CN109627323A - Snakeskin collagen peptide and its preparation and application - Google Patents
Snakeskin collagen peptide and its preparation and application Download PDFInfo
- Publication number
- CN109627323A CN109627323A CN201811614637.4A CN201811614637A CN109627323A CN 109627323 A CN109627323 A CN 109627323A CN 201811614637 A CN201811614637 A CN 201811614637A CN 109627323 A CN109627323 A CN 109627323A
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- snakeskin
- collagen
- parts
- collagen peptide
- peptide
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Abstract
本发明涉及蛋白活性肽技术领域，具体涉及蛇皮胶原蛋白肽及其制备和应用，本发明使用经提纯的蛇皮胶原蛋白为酶切原料，经单酶切、多酶复合酶切得到分子量为100Da‑20KDa蛇皮胶原蛋白肽，其具有良好的抗氧化活性，1mg/ml浓度的蛇皮胶原蛋白肽DPPH清除率达到49.36％，超氧阴离子清除率达到51.12％，本发明提供的蛇皮胶原蛋白肽制备方法酶解过程工艺简单，酶切效率高，生产周期短，蛇皮肽得率在52％以上，抗氧化活性强，应用本发明于化妆品领域制得一种抗皮肤老化化妆品，具有显著的抗氧化活性。The invention relates to the technical field of protein active peptides, in particular to snakeskin collagen peptides and their preparation and application. The invention uses purified snakeskin collagen as a raw material for enzymatic cleavage, and single-enzyme cleavage and multi-enzyme compound cleavage obtain a molecular weight of The 100Da-20KDa snakeskin collagen peptide has good antioxidant activity, the DPPH clearance rate of the snakeskin collagen peptide at a concentration of 1 mg/ml reaches 49.36%, and the superoxide anion clearance rate reaches 51.12%. The snakeskin collagen provided by the present invention The protein peptide preparation method has the advantages of simple enzymatic hydrolysis process, high enzymatic cleavage efficiency, short production cycle, more than 52% yield of snake skin peptide, and strong antioxidant activity. The invention is applied to the cosmetic field to prepare an anti-aging cosmetic product, which has the advantages of Significant antioxidant activity.
Description
Technical field
The present invention relates to protein active peptide technical fields, and in particular to a kind of snakeskin collagen peptide and its preparation and answers
With, using the anti-aging cosmetics of snakeskin collagen peptide.
Background technique
Peptide is that one kind is related to the bioactive substance of various kinds of cell function in organism, participates in the various complexity of regulation human body
Physiology course, it is active high, compatibility is strong and highly-safe the features such as, and the peptide molecule of micron level, it is easy to pass through skin
Peptide surface layer is directly entered deep layer skin peptide, is quickly absorbed by the body and plays a role.Modern pharmacology research shows some active peptides not
Only there is antioxidant activity, the growth and proliferation of human skin fibroblasts can also be effectively facilitated, can especially pass through increasing
The activity for adding skin fibroblasts effectively facilitates the synthesis of the matrix such as cellular matrix especially collagen, plays significant
Resisting age of skin effect.
Numerous studies discovery, the antioxidation of peptide is mainly some small molecules being made of 2-10 amino acid residue
Peptide, antioxidant activity are mainly determined by following factor: (1) special acid residue, such as leucine, histidine, color ammonia
Acid, methionine and cysteine etc., they are directly related with antioxidant activity；(2) amino acid sequence directly determines active peptide
Antioxidant activity, amino acid sequence is different, and antioxidant peptide active is also different；(3) hydrophobicity of anti-oxidation peptide or in the solution
Dispersibility；(4) stability of anti-oxidation peptide；(5) barrier function, since peptide is a kind of amphiphilic surfactant, they can shape
Water and oil are separated at the film of a thickness, to prevent fat from directly contacting with free radical and other oxides.In conclusion
Biologically active peptide provides completely new direction and thinking for the development of cosmetics for resisting age of skin and health products trade, in anti-skin
Application in ageing prod is also more and more common.
For collagen as extracellular matrix protein, immunogenicity is low, and property is stablized, cannot be by most protease
Hydrolysis, collagen is similar to the structure of human skin collagen, and amino acid side chain contains a large amount of hydrophilic groups, can be well soluble in
Water, and moisture retention good with skin-friendliness are good.Therefore, also there is better anti-oxidant work using active peptide prepared by collagen
Property and biocompatibility.
Snakeskin is common Chinese medicine, has skin care effect, and snakeskin collagen, which can become, prepares active peptide
Fine raw material, Chinese patent literature CN102277405A disclose a kind of preparation method of snakeskin active peptide, are original with fresh snakeskin
Material, cleaned, rubbing, adjustment material concentration, adjustment temperature of charge, are added protease hydrolyzed, centrifugal filtration, enzyme deactivation, receive defibrination
The process flows such as desalination and concentration, Ultrafiltration Purifying and freeze-drying are filtered, snakeskin active peptide is obtained, 90% or more divides in molecular weight product
It is distributed between 200-50000Da, also containing ingredients such as DHA, EPA.Although this method uses protease hydrolyzed snakeskin collagen,
Simple process, with short production cycle, active peptide obtained has excellent water solubility, stability and moisture-absorbing moisture-keeping function, but its enzyme
It is low to cut resulting snakeskin active peptide yield, effective small-molecular peptides type is few, and antioxidant activity is not high, and obtained product contains
There are more invalid components, separation process is many and diverse, and specific aim is not strong.
Summary of the invention
Therefore, the technical problem to be solved in the present invention is that overcome in the prior art snakeskin collagen peptide by fresh snakeskin group
It is woven to raw material to be digested, causes snakeskin active peptide effective active peptide yield obtained low, the not high technology of antioxidant activity lacks
Fall into, thus provide it is a kind of use snakeskin collagen for raw material, with high purity, the snakeskin collagen peptide yield height of acquisition, antioxygen
Change the good snakeskin collagen peptide and preparation method thereof of performance.
For this purpose, the snakeskin collagen peptide uses snakeskin collagen egg the invention discloses a kind of snakeskin collagen peptide
It is white to be prepared, molecular weight 100Da-20KDa.
Preferably, the molecular weight of the snakeskin collagen peptide is 3KDa-10KDa；
Preferably, the snakeskin is selected from zaocys dhumnade, pallas pit viper, poisonless snake, Bungarus multicinctus, cobra, king snake, long-noded pit viper, pine
Flower snake, brave at least one of tail snake and water snake.
