being prepared is smaller, or even obtains single amino acids.This process profit
Enzymolysis liquid is subjected to refined filtration with filter paper, vertical diatomite filtering machine, to reach the requirement of next procedure.It is filling, if production
Product need to use it is liquid, then filter after liquid can direct packaging into standard keg packaging.Be put in storage after the assay was approved liquid
State saves backup, and can also concentrate, be spray-dried, the ability after needing concentration to reach finite concentration before entering spray tower of the liquid after filtering
It can be spray-dried, this is to increasing economic efficiency and the meaning of production efficiency is very big.It is placed on refrigerator storage or standby.
Application of the embodiment 6 in beauty industry
This product is i.e. absorbable in 30 seconds after painting face, can play prevention collagenous fibres connection solidification, increase iuntercellular
Mucopolysaccharide, anti-aging, wrinkle resistant, the effect of nti-freckle.
With advancing age, fibroblastic synthesis capability declines, if lacking collagen in skin, collagenous fibres
Connection solidification will occur, reduce iuntercellular mucopolysaccharide, skin will lose soft, elasticity and gloss, and aging, collagen egg occurs
White is Animal Skin extraction mostly, contains the materials such as hyaluronic acid, chondroitin sulfate in skin, and they contain a large amount of polar groups,
It is moisturizing factor, and plays the role of to prevent the tyrosine in skin from being converted into melanin, while prevents the fibrous fracture of corium, fat
Fat atrophy, sweat gland and smegma are reduced, and skin a series of aging phenomena such as color spot, wrinkle is occurred.Therefore collagen has
Pure natural moisturizing, whitening, wrinkle resistant, nti-freckle etc. act on, and collagen is similar to the structure of human body skin collagen, rich in big in molecule
Amino acid and hydrophilic group are measured, there is certain surface-active and good compatibility, simultaneously because containing substantial amounts of in its molecule
Hydroxyl, therefore it has fairly good nutrition and moisture-keeping function.Drink and smear the depth that can be spread to tissue, human body, skin
Layer, the integrality of cuticula moisture and fibre structure is kept, promotes the metabolism of skin histology, skin is produced well
Moisten moisturizing, effect wrinkles eliminating vanishing effect.Early in the early 1970s, the U.S. just takes the lead in releasing injection bovine collagen, for nti-freckle
Wrinkle and reparation scar.
This product gradient collagen, there is obvious cosmetic result to human body skin, is very safe cosmetic good merchantable brand, mouth
Take that external application is all suitable, concentration is the 0.01-20% aqueous solution, or is used with other combinations of substances, effect Jiamei.
Various toners (containing protocollagen 0.5-3%), cream (containing protocollagen 0.1-3%), frost is made
All kinds of skin care item of (containing protocollagen 0.5-3%).
Embodiment 7, the application in terms of packaging material for food
Inner packing films such as the casing products of all kinds of sausage goods, candied fruit etc. can be prepared with collagen, collagen casing
Itself it is nutritious high protein material.In heat treatment process, as the evaporation of moisture and grease is with dissolving.Collagen is almost
It is consistent with the shrinkage factor of meat product, and other edible package materials are not found to have this quality also.Collagen
Artificial casing also has the characteristics that in good taste, transparency is high, manufacture craft is simple simultaneously, good beneficial to a large amount of productions, DEVELOPMENT PROSPECT.
In 0.5-2% collagens, it is heated to dissolving, the solution of calcium chloride is added dropwise entered afterwards, form cross-linking reaction,
It is paved into film and forms layer of gel film, for packaging for foodstuff.As not only attractive in appearance but also have good mechanical performance, while it is also a kind of battalion
Support carrier, the collagen packaging film of edibility, typically with good tensile strength, heat sealability and higher choke, oil resistance,
Hinder moist etc., can be widely applied to the fresh-keeping of varieties of food items.
Embodiment 8, collagen protein health protection food
This product takes high temperature water-boiling method to extract collagen, can directly eat, and can strengthen immunity function, and increase is immune
Power.
