CN106967165A - It is a kind of that the method that reducing agent extracts keratin from rabbit fur fibre is made with cysteine - Google Patents
It is a kind of that the method that reducing agent extracts keratin from rabbit fur fibre is made with cysteine Download PDFInfo
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- CN106967165A CN106967165A CN201610020960.3A CN201610020960A CN106967165A CN 106967165 A CN106967165 A CN 106967165A CN 201610020960 A CN201610020960 A CN 201610020960A CN 106967165 A CN106967165 A CN 106967165A
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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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4741—Keratin; Cytokeratin
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- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The invention discloses one kind from rabbit fur fibre method for extracting proteins, it is related to the applied technical field of rabbit hair resource.This method is that the rabbit hair mainly is hydrolyzed into 5h 75 DEG C in 8mol/l urea and 0.15mol/l cysteine mixed liquors, filters out the not molten rabbit hair, obtains rabbit hair protein hydrolyzate.Rabbit hair protein liquid is injected into bag filter and the 3d that dialysed in distilled water, change 18% PEG 20000 solution dialysis 1d, taking-up is freeze-dried to obtain rabbit hair albumen powder.The invention provides a kind of recovery rate is high, pollution-free, environmental protection keratin extracting method, finally obtains rabbit hair keratin content and be up to 91.75%.
Description
Technical field
It is a kind of method for extracting keratin in rabbit fur fibre the present invention relates to the technical field for extracting protein.
Background technology
Keratin material is native protein, with excellent biological compatibility, skin-friendly and nontoxicity,
It includes substantial amounts of hydrophilic radical.In the application performance of biological medicine, textile industry, engineering field and beauty etc.
Go out good prospect.The keratin protein that keratin is widely present in the feather, hair, nail of animal etc. belongs to outer
The structural proteins of endoderm cell, and keratin resource content is quite abundant.Keratin is under normal operation not
It is dissolved in water, diluted acid, diluted alkaline or salting liquid.These keratin can not be utilized by animal under normal circumstances, angle
Albumen is hydrolyzed to free amino acid after high temperature, high pressure and soda acid ferment treatment, and small peptide could be used in animal.
Keratin contains more cystine, and it also increases therewith containing disulfide bond.Because these disulfide bond are in protein peptide
Crosslinked action is played in chain, therefore the chemical property of keratin is highly stable, and have high mechanical properties, it is considered as tool
There is the potential high-quality protein source of higher nutritive value and steady quality.
The rabbit hair is a kind of animal origin of function admirable, with soft texture, smooth frivolous, pure white vivid, is protected
The advantages of warm property, gas permeability and preferable hygroscopicity, be extraordinary textile raw material.But in textile process process
In, due to the characteristic of rabbit fur fibre in itself, the fuds fuddled of many is occurred in that, if discarded, environment is not only polluted,
And waste the amino acid resource in the albumen of preciousness.Rabbit fur fibre contains as a kind of animal origin, protein
Amount is the good material for extracting keratin more than 90%.However, extracting the method for keratin at present has a variety of,
Wherein reducing process be effect preferably, it is the most widely used, conventional reducing agent have mercaptoethanol, TCEP,
With sodium hydrogensulfite etc., first two is has a virose reducing agent, and to carry dissolution rate relatively low for sodium hydrogensulfite, all
The problem of existing various.Therefore, find a kind of new extraction keratin method be in the urgent need to.
The content of the invention
The purpose of the present invention provides one kind aiming at above-mentioned problem and uses cysteine to be used as reducing agent
The method that method extracts keratin from the rabbit hair.By the preparation method of the present invention, rabbit hair dissolution rate is up to
74.12%, the rabbit hair keratin high purity 91.75% finally extracted.
Brief description of the drawings
The biuret reaction chromogenic reaction of Fig. 1 rabbit hair keratin
The infrared spectrogram of Fig. 2 rabbit hair keratin
Extraction process
The present invention is to use cysteine as reducing agent to extract keratin, concrete operations technique from rabbit fur fibre
It is as follows：
Pretreatment takes 5g rabbit hairs bubble, and cleaning removes a small amount of impurity contained in fiber repeatedly in warm water, after cleaning
Dried in 70 DEG C of constant temperature ovens, and be cut into the small fragment of 1cm length；
Extraction process：The pretreated rabbit hairs of 5g are placed in 40% urea liquid that bath raio is 1: 5, used
Its pH value is adjusted to hydrolyze 5h at 10,75 DEG C by NaOH solution.The undissolved rabbit fur fibre of filtering is taken out, will
Rabbit hair keratin liquor injection bag filter (molecular cut off 8000-14000) is dialysed 3 days in distilled water
Removal of impurities, a water is changed every 4h, then is placed in 18% polyglycol solution 1d and is gone out excessive moisture, and freezing is dry
It is dry to obtain keratin powder.