The invention also discloses a kind of methods for preparing above-mentioned snakeskin collagen peptide, comprising the following steps:
It extracts snakeskin collagen: being made after snakeskin is removed blood, degerming, decontamination, crushing, degreasing, extraction, purifying and freeze-drying
At snakeskin collagen protein freeze-dried powder；
Pre-treatment: take snakeskin collagen protein freeze-dried powder is soluble in water to obtain snakeskin collagen protein sample liquid；
Digestion: protein incision enzyme digestion is added in above-mentioned snakeskin collagen protein sample liquid and obtains digested liquid, repeats above-mentioned
Digestion step 0-3 times, after digestion, enzyme deactivation activity is cooling, adjusts the pH value of digested liquid to 6-8；
Purifying: digested liquid is purified, the snakeskin collagen peptide liquid of 100Da-20KDa is prepared；
It is dry: snakeskin collagen peptide liquid is dried to obtain snakeskin collagen peptide dried product.
Preferably, in the pre-treatment step, the mass ratio of the snakeskin collagen protein freeze-dried powder and water is 1:5-25.
Preferably, in the digestion step, protein incision enzyme include Collagenase, neutral proteinase, bromelain,
At least one of papain, pancreatin, pepsin, alkali protease, the protein incision enzyme and the snakeskin collagen
The ratio of albumen is 10000-200000U/g.
Preferably, in the digestion step, digestion condition are as follows: the pH value of snakeskin collagen protein sample liquid is adjusted to 2-11, enzyme
Cutting temperature is 10-60 DEG C, and the digestion time is 0.5-8h；
Preferably, the step of the enzyme deactivation activity are as follows: 85-95 DEG C of effect 10-60min；
Preferably, the pH value of digested liquid is adjusted using sodium hydroxide.
Preferably, it in the purification step, dialyses to obtain molecular weight using ultrafiltration membrane retention, nanofiltration membrane retention or bag filter
For the snakeskin collagen peptide liquid of 100Da-20KDa；
In the drying steps, snakeskin collagen peptide dried product is obtained using vacuum freeze drying or spray drying.
The present invention also provides the preparation method systems of a kind of above-mentioned snakeskin collagen peptide or above-mentioned snakeskin collagen peptide
Standby obtained snakeskin collagen peptide is preparing the purposes in drug, health care product or cosmetics.
The present invention also provides a kind of preparation side including above-mentioned snakeskin collagen peptide or above-mentioned snakeskin collagen peptide
The cosmetics for resisting age of skin for the snakeskin collagen peptide that method is prepared, based on parts by weight, including following raw material: octanoic acid/
5-9 parts of Triglyceride DDD, 3-7 parts of cyclomethicone, 3-7 parts of cetostearyl alcohol, 3-5 parts of PEG-100 stearate,
2-4 parts of stearine, Butyrospermum parkii (BUTYROSPERMUM PARK II) 3-5 parts of fruit rouge, Nipasol 0.05-0.15
Part, 0.1-0.3 parts of stearoyl-glutamic acid sodium, 0.1-0.3 parts of methyl hydroxybenzoate, 0.4-0.6 parts of xanthan gum, 4-8 parts of glycerol, bis- (hydroxyls
Methyl) 0.1-0.5 parts of imidazolidinyl urea, 0.1-0.3 parts of methyl hydroxybenzoate, 0.05-0.15 parts of Nipasol, snakeskin collagen peptide
0.5-2 parts and deionized water 50-75 parts.
Preferably, based on parts by weight, including following raw material: 7 parts of caprylic/capric triglyceride, ring polydimethylsiloxanes
5 parts of alkane, 5 parts of cetostearyl alcohol, 3.80 parts of PEG-100 stearate, 2.50 parts of stearine, Butyrospermum parkii
(BUTYROSPERMUM PARK II) 3.50 parts of fruit rouge, 0.10 part of Nipasol, 0.20 part of stearoyl-glutamic acid sodium, oxybenzene first
0.20 part of ester, 0.50 part of xanthan gum, 6.00 parts of glycerol, 0.30 part of Bis(hydroxymethyl)imidazolidinyl urea, 0.20 part of methyl hydroxybenzoate, hydroxyl
0.10 part of phenylpropyl alcohol ester, 1.00 parts of snakeskin collagen peptide and 64.6 parts of deionized water.
A kind of method that the present invention has carried out in also palace cosmetics for resisting age of skin for preparing above-mentioned snakeskin collagen peptide, including
Following steps:
S1, caprylic/capric triglyceride, cyclomethicone, cetostearyl alcohol are weighed according to selected parts by weight,
PEG-100 stearate, stearine, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge and Nipasol mix
Conjunction is heated to it and is completely dissolved, and obtains A phase solution；
S2, stearoyl-glutamic acid sodium, methyl hydroxybenzoate and portions of de-ionized water, Hybrid Heating are weighed according to selected parts by weight
It is completely dissolved to it, obtains B phase solution；
S3, by the A phase solution of acquisition and B phase solution mixing homogeneous, be then added into mixed solution according to selected
The weighed xanthan gum of parts by weight and glycerol, homogeneous, keep the temperature and stir, and are added when being then cooled to 50 DEG C or less according to selected weight
Measure the snake of the weighed Bis(hydroxymethyl)imidazolidinyl urea of number, methyl hydroxybenzoate, Nipasol and remainder deionized water dissolving
Collagen peptide solution, stirs evenly, be cooled to room temperature to get.
Preferably, in S1 the and S2 step, weighed raw material is mixed and heated to 80-85 DEG C.
Preferably, it in the S3 step, keeps the temperature and stirs 20-40 minutes.
Technical solution of the present invention has the advantages that
1. snakeskin collagen peptide provided by the invention, using snakeskin collagen as raw material, snakeskin has as Chinese medicine
Skin care effect, the snakeskin collagen peptide active constituent content that snakeskin collagen generates after digestion is more, and molecular weight is
100Da-20KDa, has preferable antioxygenic property, and the snakeskin collagen peptide DPPH clearance rate of 1mg/ml concentration reaches
49.36%, superoxide anion clearance rate reaches 51.12%.
2. snakeskin is removed impurity, lipid, foreign protein class by the preparation method of snakeskin collagen peptide provided by the invention,
Extracting snakeskin collagen is enzymatic hydrolysis raw material, and the resulting snakeskin collagen peptide yield of digestion is high, is all larger than 50%, and obtain
Product invalid components it is less, impurity is few, and antioxygenic property is good, and separation process is simple, with short production cycle.