Collagen improves body immunity and reaches compacting cancer cell purpose, is also prevented from cancer metastasis.By this hair
Bright method is prepared by collagen, aids in preparing nourishing liquid with white fungus extract, 100ml prepares matching somebody with somebody for nourishing liquid by collagen
Side：
This product collagen 30ml, with mineral water, distilled water or clean water：70ml (is adjusted) according to taste and suitable sensitivity.
Sugar or sweetener：Carbohydrate, honey either xylitol (according to whether being diabetes or selection)
Food additives：Sodium benzoate in the range of national standard and, or sorbic acid and sodium sorbate, flavoring essence.
Any other active principle, such as a series of materials of matrimony vine, jujube can be also added, is prepared into oral liquid.
This product can show that collagen can reduce blood glycerine as functional health care food with prevention of cardiovascular disease, research
Three esters and cholesterol, while collagen has the effect of accelerating hemoglobin and RBC acceptor garland rate, it, which has to improve, circulates, to hat
Worry, ischemic cerebral disease are favourable.There is close relationship with prevention of arterial hardening, hypertension.
Embodiment 9, the production technology for the beverage that the person of collectively forming in osteocyte recovers with bone, joint：
Collagen is the important composition composition in skeleton, especially cartilaginous tissue.Collagen is just as in bone
One be full of duck eye net, it can firmly keep here will be lost in it is calcareous.The net of duck eye is full of without this, is even mended
The calcium of excess has been filled, can be also lost in vain.Under normal circumstances, the inorganic substances such as the calcium in sclerotin total amount, magnesium, phosphorus only account for always
A few percent of amount, and the organic substance such as collagen will be accounted for more than 80%.For sport people, joint it is continuous acutely
Abrasion can form cartilage damage, cannot recover in a short time or even for good and all.Replenishing collagen can improve joint, cartilage and
The lubrication of ligament, mitigate anchylosis, ache, the symptom such as ponding.
Embodiment 10, one kind are prepared the production technology of beverage by collagen, white fungus " food materials elite " extract：
This technique strict implement sterileization product specification.
Production technology is：
B liquid is extracted, 1-50 parts, temperature is 0-100 degree, adds 0-10 part A liquid, and temperature is unlimited, adds food additives：State
Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of mark.Any other active principle can also be added, as matrimony vine,
A series of materials of jujube etc. or extract.80-99.9 parts, 4-100 degree distilled water or mineral water or pure water are added, it is fully mixed
Close, sterilizing, be distributed into bottle.
This production technology can require according to different production lines, be improved, still, it is desirable to it is final sterile, or strict symbol
Close country, international standard.
A in taking above-mentioned 1,2, B liquid, it is a kind of by the preparation of collagen, white fungus extract and mixture to prepare " food materials elite "
The formula of beverage.
(exemplified by 100 milliliters, can expand and reduce) is：
Extract A liquid：30ml (this product collagen)
Extract B liquid：20ml (white fungus extract)
(single plus a kind of also can)
Mineral water, distilled water or clean water：50ml (is adjusted) according to taste and suitable sensitivity
Sugar or sweetener：Carbohydrate, honey either xylitol (according to whether being diabetes or selection)
Food additives：Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of 0- national standards.
Any other active principle, such as a series of materials of matrimony vine, jujube can also be added.
The beverage and nourishing liquid prepared by collagen, white fungus is formed, can be in the air pollutions such as smoker and haze
People has effect of clearing away lung-heat.
Embodiment 11, prevent cardiovascular and cerebrovascular disease and the beverage and oral tonic liquor of part malignant tumour：
The gradient collagen of the present invention is a kind of preferable blood fat reducing food, can reduce blood triglyceride and courage is consolidated
Alcohol, and the essential trace element that can increase internal some shortages is allowed to maintain within a relative normal range (NR).II type
Collagen is the major protein in articular cartilage, thus is potential autoantigen.Energy inducing T cell produces immune resistance to after oral
By so as to suppress the autoimmune disease of T cell mediation.