1. the measure of dissolution rate
The undissolved rabbit hair is cleaned multiple times with distilled water, drying weighs quality at 70 DEG C.Its dissolution rate is as follows：
Wherein, quality g before the dissolving of the M rabbit hairs；m1Undissolved rabbit hair quality g.
5g rabbit fur fibres are by after above-mentioned hydrolysis, its dissolution rate is 74.12% after measured.
2. biuret chromogenic reaction
Biuret reagent is (with 1.5g copper sulphate (CuSO4·H2) and 6.0g rocchelle's salts O
(KNaC4H4O6·4H2O), dissolved with 500ml distilled water, be added slowly with stirring 300ml10%NaOH
Solution, is diluted with water to 1L.
The biuret developer for measuring 5ml is added in test tube, adds 1ml rabbit hair protein hydrolyzate, with
The biuret solution for not adding rabbit hair keratin is blank control group, as shown in figure 1, compared by color, it is molten
Liquid is changed into purple from blueness, illustrates that with biuret reagent complexing occurs for the peptide bond that protein matter contains in solution instead
Should, generate the complex compound of aubergine.
3. the infrared spectrum analysis of rabbit hair keratin samples
It can be analyzed in characteristic frequency area by upper figure, in 3200cm-1~3400cm-1Nearby there is obvious absorption
Band, peak shape is roomy, belongs to herein in the acidic-group in protein into the ammonia in the hydroxyl and basic group of hydrogen bond
The stretching vibration of base absorbs.Importantly, in 1650cm-1It is acid amides I that extremely strong absworption peak, which occurs, in place
Band, caused by C=O key chatterings are flexible；And in 1546cm-1Equally there is more obvious vibration absorption peak at place,
As caused by the stretching vibration of C-N keys and N-H flexural vibrations, belong to acid amides II bands；Exist in addition
1250cm-1Place also have faint vibration absorption peak, belong to the characteristic absorption of acid amides III bands, be respectively from C-N,
C-O stretching vibration and O=C-N flexural vibrations.Characteristic absorption peak above is the feature official of protein
Caused by rolling into a ball.
4. Ke Daerding nitrogen
The rabbit hair keratin powder for measuring 0.1994g extraction is placed in kjeldahl flask, add 0.5g copper sulphate and
Potassium sulfate (mass ratio 3: 1) mixture and the 10ml concentrated sulfuric acids, plus 4~5 beades, and alcolhol burner slowly adds
Heat, until solution dissolving, and by black transposition water white transparency, and continue to heat about 1h, taking-up is cooled to room
Temperature.50ml distilled water is slowly added into flask, not molten salt is dissolved, cooling is transferred to 100ml volumetric flask
In, volumetric flask repeatedly is incorporated to distilled water flushing flask, add water configuration scale, shakes up.
The digestion liquid for measuring 20ml is added in steam generator, is added in 20ml40% NaOH solution and mistake
Many concentrated sulfuric acids, until solution blackening.Place the conical flask for the boric acid solution for loading 30ml5% in condenser pipe lower end
And instill 4 drop bromocresol green methyl red indicators.Heating causes reactor reaction generation ammonia and passes through condenser pipe
Flow into conical flask so that solution becomes green by pink, determine the pH value of efflux for after neutral and after
Continuous reaction 0.5h can stop.
Take out solution to be titrated with 0.11285mol/l hydrochloric acid solutions so that solution becomes aubergine by green, read
Hydrochloric acid consumption.
In formula, c：Titrate concentration of hydrochloric acid mol/l；V1, V2：Reading ml before and after hydrochloric acid solution；m：Rabbit hair keratin
The quality g of powder；
It is 14.68% to measure nitrogen content by calculating, according to the calculating of protein content
The total nitrogen content %*k of protein %=,
Wherein k is conversion factor, and k values are 6.25 in hair.