3. the preparation method of snakeskin collagen peptide provided by the invention carries out compound digestion using multiple protein restriction endonuclease
It,, can be because of making on the basis of guaranteeing effective component yield due to using snakeskin collagen as raw material or when substep digestion
It is hydrolyzed with different protein incision enzymes in different sites, obtains a greater variety of snakeskin collagen peptides, inoxidizability is more
It is good.
4. the preparation method of snakeskin collagen peptide provided by the invention is retained or saturating using ultrafiltration membrane retention, nanofiltration membrane
Analysis bag dialysis purifies the snakeskin collagen peptide that these mild purification modes obtain, and prevents the anti-of damage snakeskin collagen peptide
Oxidation activity.
5. the preparation method of snakeskin collagen peptide provided by the invention, saves snakeskin collagen by the way of freeze-drying
With snakeskin collagen peptide, the holding time is long and preservation effect is good, and the activity of snakeskin collagen peptide also can be protected preferably
It holds.
6. cosmetics for resisting age of skin provided by the invention, including snakeskin collagen peptide of the present invention have and live
Property it is high, compatibility is strong, highly-safe has list, and the peptide molecule of micron level, it is easy to depth is directly entered by skin peptide surface layer
Layer skin peptide, has significant antioxidant activity, plays significant resisting age of skin effect, and preparation method mild condition, has
The activity of effect protection snakeskin collagen peptide.
Specific embodiment
There is provided following embodiments is to preferably further understand the present invention, it is not limited to the best embodiment party
Formula is not construed as limiting the contents of the present invention and protection scope, anyone under the inspiration of the present invention or by the present invention and its
The feature of his prior art is combined and any and identical or similar product of the present invention for obtaining, all falls within of the invention
Within protection scope.
Specific experiment step or condition person are not specified in embodiment, according to the literature in the art described routine experiment
The operation of step or condition can carry out.Reagents or instruments used without specified manufacturer, being can be by commercially available acquisition
Conventional reagent product.
Embodiment 1
Present embodiments provide a kind of specific embodiment of snakeskin collagen protein extraction, comprising the following steps:
Except blood, degerming: water snake snakeskin is cut into a section shape, with mass fraction be 4% NaCl solution impregnate 4h after more
The NaCl solution renewed, is repeated 4 times, and drains；
Decontamination: the NaCO for being 10% with mass fraction3Solution soaking and stirring 16h is then used pure water 3 times；
It crushes: snakeskin being lyophilized with vacuum freeze drier, is crushed to obtain snakeskin powder with pulverizer after freeze-drying；
Degreasing: n-hexane is added, degreasing is carried out to snakeskin powder, be aided with ultrasonic 2h, frequency 100KHz is subsequently agitated for
12h drains n-hexane and obtains pretreated snakeskin powder, and wherein the ratio of n-hexane and snakeskin powder is 10ml:1g；
It extracts: the snakeskin powder after degreasing being dissolved in 1mol/L citric acid solution, 4 DEG C of standing 40min.The snakeskin powder
The last ratio with the citric acid solution is 1g:10ml, continues thereafter with and 1mol/L citric acid solution is added, until the snakeskin powder
The last ratio with the citric acid solution is 1g: 60ml, extracts 1h at 105 DEG C in high-pressure steam sterilizing pan；
Purifying: after extraction, circular centrifugal 4 times at 30Hz with tube centrifuge discard snakeskin residue, collect permeate
For the thick solution of collagen；The final concentration of 5M of NaCl to NaCl is added into the thick solution of collagen, stirring keeps NaCl completely molten
Solution left standstill is for 24 hours after solution；With tube centrifuge at 30Hz circular centrifugal 4 times, centrifugation rate 18000rpm, collect on filter cloth
Albumen precipitation；Add water into albumen precipitation, promotes albumen precipitation to redissolve under 65 DEG C of water-baths, the ratio of the water and albumen precipitation that add
For 60ml:1g, ultrafiltration then is carried out with desalination to solution with 10K Da ultrafiltration membrane, collects concentrate, as collagen solution；
Freeze-drying: vacuum freeze drying, -20 DEG C of preservations are carried out to collagen solution.
Embodiment 2
Present embodiments provide a kind of specific embodiment of snakeskin collagen protein extraction, comprising the following steps:
Except blood, degerming: zaocys dhumnade snakeskin being cut into a section shape, is added after the NaCl solution immersion 4h that mass fraction is 4% more
The NaCl solution renewed is repeated 5 times, and is drained；
Decontamination: the NaCO for being 10% with mass fraction3Solution soaking and stirring 12h is then used pure water 5 times；
It crushes: snakeskin being lyophilized with vacuum freeze drier, is crushed after freeze-drying with pulverizer to get snakeskin powder；
Degreasing: n-hexane is added, degreasing is carried out to snakeskin powder, be aided with ultrasonic 3h, supersonic frequency 5OKHz is subsequently agitated for
16h drains n-hexane and obtains pretreated snakeskin powder, and wherein the ratio of n-hexane and snakeskin is 15ml:1g；
It extracts: the snakeskin powder after degreasing being dissolved in 1mol/L citric acid solution, 4 DEG C of standing 30min；The snakeskin powder
The last ratio with the citric acid solution is 1g:10ml, continues thereafter with and 1mol/L citric acid solution is added, until the snakeskin powder
The last ratio with the citric acid solution is 1g: 60ml, extracts 3h at 100 DEG C；
Purifying: after extraction, with tube centrifuge, circular centrifugal 4 times at 30Hz, centrifugation rate 18000rpm discard snake
Skin residue, collection permeate are the thick solution of collagen；Into the thick solution of collagen be added NaCl to NaCl concentration be 5M,
Stirring makes NaCl be completely dissolved rear solution left standstill for 24 hours；With tube centrifuge at 30Hz circular centrifugal 4 times, the egg on filter cloth is collected
White precipitating；Add water into albumen precipitation, promotes albumen precipitation to redissolve under 65 DEG C of water-baths, add the amount of water and the ratio of albumen precipitation
For 30ml:1g；Ultrafiltration then is carried out with desalination to solution with 10KDa ultrafiltration membrane, collects concentrate, as collagen solution；
Freeze-drying: vacuum freeze drying, -20 DEG C of preservations are carried out to collagen solution.
Embodiment 3
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
25 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 7.5, Collagenase, the Collagenase and the periostracum serpentis gelatin is added
The ratio of former protein freeze-dried powder is 150000U:1g, and digestion 2h is carried out at 48 DEG C；Enzyme deactivation is 20 minutes living at 95 DEG C, by digestion
Liquid is cooled to room temperature, and the pH value of digested liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide using sodium hydroxide solution；It will
The enzymolysis liquid is dialysed through 20KDa ultrafiltration membrane and 100Da bag filter, the snakeskin collagen peptide liquid of 100Da-20KDa is obtained, by collagen
Peptide liquid carries out vacuum freeze drying and obtains snakeskin collagen peptide dried product.