White fungus has the effect of cardiac stimulant strengthens body and anticancer.It can not only improve liver detoxification, moreover it is possible to it is anti-swollen to strengthen body
The immunocompetence of knurl, and tumor patient can be strengthened to radiotherapy, the tolerance of chemotherapy.White fungus is kind of the fat-reducing food containing dietary fiber
Product, its dietary fiber can help gastrointestinal peristalsis, reduce fat absorption.
100ml (exemplified by 100 milliliters, can expand and reduce) prevents cardiovascular and cerebrovascular disease and the beverage of part malignant tumour
And oral tonic liquor：
Extract A liquid：25ml (this product collagen)
Extract B liquid：15ml (other Chinese medicine class additives)
(single plus a kind of also can)
Mineral water, distilled water or clean water：60ml (is adjusted) according to taste and suitable sensitivity
Sugar or sweetener：Carbohydrate, honey either xylitol (according to whether being diabetes or selection)
Food additives：Sodium benzoate in the range of 0- national standards and, or sorbic acid and sodium sorbate, flavoring essence.
Any other active principle, such as a series of materials of matrimony vine, jujube can also be added.
The beverage and nourishing liquid prepared by collagen, white fungus etc. " food materials elite " is formed, cardiovascular and cerebrovascular disease can be prevented
With the beverage and oral tonic liquor of part malignant tumour.
Embodiment 12, reduce the beverage and oral tonic liquor of the aggregation of aluminium in vivo：
There are some researches show collagen can assist to exclude internal aluminium, reduce the aggregation of aluminium in vivo, reduce aluminum pair
The harm of human body.
White fungus has promoting production of body fluid and nourishing the lung, tonifying Qi and blood, strong mesh kidney tonifying, clear and rich beneficial stomach, brain refreshing, promotes longevity, cardiac stimulant is strengthened
The effect of body and anticancer.
The formula (100 milliliters) that collagen, white fungus extract and mixture prepare beverage is：
Extract A liquid：15ml (this product collagen)
Extract B liquid：15ml (other nutriments and sweetener etc.)
(single plus a kind of also can)
Mineral water, distilled water or clean water：70ml (is adjusted) according to taste and suitable sensitivity
Sugar or sweetener：Xylitol
Food additives：Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of national standard.
Any other active principle, such as a series of materials of matrimony vine, jujube can also be added.
B liquid is extracted, 1-60 parts, temperature is 0-100 degree, adds 0-20 part A liquid, and temperature is unlimited, adds food additives：0-
Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of national standard.Any other active principle, such as Chinese holly can also be added
A series of materials of Qi, jujube etc. or extract.70-99.0 parts, 4-100 degree distilled water or mineral water or pure water are added, fully
Mixing, that is, form food materials elite --- and moisturizing is wrinkle resistant, pre- anti-aging beverage and oral tonic liquor.
Embodiment 13, prevention arthralgia, the beverage and oral tonic liquor for improving immunity：
Collagen also can be used for the beverage and oral tonic liquor of prevention arthralgia, early in 12nd century Bingen
St.Hilde-gard is just described by the use of the cartilage soup of calf as medicine to treat arthralgia, in considerably long one section
Between in, some products containing collagen are by it is believed that be very helpful to joint.Collagen is skeleton, especially soft
Important composition composition in bone tissue.Collagen is full of the net of duck eye just as one in bone, and it can firmly keep here just
What is be lost in is calcareous.The net of duck eye is full of without this, even supplemented with excessive calcium, can be also lost in vain.