Measured by calculating containing protein content 91.75%.
Claims (5)
- Make reducing agent with cysteine and extract the method for keratin from the rabbit hair to be characterized in that and use 1. a kind of Cysteine extracts keratin as a kind of reducing agent of environmental protection from the rabbit hair, and this invention is for improving rabbit The value of hair has profound significance.
- 2. according to claims 1, keratin is extracted from the 2g rabbit hairs and is characterised by after cleaning The rabbit hair is cut into the fragment of 1cm length, adds the 100ml urea containing 8mol/l and 0.15mol/l half Guang In the solution of propylhomoserin.
- 3. according to claims 2, the above-mentioned solution for soaking the rabbit hair is placed in heating water bath at 75 DEG C 5h, taking-up filters out the not molten rabbit hair, obtains rabbit hair protein liquid.
- 4. according to claims 3, the rabbit hair protein liquid of collection is injected in bag filter, distilled water is placed After 3~5d, 18% PEG 20000 solution 1d is placed into until most solutions dialysis is fallen.
- 5. according to claims 4, obtained concentrate egg white liquor is placed in freeze drier and dried, finally Obtain the rabbit hair keratin powder of white.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610020960.3A CN106967165A (en) | 2016-01-13 | 2016-01-13 | It is a kind of that the method that reducing agent extracts keratin from rabbit fur fibre is made with cysteine |
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| CN201610020960.3A CN106967165A (en) | 2016-01-13 | 2016-01-13 | It is a kind of that the method that reducing agent extracts keratin from rabbit fur fibre is made with cysteine |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107982551A (en) * | 2017-11-21 | 2018-05-04 | 东华大学 | The method that biomineralization method prepares gadolinium oxide base magnetic resonance contrast agent |
| CN108498446A (en) * | 2018-07-11 | 2018-09-07 | 佛山皖阳生物科技有限公司 | A kind of hair conditioner and preparation method thereof for preventing hair quality impaired |
| CN109575119A (en) * | 2018-12-25 | 2019-04-05 | 广州美炫化妆品有限公司 | The Novel extraction method of rabbit hair keratin and its application in cosmetics |
| CN110373892A (en) * | 2019-07-02 | 2019-10-25 | 天津工业大学 | Modified rabbit fur fibre of a kind of graphene and preparation method thereof |
| CN111592588A (en) * | 2020-04-26 | 2020-08-28 | 中梁农业股份有限公司 | Extraction system of rabbit hair keratin |
| CN114887123A (en) * | 2022-04-18 | 2022-08-12 | 南通大学 | Hirudin grafted nanofiber vascular stent material, preparation method and application |
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| CN101020715A (en) * | 2007-03-15 | 2007-08-22 | 王利忠 | Process of extracting and preparing deer nerve growth factor (DEER NGF) |
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| CN101020715A (en) * | 2007-03-15 | 2007-08-22 | 王利忠 | Process of extracting and preparing deer nerve growth factor (DEER NGF) |
| CN104861663A (en) * | 2015-04-28 | 2015-08-26 | 东华大学 | Method for regenerating wool keratin by L-cysteine |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107982551A (en) * | 2017-11-21 | 2018-05-04 | 东华大学 | The method that biomineralization method prepares gadolinium oxide base magnetic resonance contrast agent |
| CN108498446A (en) * | 2018-07-11 | 2018-09-07 | 佛山皖阳生物科技有限公司 | A kind of hair conditioner and preparation method thereof for preventing hair quality impaired |
| CN109575119A (en) * | 2018-12-25 | 2019-04-05 | 广州美炫化妆品有限公司 | The Novel extraction method of rabbit hair keratin and its application in cosmetics |
| CN110373892A (en) * | 2019-07-02 | 2019-10-25 | 天津工业大学 | Modified rabbit fur fibre of a kind of graphene and preparation method thereof |
| CN111592588A (en) * | 2020-04-26 | 2020-08-28 | 中梁农业股份有限公司 | Extraction system of rabbit hair keratin |
| CN114887123A (en) * | 2022-04-18 | 2022-08-12 | 南通大学 | Hirudin grafted nanofiber vascular stent material, preparation method and application |
| CN114887123B (en) * | 2022-04-18 | 2023-07-04 | 南通大学 | Hirudin grafted nanofiber vascular stent material, preparation method and application |
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