Embodiment 4
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
5 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder as described in example 2
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 7, neutral proteinase, the neutral proteinase and the snakeskin collagen is added
The ratio of protein freeze-dried powder is 100000U:1g, and digestion 8h is carried out at 10 DEG C；Enzyme deactivation is 40 minutes living at 90 DEG C, by digested liquid
It is cooled to room temperature, the pH value of digested liquid is transferred to 8 to get the enzymolysis liquid for arriving snakeskin collagen peptide using sodium hydroxide solution；By institute
It states enzymolysis liquid to dialyse through 20KDa ultrafiltration membrane and 100Da bag filter, the snakeskin collagen peptide liquid of 100Da-20KDa is obtained, by collagen peptide
Liquid carries out vacuum freeze drying and obtains snakeskin collagen peptide dried product.
Embodiment 5
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
15 times of water dissolutions, as periostracum serpentis gelatin protein sample liquid will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1；
Snakeskin collagen protein sample liquid pH value is adjusted to 2.2, pepsin is added, the pepsin and the snakeskin collagen freeze
The ratio of dry powder is 10000U:1g, and digestion 2h is carried out at 55 DEG C；Enzyme deactivation is 60 minutes living at 85 DEG C, and digested liquid is cooled to
The pH value of digested liquid is transferred to 7 using sodium hydroxide solution to get the enzymolysis liquid for arriving snakeskin collagen peptide by room temperature；By the enzymatic hydrolysis
Liquid is dialysed through 20KDa ultrafiltration membrane and 100Da bag filter, obtains the snakeskin collagen peptide liquid of 100Da-20KDa, collagen peptide liquid is carried out
Vacuum freeze drying obtains snakeskin collagen peptide dried product.
Embodiment 6
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
20 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 10.5, alkali protease, the Collagenase and the periostracum serpentis gelatin is added
The ratio of former protein freeze-dried powder is 200000U:1g, and digestion 0.5h is carried out at 60 DEG C；Enzyme deactivation is 10 minutes living at 95 DEG C, by enzyme
It cuts liquid and is cooled to room temperature, the pH value of digested liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide using sodium hydroxide solution；
The enzymolysis liquid is dialysed through 20KDa ultrafiltration membrane and 1KDa bag filter, the snakeskin collagen peptide liquid of 100Da-20KDa is obtained, by glue
Former peptide liquid carries out vacuum freeze drying and obtains snakeskin collagen peptide dried product.
Embodiment 7
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using pallas pit viper snakeskin
As raw material, snakeskin collagen peptide is prepared according to embodiment described in embodiment 3, difference is, what the present embodiment used
Protein incision enzyme is bromelain.
Embodiment 8
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using poisonless snake snake
Skin prepares snakeskin collagen peptide as raw material, according to embodiment described in embodiment 3, and difference is, the present embodiment uses
Protein incision enzyme be papain.
Embodiment 9
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using Bungarus multicinctus snake
Skin prepares snakeskin collagen peptide as raw material, according to embodiment described in embodiment 3, and difference is, the present embodiment uses
Protein incision enzyme be pancreatin.
Embodiment 10
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using glasses calmy
Skin adds 25 times of water dissolutions, as snakeskin collagen protein sample liquid as raw material, by snakeskin collagen protein freeze-dried powder obtained；It will
Snakeskin collagen protein sample liquid pH value is adjusted to 8, and Collagenase and neutral proteinase, the Collagenase and the snake is added
The ratio of collagen freeze-dried powder is 100000U:1g, the ratio of the Collagenase and the snakeskin collagen protein freeze-dried powder
Example is 50000U:1g, and digestion 2h is carried out at 48 DEG C；Enzyme deactivation is 20 minutes living at 95 DEG C, and digested liquid is cooled to room temperature, utilizes
The pH value of digested liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide by sodium hydroxide solution；By the enzymolysis liquid through 10KDa
Ultrafiltration membrane and 3KDa nanofiltration membrane obtain the snakeskin collagen peptide liquid of 3KDa-10KDa, and collagen peptide liquid progress vacuum freeze drying is obtained
To snakeskin collagen peptide dried product.
Embodiment 11
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using king calmy
Skin adds 25 times of water dissolutions, as snakeskin collagen protein sample liquid as raw material, by snakeskin collagen protein freeze-dried powder obtained；It will
Snakeskin collagen protein sample liquid pH value is adjusted to 8, and Collagenase is added, and the Collagenase and the snakeskin collagen freeze
The ratio of dry powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then neutral proteinase, the neutral proteinase is added
Ratio with the snakeskin collagen protein freeze-dried powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then at 95 DEG C
Enzyme deactivation is 20 minutes living, and digested liquid is cooled to room temperature, and the pH value of digested liquid is transferred to 7 to get snake is arrived using sodium hydroxide solution
The enzymolysis liquid of collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, the periostracum serpentis gelatin of 3KDa-10KDa is obtained
Collagen peptide liquid progress vacuum freeze drying is obtained snakeskin collagen peptide dried product by former peptide liquid.
Embodiment 12
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using long-noded pit viper snake
Skin adds 25 times of water dissolutions, as snakeskin collagen protein sample liquid as raw material, by snakeskin collagen protein freeze-dried powder obtained；It will
Snakeskin collagen protein sample liquid pH value is adjusted to 2, and pepsin, the pepsin and the snakeskin collagen protein freeze-dried powder is added
Ratio be 100000U:1g, at 48 DEG C carry out digestion 1h；Then snakeskin collagen protein sample liquid pH value is adjusted to 7, be added
The ratio of neutral proteinase, the neutral proteinase and the snakeskin collagen protein freeze-dried powder is 100000U:1g, at 48 DEG C
Carry out digestion 0.5h；Then 20 minutes living of enzyme deactivation at 95 DEG C, are cooled to room temperature for digested liquid, using sodium hydroxide solution by enzyme
The pH value for cutting liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration
Film obtains the snakeskin collagen peptide liquid of 3KDa-10KDa, and it is drying that collagen peptide liquid progress vacuum freeze drying is obtained snakeskin collagen peptide
Product.