White fungus has promoting production of body fluid and nourishing the lung, tonifying Qi and blood, strong mesh kidney tonifying, clear and rich beneficial stomach, brain refreshing, promotes longevity, cardiac stimulant is strengthened
The effect of body and anticancer.It can not only improve liver detoxification, moreover it is possible to strengthen the antitumor immunocompetence of body, and can strengthen
Tumor patient is to radiotherapy, the tolerance of chemotherapy.White fungus is kind of the diet food containing dietary fiber, and its dietary fiber can help stomach and intestine
Wriggle, reduce fat absorption.Both food materials are suitable for all ages, form few person in middle and old age and often use beverage：
It can by a certain percentage increase and decrease, or add other materials:
B liquid is extracted, 1-50 parts, temperature is 0-100 degree, adds 0-10 part A liquid, and temperature is unlimited, adds food additives：0-
Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of national standard.Any other active principle, such as Chinese holly can also be added
A series of materials of Qi, jujube etc. or extract.80-99.9 parts, 4-100 degree distilled water or mineral water or pure water are added, fully
Mixing, sterilizing, is distributed into bottle.
Embodiment 14, impaired or allergy epidermis reparation person often use beverage and nourishing liquid：
Collagen is extracted by Animal Skin, also containing albumen such as hyaluronic acid, chondroitin sulfates in addition to collagen in skin
Polysaccharide, they contain a large amount of polar groups, are moisturizing factors, and have the work for preventing the tyrosine in skin from being converted into melanin
With, therefore collagen has the effect such as pure natural moisturizing, whitening, wrinkle resistant, nti-freckle, can be widely applied in cosmetic products.Collagen egg
White chemical composition, structure impart it be beauty basis.Collagen is similar to the structure of human body skin collagen, is non-aqueous
Soluble fiber shape contains glycoprotein, and a large amount of amino acid and hydrophilic group are rich in molecule, with certain surface-active and well
Compatibility, simultaneously because contain substantial amounts of hydroxyl in its molecule, therefore it has fairly good moisture-keeping function.In relative humidity
When 70%, the moisture of its own weight 45% can be still kept.Experiment proves：The 0.01% pure solution of collagen can just be formed very
Good water conservation layer, supply whole moisture required for skin.
With advancing age, fibroblastic synthesis capability declines, if lacking collagen in skin, collagenous fibres
Connection solidification will occur, reduce iuntercellular mucopolysaccharide, skin will lose soft, elasticity and gloss, and aging occurs, while very
Fibrous fracture, lipoatrophia, sweat gland and the smegma of skin are reduced, and skin a series of agings such as color spot, wrinkle is occurred and are showed
As.When being used for as active material in cosmetics, the latter can be spread to the deep layer of skin, its tyrosine and skin that contain
Tyrosine competition in skin, and combined with the catalytic center of tyrosinase, so as to suppress the generation of melanin, make the glue in skin
Former protein active enhancing, keeps the integrality of cuticula moisture and fibre structure, promotes the metabolism of skin histology, to skin
Skin produces good moist moisturizing, effect wrinkles eliminating vanishing effect.Early in the early 1970s, the U.S. just takes the lead in releasing injection ox glue
Original, for nti-freckle wrinkle and repair scar.
It is simple to usually require that molecular weight in below 2KD as trophism skin type raw material but in cosmetics, to allow water
Dispergation proper energy is infiltrated through in skin.And hair care category cosmetics should also have certain in addition to requiring that hydrolytic collagen has moisture retention
Film forming, therefore, the molecular weight requirement of hydrolytic collagen can be higher.
Tremella polysaccharides is considered a kind of plant-derived hyaluronic acid-like material, and molecular weight reaches 100-130 ten thousand, is
So far, unique molecular weight can reach the NMF of more than 1,000,000 plant origin in cosmetic material circle.With it is saturating
When bright matter acid is compared, the effect of many surmounts hyaluronic acid has been embodied.White fungus has (1) moisture retention：Its moisturizing
Action principle is to form hydrogen bond by numerous terminal hydroxyl and water molecules, so as to combine substantial amounts of water, in space shape
Into a spongiform structure of molecule.Such moisturizing principle has also been bonded popular one of current state-of-the art water imbibition NMF
Individual trend.(2) it is anti-oxidant：, tremella polysaccharides is in addition to fabulous water retention, additionally it is possible to and the activity of inhibitory activity oxygen,
Play oxidation resistant effect.(3) film forming：The aqueous solution of tremella polysaccharides and Sodium Hyaluronate is placed in evaporating dish and done
Dry, the film shrinkage factor of tremella polysaccharides is only 10%, and the film shrinkage factor of Sodium Hyaluronate has nearly reached half, it was demonstrated that white fungus is more
Sugared film does not shrink with the change of humidity, so not having tight sensation.Tremella polysaccharides has good affinity to skin,
Make skin become moistening, it is smooth.
100ml " food materials elite " is impaired or the reparation person of allergy epidermis is often as follows with beverage and nourishing liquid and preparation method thereof：
Extract A liquid：30ml (protocollagen product)
Extract B liquid：15ml (white fungus Aqueous extracts)
(single plus a kind of also can)
Mineral water, distilled water or clean water：30-90ml (is adjusted) according to taste and suitable sensitivity
Sugar or sweetener：Carbohydrate, honey either xylitol (according to whether being diabetes or selection)
Food additives：Sodium benzoate in the range of national standard and, or sorbic acid and sodium sorbate, flavoring essence.
Any other active principle, such as a series of materials of matrimony vine, jujube can also be added.
Embodiment 15, application in the beverage
Gradient collagen has degradability, stability and absorbability.
Collagen can be by specific proteasome degradation, i.e. biological degradability.Because collagen has the helical structure of close and firm, institute
Its side chain can only be cut off with most protease, the only specific protease such as clostridiopetidase A, elastoser is in specific bar
Could degrade collagen under part, be broken collagen peptide bond.For the peptide bond of collagen once being broken, its helical structure is destroyed and thorough immediately
Bottom is hydrolyzed to micromolecule polypeptide or amino acid, and small-molecule substance can enter blood circulation system, by body re-using or generation
Thank to discharge.Collagen is human body nature albumen, has larger affinity, weaker anti-to the protein molecule of skin surface
Originality, good biocompatibility and biodegradable security, degradable absorption, adhesion strength are good.Available for beauty, food.
White fungus is sweet, light, mild-natured, nontoxic, existing nourishing Yin and moistening lung, the effect of QI invigorating gut purge, and can enhancing body immunity,
Strengthen tolerance of the tumor patient to Radiotherapy chemotherapy.White fungus has the function that enriching yin, is rich in natural plant colloid, additional its
Good moisturizing food that can be long-term use of.Skin elasticity, plentiful hypodermis can be strengthened, reduce smooth out wrinkles, make skin
Seem delicate smooth, and effectively prevent the evaporation of moisture and virus from invading and making from skin pore with toxic dust in air
The harm of paired human body, the tremella polysaccharides that molecular weight reaches 100-130 ten thousand is a kind of food materials source property hyaluronic acid-like material.
The formula (100 milliliters) that collagen, white fungus extract and mixture prepare beverage is：
Extract A liquid：40ml (protocollagen product)
Extract B liquid：10ml (other sugar or sweetener regulation liquid material)
(single plus a kind of also can)
Mineral water, distilled water or clean water：50ml (is adjusted) according to taste and suitable sensitivity
Sugar or sweetener：Carbohydrate, honey either xylitol (according to whether being diabetes or selection)
Food additives：Citric acid, sodium citrate, sodium benzoate and flavoring essence in the range of national standard.
Any other active principle, such as a series of materials of matrimony vine, jujube can also be added.
Embodiment 16, food additives
Collagen is rich in 18 kinds of amino acid in addition to tryptophan and cysteine, wherein necessary to maintaining growth in humans
Amino acid has 7 kinds, is deposited in other protein also containing hydroxyproline and pyroglutamic acid rare in general albumen and hardly
Oxylysine, nutrition very abundant.
Collagen mouthfeel is soft, and as food additives, taste is light, easy to digest.Take the gradient of appropriate (1%-3%)
Collagen, which has, significantly enhances granulating effect, is modulated into different tastes, for soup, the additive of beverage, there is strongly hydrophilic
Can, it is abundant proteinaceous nutrient source, can also be with beauty and health care and other effects.