Embodiment 13
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using preserved egg calmy
Skin adds 25 times of water dissolutions, as snakeskin collagen protein sample liquid as raw material, by snakeskin collagen protein freeze-dried powder obtained；It will
Snakeskin collagen protein sample liquid pH value is adjusted to 2, and pepsin, the pepsin and the snakeskin collagen protein freeze-dried powder is added
Ratio be 50000U:1g, at 48 DEG C carry out digestion 3h；Then snakeskin collagen protein sample liquid pH value is adjusted to 7, glue is added
The ratio of former protease, the Collagenase and the snakeskin collagen protein freeze-dried powder is 60000U:1g, is carried out at 48 DEG C
Digestion 1h；Then 20 minutes living of enzyme deactivation at 95 DEG C, are cooled to room temperature for digested liquid, using sodium hydroxide solution by digested liquid
PH value is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, obtain
Collagen peptide liquid progress vacuum freeze drying is obtained snakeskin collagen peptide dried product by the snakeskin collagen peptide liquid of 3KDa-10KDa.
Embodiment 14
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
The present embodiment prepares snakeskin collagen according to preparation method described in embodiment 1, and difference is, using brave tail calmy
Skin adds 25 times of water dissolutions, as snakeskin collagen protein sample liquid as raw material, by snakeskin collagen protein freeze-dried powder obtained；It will
Snakeskin collagen protein sample liquid pH value is adjusted to 8, and neutral proteinase is added, and the neutral proteinase and the snakeskin collagen freeze
The ratio of dry powder is 150000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then Collagenase, the Collagenase is added
Ratio with the snakeskin collagen protein freeze-dried powder is 120000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then at 95 DEG C
Enzyme deactivation is 20 minutes living, and digested liquid is cooled to room temperature, and the pH value of digested liquid is transferred to 7 to get snake is arrived using sodium hydroxide solution
The enzymolysis liquid of collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, the periostracum serpentis gelatin of 3KDa-10KDa is obtained
Collagen peptide liquid progress vacuum freeze drying is obtained snakeskin collagen peptide dried product by former peptide liquid.
Embodiment 15
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
25 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 2, pepsin, the pepsin and the snakeskin collagen is added
The ratio of freeze-dried powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then by snakeskin collagen protein sample liquid pH value tune
To 7, it being added Collagenase, the ratio of the Collagenase and the snakeskin collagen protein freeze-dried powder is 80000U:1g,
Digestion 0.5h is carried out at 48 DEG C；Then neutral proteinase, the neutral proteinase and the snakeskin collagen protein freeze-dried powder is added
Ratio be 100000U:1g, at 48 DEG C carry out digestion 0.5h；Then enzyme deactivation is 20 minutes living at 95 DEG C, and digested liquid is cooling
To room temperature, the pH value of digested liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide using sodium hydroxide solution；By the enzyme
Liquid is solved through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, obtains the snakeskin collagen peptide liquid of 3KDa-10KDa, collagen peptide liquid is subjected to vacuum
Freeze-drying obtains snakeskin collagen peptide dried product.
Embodiment 16
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
25 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 2, pepsin, the pepsin and the snakeskin collagen is added
The ratio of freeze-dried powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then by snakeskin collagen protein sample liquid pH value tune
To 7, it being added neutral proteinase, the ratio of the neutral proteinase and the snakeskin collagen protein freeze-dried powder is 80000U:1g,
Digestion 0.5h is carried out at 48 DEG C；Then Collagenase, the Collagenase and the snakeskin collagen protein freeze-dried powder is added
Ratio be 100000U:1g, at 48 DEG C carry out digestion 0.5h；Then enzyme deactivation is 20 minutes living at 95 DEG C, and digested liquid is cooling
To room temperature, the pH value of digested liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide using sodium hydroxide solution；By the enzyme
Liquid is solved through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, obtains the snakeskin collagen peptide liquid of 3KDa-10KDa, collagen peptide liquid is subjected to vacuum
Freeze-drying obtains snakeskin collagen peptide dried product.
Embodiment 17
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
25 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 2.5, pepsin, the pepsin and the snakeskin collagen egg is added
The ratio of white freeze-dried powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then by snakeskin collagen protein sample liquid pH value
7 are adjusted to, neutral proteinase and Collagenase, the ratio of the neutral proteinase and the snakeskin collagen protein freeze-dried powder is added
For 80000U:1g, the ratio of the Collagenase and the snakeskin collagen protein freeze-dried powder is 100000U:1g, at 48 DEG C
Carry out digestion 5h；Then 20 minutes living of enzyme deactivation at 95 DEG C, are cooled to room temperature for digested liquid, using sodium hydroxide solution by digestion
The pH value of liquid is transferred to 7 to get the enzymolysis liquid for arriving snakeskin collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane,
The snakeskin collagen peptide liquid of 3KDa-10KDa is obtained, collagen peptide liquid progress vacuum freeze drying is obtained into snakeskin collagen peptide dried product.
Embodiment 18
Present embodiments provide a kind of specific embodiment of snakeskin collagen peptide.
25 times of water dissolutions, as snakeskin collagen protein sample will be added by snakeskin collagen protein freeze-dried powder described in embodiment 1
Liquid；Snakeskin collagen protein sample liquid pH value is adjusted to 2, pepsin, the pepsin and the snakeskin collagen is added
The ratio of freeze-dried powder is 100000U:1g, and digestion 0.5h is carried out at 48 DEG C；Then by snakeskin collagen protein sample liquid pH value tune
To 10, it being added alkali protease, the ratio of the alkali protease and the snakeskin collagen protein freeze-dried powder is 80000U:1g,
Digestion 0.5h is carried out at 48 DEG C；Then snakeskin collagen protein sample liquid pH value is adjusted to 7, is added neutral proteinase, it is described in
Property protease and the snakeskin collagen protein freeze-dried powder ratio be 80000U:1g, at 48 DEG C carry out digestion 5h；Then 95
20 minutes living of enzyme deactivation, are cooled to room temperature for digested liquid at DEG C, using sodium hydroxide solution by the pH value of digested liquid be transferred to 7 to get
To the enzymolysis liquid of snakeskin collagen peptide；By the enzymolysis liquid through 10KDa ultrafiltration membrane and 3KDa nanofiltration membrane, the snake of 3KDa-10KDa is obtained
Collagen peptide liquid progress vacuum freeze drying is obtained snakeskin collagen peptide dried product by collagen peptide liquid.