Claims (9)
- A kind of 1. preparation method of gradient collagen and collagen polypeptide molecule, it is characterised in that：Comprise the following steps：(1) pretreating process of collagen raw material is extractedFirst by raw material vinegar or soaking in Chinese liquor, remove part ion and amine, during which remove hair, fat debris；Then, will soak Liquid outwell, add water, the small time variations of 0.1-10 are boiled under high pressure or non-high-pressure situations；(2) technique for extracting collagen macromolecularMaterial:The mass ratio of water is 1:0.5-50, collagen extract -10 hours 8 minutes, repeat 1-5 times, be finally incubated Naturally it is cool, go to combine glycolipid, renaturation to 30-70 degree, multiple times of filtration obtains collagen；Formed containing the type of collagen I, II Type, III type, IV type, the mixing stoste of V type and Ⅺ type；(3) by the collagen gradient molecule of collagen macromolecular to 18KDa or so, and 18KDa or so collagen Molecule preparation method and operating procedureBy the collagen biological enzyme of step (2) acquisition, Partial digestion formation is from collagen macromolecular to 18KDa or so Collagen gradient molecule；All enzymolysis forms 18KDa or so collagen molecules；(4) collagen molecules by 18KDa or so are prepared to the gradient collagen peptide molecule for being less than 3KDa, and 3KDa glue The operating procedure of former peptide moleculeThe collagen biological enzyme that will be obtained in step (2), it is 0.0001-1000ug/ul's in 0.1-10000ml concentration In collagen, with 0.0005-50mg/ml enzyme, pH5-9, carry out after being adjusted in the reaction system of -24 hours 3 minutes enzymolysis Enzymolysis, put into 90 DEG C of water-baths and be cooled to room temperature after destroying enzyme activity, obtain collagen of the Partial digestion formation from 18KDa or so Molecule is to 3KDa gradient collagen peptide molecule, and the collagen polypeptide molecule less than 3KDa；(5) prepare and 3KDa collagen polypeptide is less than to the collagen polypeptide of the relatively small molecular weight less than 1.5KDa by molecular weight, and The preparation method and operating procedure of collagen oligopeptide, amino acid oligomers and amino acid less than 1.5KDaIn the collagen that 0.1-10000ml concentration is 0.0001-1000ug/ul, with 0.0005-50mg/ml enzyme, pH5- 9, digested after being adjusted in the reaction system of -24 hours 3 minutes enzymolysis, after room temperature is placed, put into 90 DEG C of water-baths and destroy It is cooled to room temperature after enzyme activity, obtains Partial digestion and form collagen molecules by 3KDa or so to the gradient collagen for being less than 1500Da Peptide molecule, and the collagen polypeptide molecule based on collagen polypeptide molecule less than 1500Da, finally obtain amino acid oligomers And amino acid.
- 2. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： Raw material is Animal Skin, bone, hyaline cartilage, bone, fish-skin, fish scale or fish-skin in the step (1).
- 3. the pretreatment side of the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule Method, it is characterised in that：The weight of vinegar or white wine is the 10%-100% of raw material in the step (1), and soak time is that 0-2 is small When.
- 4. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： It is 0.5-50 that the amount of water and the weight ratio of raw material are added in the step (1):1.
- 5. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： 300-10000 is may also include after being boiled in the step (1) leave the heart and go to combine glycolipid.
- 6. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： The extracting of collagen can use Extract by boiling under high pressure or non-high-pressure situations, or microwave extracting in the step (2).
- 7. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： Filter type uses filter paper or vertical diatomite filtering in the step (2).
- 8. the preparation method of a kind of gradient collagen according to claim 1 and collagen polypeptide molecule, it is characterised in that： Protease in the step (3), (4), (5) used in biological enzyme is pepsin, trypsase, cathepsin, pawpaw It is any one in protease, subtilopeptidase A, neutral proteinase, flavor protease, carboxyl protease and alkali protease Kind, or a variety of enzymes are used in mixed way.