Embodiment 19
Present embodiments provide a kind of matching comprising the cosmetics for resisting age of skin of snakeskin collagen peptide described in embodiment 3
Side and preparation method thereof is as follows:
[formula] caprylic/capric triglyceride 7g, cyclomethicone 5g, cetostearyl alcohol 5g, PEG-100 are stearic
Acid esters 3.8g, stearine 2.5g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 3.5g, Nipasol
0.1g, stearoyl-glutamic acid sodium 0.2g, methyl hydroxybenzoate 0.2g, xanthan gum 0.5g, glycerol 6g, snakeskin collagen peptide 1g, go from
Sub- water 64.6g, Bis(hydroxymethyl)imidazolidinyl urea 0.3g, methyl hydroxybenzoate 0.2g and Nipasol 0.1g
[preparation method] takes caprylic/capric triglyceride 7g, cyclomethicone 5g, cetostearyl alcohol 5g, PEG-
100 stearate 3.8g, stearine 2.5g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 3.5g, oxybenzene
Propyl ester 0.1g is heated to 80 DEG C of uniformly mixed resulting mixtures, by stearoyl-glutamic acid sodium 0.2g methyl hydroxybenzoate 0.2g and deionization
Water 10g is heated to 80 DEG C of uniformly mixed are added in said mixture and stirs evenly, and xanthan gum 0.5g and glycerol 6g is mixed equal
Even to be added in the aforementioned mixture mixed, homogeneous 10min is subsequently placed in 80 DEG C of water-baths and keeps the temperature 30min while stirring, drop
Temperature to 50 DEG C, will snakeskin collagen peptide 1g be added 54.6g deionized water in dissolve, then with Bis(hydroxymethyl)imidazolidinyl urea
0.3g, methyl hydroxybenzoate 0.2g, Nipasol 0.1g are uniformly mixed, and are added in the good mixture of aforementioned homogeneous, are stirred evenly i.e.
?.
Embodiment 20
A kind of cosmetics for resisting age of skin comprising snakeskin collagen peptide described in embodiment 13 is present embodiments provided to match
Side and preparation method thereof is as follows:
[formula] caprylic/capric triglyceride 5g, cyclomethicone 3g, cetostearyl alcohol 3g, PEG-100 are stearic
It is acid esters 3g, stearine 2g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 3g, Nipasol 0.05g, hard
Acyl sodium glutamate 0.1g, methyl hydroxybenzoate 0.1g, xanthan gum 0.4g, glycerol 4g, snakeskin collagen peptide 0.5g, deionized water
75g, then with Bis(hydroxymethyl)imidazolidinyl urea 0.1g, methyl hydroxybenzoate 0.1g and Nipasol 0.05g.
[preparation method] takes caprylic/capric triglyceride 5g, cyclomethicone 3g, cetostearyl alcohol 3g, PEG-
100 stearate 3g, stearine 2g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 3g, Nipasol
0.05g is heated to 80 DEG C of uniformly mixed resulting mixtures, by stearoyl-glutamic acid sodium 0.1g, methyl hydroxybenzoate 0.1g and deionized water
20g is heated to 80 DEG C of uniformly mixed are added in said mixture and stirs evenly, and xanthan gum 0.4g and glycerol 4g are uniformly mixed
It is added in the aforementioned mixture mixed, homogeneous 10min, is subsequently placed in 80 DEG C of water-baths and keeps the temperature 30min while stirring, cool down
To 50 DEG C, will snakeskin collagen peptide 0.5g be added 55g deionized water in dissolve, then with Bis(hydroxymethyl)imidazolidinyl urea
0.1g, methyl hydroxybenzoate 0.1g, Nipasol 0.05g are uniformly mixed, and are added in the good mixture of aforementioned homogeneous, are stirred evenly i.e.
?.
Embodiment 21
It present embodiments provides a kind of comprising the cosmetics for resisting age of skin of snakeskin collagen peptide described in embodiment 18
Formula and preparation method thereof is as follows:
[formula] caprylic/capric triglyceride 9g, cyclomethicone 7g, cetostearyl alcohol 7g, PEG-100 are stearic
It is acid esters 5g, stearine 4g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 5g, Nipasol 0.15g, hard
Acyl sodium glutamate 0.3g, methyl hydroxybenzoate 0.3g, xanthan gum 0.6g, glycerol 8g, snakeskin collagen peptide 2g, deionized water 50g,
Bis(hydroxymethyl)imidazolidinyl urea 0.5g, methyl hydroxybenzoate 0.3g and Nipasol 0.15g.
[preparation method] takes caprylic/capric triglyceride 9g, cyclomethicone 7g, cetostearyl alcohol 7g, PEG-
100 stearate 5g, stearine 4g, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge 5g, Nipasol
0.15g is heated to 80 DEG C of uniformly mixed resulting mixtures, by stearoyl-glutamic acid sodium 0.3g, methyl hydroxybenzoate 0.3g and deionized water
10g is heated to 80 DEG C of uniformly mixed are added in said mixture and stirs evenly, and xanthan gum 0.6g and glycerol 8g are uniformly mixed
It is added in the aforementioned mixture mixed, homogeneous 10min, is subsequently placed in 80 DEG C of water-baths and keeps the temperature 30min while stirring, cool down
To 50 DEG C, will snakeskin collagen peptide 2g be added 40g deionized water in dissolve, then with Bis(hydroxymethyl)imidazolidinyl urea
0.5g, methyl hydroxybenzoate 0.3g, Nipasol 0.15g are uniformly mixed, and are added in the good mixture of aforementioned homogeneous, are stirred evenly i.e.
?.
Comparative example 1
This comparative example provides a kind of specific embodiment of snakeskin collagen peptide, described in this comparative example and embodiment 3
Embodiment is essentially identical, and difference is, raw material used by this comparative example is snakeskin slurry, and preparation method includes following step
It is rapid: water snake snakeskin being cleaned with clear water, is rubbed with meat grinder, rubbing degree has the control of Φ 6mm stainless steel plate, then uses
Supermicro mill defibrination to fineness be 10-30um to obtain the final product.
Comparative example 2
The specific embodiment that this comparative example provides a kind of collagen peptide is pressed using tilapia fishskin as raw material
Rofe Isin glue collagen is prepared according to specific embodiment described in embodiment 1, according still further to specific embodiment described in embodiment 3
Prepare Rofe Isin glue collagen peptide.
Comparative example 3
This comparative example provides a kind of specific embodiment of cosmetics for resisting age of skin.
Cosmetics for resisting age of skin described in this comparative example is prepared according to embodiment described in embodiment 19, difference exists
In: this comparative example does not contain snakeskin collagen peptide.
Evaluate example 1
According to embodiment 1-18, comparative example 1, the resulting snakeskin collagen peptide dried product of comparative example 2 weight m2With make
The weight m of snakeskin collagen protein raw materials2, obtain the yield a, a=m of snakeskin collagen1/m2× 100%, such as 1 institute of table
Show.