- 9. a kind of application of gradient collagen and collagen polypeptide molecule that preparation method obtains as claimed in claim 1, it is special Sign is：(1) application in beauty industry；(2) prevent collagenous fibres connection solidification, increase iuntercellular mucopolysaccharide, anti-aging, prevent Wrinkle, nti-freckle drink；(3) application in terms of packaging material for food；(4) application of collagen protein health protection food；(5) heart and brain are prevented The beverage and oral tonic liquor of vascular diseases and part malignant tumour；(6) beverage and nourishing mouth of the aggregation of aluminium in vivo are reduced Take liquid；(7) prevent arthralgia, improve the beverage and oral tonic liquor of immunity；(8) impaired or allergy epidermis reparation person Conventional beverage and nourishing liquid；(9) application in the beverage；(10) application in food additives.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109430650A (en) * | 2018-11-22 | 2019-03-08 | 天津市天大天福生物技术有限公司 | Prepare the formula and production technology of collagen, hyaluronic acid beverage and nourishing liquid |
| CN109457006A (en) * | 2018-11-26 | 2019-03-12 | 天津市天大天福生物技术有限公司 | A kind of convenient preparation method of the gradient peptide of different molecular weight mixing |
| CN109627325A (en) * | 2019-01-04 | 2019-04-16 | 广东海洋大学 | A kind of preparation method and applications of gradient collagen and collagen polypeptide molecule |
| CN111995671A (en) * | 2020-09-07 | 2020-11-27 | 青海瑞肽生物科技有限公司 | Collagen peptide and application thereof |
| WO2020245809A3 (en) * | 2019-06-07 | 2021-01-14 | Vivatis Pharma Gmbh | Identification and selection of a plant starting material of a plant chondroitin sulfate and hyaluronic acid, and transformation of such plant starting material to obtain ingredients for use in foods, supplements, medical devices or drugs |
| CN114317661A (en) * | 2022-02-15 | 2022-04-12 | 无锡贝迪生物工程股份有限公司 | Method for preparing multiple active collagen |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109430650A (en) * | 2018-11-22 | 2019-03-08 | 天津市天大天福生物技术有限公司 | Prepare the formula and production technology of collagen, hyaluronic acid beverage and nourishing liquid |
| CN109457006A (en) * | 2018-11-26 | 2019-03-12 | 天津市天大天福生物技术有限公司 | A kind of convenient preparation method of the gradient peptide of different molecular weight mixing |
| CN109627325A (en) * | 2019-01-04 | 2019-04-16 | 广东海洋大学 | A kind of preparation method and applications of gradient collagen and collagen polypeptide molecule |
| WO2020245809A3 (en) * | 2019-06-07 | 2021-01-14 | Vivatis Pharma Gmbh | Identification and selection of a plant starting material of a plant chondroitin sulfate and hyaluronic acid, and transformation of such plant starting material to obtain ingredients for use in foods, supplements, medical devices or drugs |
| CN114174349A (en) * | 2019-06-07 | 2022-03-11 | 维瓦蒂斯制药公司 | Identification and selection of plant starting materials of the plant chondroitin sulphate and hyaluronic acid, and transformation of the plant starting materials to obtain ingredients for food, supplements, medical devices or drugs |
| CN111995671A (en) * | 2020-09-07 | 2020-11-27 | 青海瑞肽生物科技有限公司 | Collagen peptide and application thereof |
| CN111995671B (en) * | 2020-09-07 | 2023-06-20 | 青海瑞肽生物科技有限公司 | Collagen peptide and application thereof |
| CN114317661A (en) * | 2022-02-15 | 2022-04-12 | 无锡贝迪生物工程股份有限公司 | Method for preparing multiple active collagen |
| CN114317661B (en) * | 2022-02-15 | 2024-04-09 | 无锡贝迪生物工程股份有限公司 | Method for preparing multiple active collagen |
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Application publication date: 20171229 |