1 embodiment 1-18 of table, comparative example 1, the yield of the resulting snakeskin collagen peptide of comparative example 2
Evaluate example 2
By 1,1- diphenyl -2- trinitrophenyl-hydrazine, i.e. DPPH is dissolved in dehydrated alcohol, and the DPPH for configuring 0.2mg/ml is female
Liquid is protected from light storage in 4 DEG C, and the DPPH ethanol solution of 0.05mg/ml is diluted to before use.
Example 3- embodiment 18 respectively, the collagen peptide dried product 4mg of comparative example 1 and comparative example 2, embodiment 19
Gained cosmetics 0.4g, cosmetics 0.8g described in embodiment 20, cosmetics 0.2g described in embodiment 21 make up described in comparative example 3
Product 0.4g is added deionized water and is settled to 2ml in test tube, adds the DPPH solution that 2mL concentration is 0.04mg/mL, mixing
Uniformly, 20min is reacted, 3500rpm is centrifugated 10min, and taking supernatant to survey its light absorption value at 517nm is Ai；
It separately respectively takes the enzymatic hydrolysis supernatant solution of the above-mentioned concentration of 2ml in test tube, is separately added into dehydrated alcohol 2ml, react
20min, 3500rpm are centrifugated 10min, take supernatant to survey its light absorption value at 517nm and are denoted as Aj；With 2mL 0.04mg/mL
DPPH and 2mL dehydrated alcohol is reacted as reference, and light absorption value is denoted as A0, by the calculating of following formula, obtain embodiment 3-
Embodiment 21, the DPPH clearance rate E (DPPH) of comparative example 1-3, as shown in table 2；
E (DPPH)=[1- (Ai-Aj)/A0] × 100%；
In formula, ask liquid to the free radical scavenging activity (%) of DPPH on E (DPPH)-enzymatic hydrolysis；
A0The light absorption value of -2ml DPPH solution addition 2ml dehydrated alcohol；
AiThe light absorption value of 2ml enzymatic hydrolysis supernatant is added in -2ml DPPH solution；
AjThe light absorption value of 2ml enzymatic hydrolysis supernatant is added in -2ml water-ethanol.
2 embodiment 3- embodiment 21 of table, the DPPH clearance rate of comparative example 1-2
| Embodiment | DPPH clearance rate (%) | Embodiment | DPPH clearance rate (%) |
| 3 | 21.23 | 14 | 40.12 |
| 4 | 22.56 | 15 | 41.02 |
| 5 | 21.63 | 16 | 49.36 |
| 6 | 23.03 | 17 | 49.36 |
| 7 | 21.59 | 18 | 48.59 |
| 8 | 23.68 | 19 | 53.68 |
| 9 | 22.58 | 20 | 55.33 |
| 10 | 21.56 | 21 | 55.26 |
| 11 | 39.65 | Comparative example 1 | 10.25 |
| 12 | 35.25 | Comparative example 2 | 12.37 |
| 13 | 38.67 | Comparative example 3 | 0.21 |
Evaluate example 3
Using assay NBT photoreduction testing example 3- embodiment 18, the superoxide anion of comparative example 1 and comparative example 2
Inhibiting rate.
Equivalent weighs embodiment 3- embodiment 18, and the collagen peptide 60mg of comparative example 1 and comparative example 2 is dissolved in 1ml's
Solution is made in deionized water, takes solution 0.1ml that Tris-HCl (pH8.2) buffer of 2.8ml 0.1mol/L is added；Blank
Control tube replaces sample with deionized water, and concussion mixes, and the pyrogallol solution of 0.1mM/L is added after 25 DEG C of water-bath 10min
(25 DEG C of water-bath preheatings) mixes and starts timing rapidly, and light absorption value is measured at 320nm, reads A320 every 30s, ties after 5min
Beam.It is returned to zero with Tris-HCl (pH8.2) buffer of 0.1mol/L, as the regression equation that absorbance changes over time, tiltedly
Rate is mouse thymus cells rate v, and sample is calculated as follows to the inhibiting rate of superoxide anion:
Inhibiting rate=(v control-v sample)/v control × 100%；
In formula, v control is control group mouse thymus cells rate, and v sample is sample sets mouse thymus cells rate；
Calculated result is as shown in table 3.
The clearance rate of the collagen peptide superoxide anion of 3 embodiment 3- embodiment 18 of table, comparative example 1 and comparative example 2
| Embodiment | Superoxide anion clearance rate (%) | Embodiment | Superoxide anion clearance rate (%) |
| 3 | 31.21 | 12 | 42.18 |
| 4 | 31.58 | 13 | 39.62 |
| 5 | 32.05 | 14 | 39.98 |
| 6 | 31.69 | 15 | 42.53 |
| 7 | 32.56 | 16 | 49.63 |
| 8 | 33.05 | 17 | 50.23 |
| 9 | 32.58 | 18 | 51.12 |
| 10 | 31.93 | Comparative example 1 | 15.2 |
| 11 | 31.59 | Comparative example 2 | 14.26 |
Obviously, the above embodiments are merely examples for clarifying the description, and does not limit the embodiments.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And it is extended from this it is obvious variation or
It changes still within the protection scope of the invention.
Claims (13)
1. a kind of snakeskin collagen peptide, which is characterized in that the snakeskin collagen peptide using snakeskin collagen prepare and
At molecular weight 100Da-20KDa.
2. snakeskin collagen peptide according to claim 1, which is characterized in that the molecular weight of the snakeskin collagen peptide
For 3KDa-10KDa；Preferably, the snakeskin is selected from zaocys dhumnade, pallas pit viper, poisonless snake, Bungarus multicinctus, cobra, king snake, five steps
Snake, preserved egg snake, brave at least one of tail snake and water snake.
3. a kind of method for preparing snakeskin collagen peptide as claimed in claim 1 or 2, which comprises the following steps:
It extracts snakeskin collagen: snake is made after snakeskin is removed blood, degerming, decontamination, crushing, degreasing, extraction, purifying and freeze-drying
Collagen freeze-dried powder；
Pre-treatment: take snakeskin collagen protein freeze-dried powder is soluble in water to obtain snakeskin collagen protein sample liquid；
Digestion: protein incision enzyme digestion is added in above-mentioned snakeskin collagen protein sample liquid and obtains digested liquid, repeats above-mentioned digestion
Step 0-3 times, after digestion, enzyme deactivation activity is cooling, adjusts the pH value of digested liquid to 6-8；
Purifying: digested liquid is purified, the snakeskin collagen peptide liquid of 100Da-20KDa is prepared；
It is dry: snakeskin collagen peptide liquid is dried to obtain snakeskin collagen peptide dried product.
4. the preparation method of snakeskin collagen peptide according to claim 3, which is characterized in that the pre-treatment step
In, the mass ratio of the snakeskin collagen protein freeze-dried powder and water is 1:5-25.
5. the preparation method of snakeskin collagen peptide according to claim 3 or 4, which is characterized in that the digestion step
In, protein incision enzyme includes Collagenase, neutral proteinase, bromelain, papain, pancreatin, pepsin, alkali
Property at least one of protease, the ratio of the protein incision enzyme and the snakeskin collagen is 10000-200000U/g.
6. according to the preparation method of the described in any item snakeskin collagen peptides of claim 3-5, which is characterized in that the digestion
In step, digestion condition are as follows: the pH value of snakeskin collagen protein sample liquid is adjusted to 2-11, and digestion temperature is 10-60 DEG C, the digestion time
For 0.5-8h；Preferably, the step of the enzyme deactivation activity are as follows: 85-95 DEG C of effect 10-60min；Preferably, using sodium hydroxide
Adjust the pH value of digested liquid.
7. according to the preparation method of the described in any item snakeskin collagen peptides of claim 3-6, which is characterized in that the purifying
In step, ultrafiltration membrane retention, nanofiltration membrane retention or bag filter is used to dialyse the snakeskin collagen for obtaining molecular weight as 100Da-20KDa
Protein peptides liquid；In the drying steps, snakeskin collagen peptide dried product is obtained using vacuum freeze drying or spray drying.
8. snakeskin collagen peptide of any of claims 1 or 2 or the described in any item snakeskin collagen peptides of claim 3-7
The snakeskin collagen peptide that is prepared of preparation method preparing the purposes in drug, health care product or cosmetics.
9. one kind includes snakeskin collagen peptide of any of claims 1 or 2 or the described in any item periostracum serpentis gelatins of claim 3-7
The cosmetics for resisting age of skin of former protein peptides, which is characterized in that based on parts by weight, including following raw material: caprylic/capric glycerol
Three 5-9 parts of esters, 3-7 parts of cyclomethicone, 3-7 parts of cetostearyl alcohol, 3-5 parts of PEG-100 stearate, glyceryl stearate
2-4 parts of acid esters, Butyrospermum parkii (BUTYROSPERMUM PARK II) 3-5 parts of fruit rouge, 0.05-0.15 parts of Nipasol, stearoyl
0.1-0.3 parts of sodium glutamate, 0.1-0.3 parts of methyl hydroxybenzoate, 0.4-0.6 parts of xanthan gum, 4-8 parts of glycerol, bis- (methylol) imidazoles
0.1-0.5 parts of ureine, 0.1-0.3 parts of methyl hydroxybenzoate, 0.05-0.15 parts of Nipasol, 0.5-2 parts of snakeskin collagen peptide and
50-75 parts of deionized water.
10. the cosmetics for resisting age of skin of snakeskin collagen peptide according to claim 9, which is characterized in that with weight
Number meter, including following raw material: 7 parts of caprylic/capric triglyceride, 5 parts of cyclomethicone, 5 parts of cetostearyl alcohol,
3.80 parts of PEG-100 stearate, 2.50 parts of stearine, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge
3.50 parts, 0.10 part of Nipasol, 0.20 part of stearoyl-glutamic acid sodium, 0.20 part of methyl hydroxybenzoate, 0.50 part of xanthan gum, glycerol
6.00 parts, 0.30 part of Bis(hydroxymethyl)imidazolidinyl urea, 0.20 part of methyl hydroxybenzoate, 0.10 part of Nipasol, snakeskin collagen
1.00 parts and 64.6 parts of deionized water of peptide.
11. a kind of method for preparing the cosmetics for resisting age of skin of snakeskin collagen peptide described in claim 10 or 9, special
Sign is, includes the following steps:
S1, caprylic/capric triglyceride, cyclomethicone, cetostearyl alcohol, PEG- are weighed according to selected parts by weight
100 stearates, stearine, Butyrospermum parkii (BUTYROSPERMUM PARK II) fruit rouge and Nipasol, mixing add
Heat is completely dissolved to it, obtains A phase solution；
S2, stearoyl-glutamic acid sodium, methyl hydroxybenzoate and portions of de-ionized water are weighed according to selected parts by weight, is mixed and heated to it
It is completely dissolved, obtains B phase solution；
S3, by the A phase solution of acquisition and B phase solution mixing homogeneous, then into mixed solution be added according to selected weight
The weighed xanthan gum of number and glycerol, homogeneous, keep the temperature and stir, and are added when being then cooled to 50 DEG C or less according to selected parts by weight
Count the periostracum serpentis gelatin of weighed Bis(hydroxymethyl)imidazolidinyl urea, methyl hydroxybenzoate, Nipasol and remainder deionized water dissolving
Former albumen peptide solution, stirs evenly, be cooled to room temperature to get.
12. the preparation method of the cosmetics for resisting age of skin of snakeskin collagen peptide according to claim 11, feature
It is, in S1 the and S2 step, weighed raw material is mixed and heated to 80-85 DEG C.
13. the preparation method of the cosmetics for resisting age of skin of snakeskin collagen peptide according to claim 11 or 12,
It is characterized in that, in the S3 step, keeps the temperature and stir 20-40 minutes.
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| CN114903176A (en) * | 2022-05-09 | 2022-08-16 | 广州博厚健康科技有限公司 | Hypoallergenic polypeptide nutrition powder and preparation method thereof |
| CN114903176B (en) * | 2022-05-09 | 2023-11-21 | 博厚健康科技股份有限公司 | Low-sensitization polypeptide nutrition powder and preparation method thereof |
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Effective date of registration: 20200416 Address after: 215500 Xinzhuang section, provincial road 227, Changshu City, Suzhou City, Jiangsu Province Applicant after: Suzhou Crowley Cosmetics Co.,Ltd. Address before: 215555 Longliqi Biological Industrial Park, Changshu City, Jiangsu Province Applicant before: Jiangsu Longrich Bioscience Co.,Ltd. |
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Address after: 215500 Xinzhuang section of 227 provincial road, Changshu City, Suzhou City, Jiangsu Province Applicant after: Crowley Cosmetics Co.,Ltd. Address before: 215500 Xinzhuang section of 227 provincial road, Changshu City, Suzhou City, Jiangsu Province Applicant before: Suzhou Crowley Cosmetics Co.,Ltd. |
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Application publication date: 20190416 |