CN105646696A - Cowhair keratin powder and preparation method thereof - Google Patents
Cowhair keratin powder and preparation method thereof Download PDFInfo
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- CN105646696A CN105646696A CN201610022702.9A CN201610022702A CN105646696A CN 105646696 A CN105646696 A CN 105646696A CN 201610022702 A CN201610022702 A CN 201610022702A CN 105646696 A CN105646696 A CN 105646696A
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- hair
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- keratin
- cowhair
- keratin powder
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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
Abstract
The invention discloses cowhair keratin powder and a preparation method thereof. The method comprises the following steps that 1, cowhair is taken and pretreated through a surface active agent and a dilute acid solution in sequence; 2, a hydrogen peroxide alkaline solution is used for pre-hydrolyzing and bleaching the cowhair; 3, a metal salt-reducing agent combining method is used for fully hydrolyzing the cowhair; 4, then filtering, dialyzing, purifying, pH adjusting to an isoelectric point, centrifuging, drying and smashing are carried out to obtain the cowhair keratin powder. The extracting rate of cowhair keratin extracted through the method is larger than or equal to 80%, the obtained cowhair keratin powder is white, the grain size is smaller than or equal to 0.2 mm, molecular weight distribution ranges from 5 kDa to 35 kDa, and the pH ranges from 6.0 to 7.0. The cowhair keratin powder can be used for preparing a feed additive, protein filler special for leather making and other biological auxiliaries.
Description
Technical field
The invention belongs to tanning waste technical field of resource utilization. It is specifically related to ox hair keratin powder and preparation method thereof.
Background technology
In recent years, due to the continuous popularization of clean leather-making technology, in order to reduce pollution, reducing cost, hair saving process obtains promotion and application gradually. Bao Maofa the float reduced in waste water and organic while, reduce BOD and COD. Hair also non-immediate is sloughed by the protease that uses in Bao Maofa, but only clears up Rhizoma Imperatae and epidermis and do not damage hair shaft, thus can be recovered to complete hair. In order to make full use of hair produced by hair saving process, it is to avoid become the garbage in tanning production, can pass through reclaim and comprehensively utilize the purpose reaching to turn waste into wealth simultaneously, meet the thought of recycling economy.
Keratin is of many uses, and in feedstuff and food industry, keratin can as high-quality feed additive, but keratic slightly solubility influences whether animal feeding results, so that degraded hair obtains water-soluble keratin or polypeptide. Keratin is as a kind of protein fibre, it it is the bioabsorbable polymer material of a kind of function admirable, it is mainly composed of hard alpha-Keratin, this is the structure with suitable toughness and stability, make it have the advantages such as certain mechanical property, machinability and sterilizability, be applied to medical material field. In curriery, crust leather can be filled with keratin, it is possible to significantly improve richness and the percentage elongation of crust leather, and do not produce to significantly affect to the absorbance of the tensile strength of crust leather, tearing strength and dyestuff. In cosmetics and other industry: keratin hydrolysis liquid as a kind of new type natural cosmetic material, is successfully applied in the production of the cosmetics such as hair washing and skin protection by some countries. Utilize feather keratin to prepare palmityl peptide, the wetting agent that can be used for the cosmetics such as cleansing milk, protective skin cream, lipstick can be made further.
Keratin content in ox hair is more than 80%, and alpha-Keratin is protein main in hair. The aminoacid sequence of alpha-Keratin subunit is formed by rich in the central rod district of �� spiral and the unspiralized region of both sides. The �� spiral of adjacent molecule be by their cysteine residues between disulfide bond crosslinking get up, it is considered that every 4 bung flanges just have a cross-bond. The stability of ��helix, is mainly ensured by these disulfide bond. Generally, the disulfide bond in keratin molecule makes peptide chain internal and cross-links between peptide chain, forms space network, makes natural keratin be difficult to dissolve, bring difficulty to extraction keratic in hair.It addition, between keratin molecule or intramolecular hydrophobic effect, hydrogen bond, Van der Waals force, sat linkage, disulfide bond all can pass through chemical reagent be weakened or be ruptured. The wherein hydrophobic interaction of hydrophobic group in hydrogen bond, Van der Waals force, sat linkage and keratin, it is also possible to make it destroy by protein denaturant or weaken.
Ox hair can be produced certain impact by acid, alkali, oxidant, reducing agent, slaine and surfactant. But single acid, alkali, oxidant, reducing agent, slaine and surfactant but can not reach desirably to extract keratic effect. Acid-base method is to extract keratic traditional method, but can not use strong acid, highly basic, and condition difficulty controls, and excessive hydrolysis can generate aminoacid, but acidity, basicity can not be too little, otherwise can be hydrolyzed not exclusively. Single Oxidizing and Reducing Agents consumption is big, and ceratin of animal hairs extraction ratio is not high. Therefore combined techniques extracts keratin and has become as the focus of keratin hydrolysis technique study.
Summary of the invention
Big in order to solve single animal wool method for hydrolysis drug dosage, the shortcomings such as extraction ratio is low, the invention discloses a kind of ox hair keratin powder and preparation method thereof, take ox hair successively through surfactant and dilute acid soln pretreatment, ox hair is carried out prehydrolysis and bleaching by hydrogen peroxide alkaline solution, ox hair is fully hydrolyzed by slaine-reducing agent combined techniques, then filtration, dialysis, purification, adjustment pH to isoelectric point, IP, be centrifuged, dry, grind and obtain ox hair keratin powder. Adopting this method to extract ox hair keratin extraction ratio >=80%, gained ox hair keratin powder is white powder, molecular weight distribution: 5-35kDa, pH:6.0-7.0.
Ox hair keratin powder and preparation method thereof comprises the following steps:
(1) a certain amount of ox hair is weighed, sort out impurity, clean up with tap water, being 1% surfactant solution effect 1h by concentration again, ox hair quality and dosage of surfactant are than for 1g:4ml, cleaning up with tap water, the hydrochloric acid solution of concentration 1% soaks 12h, ox hair quality and hydrochloric acid solution amount ratio are amount ratio is 1g:4ml, washing, dries;
(2) being impregnated in by the ox hair that (1) processed in the hydrogen peroxide alkaline solution of pH10.0, the mass ratio of ox hair and hydrogen peroxide alkaline solution is 1:30, and at 40 DEG C of temperature, water-bath vibration 30min, clean by clean water, dries;
(3) ox hair that (2) processed being immersed in slaine with the blended liquid of reducing agent, the mass ratio of ox hair and blended liquid is water purification bath vibration at 1:30,60-70 DEG C, until ox hair is completely dissolved;
(4) ox hair lysate is taken, being cooled to room temperature, with four layer of 200 order filter-cloth filtering, filtrate moves in the bag filter of 3000 molecular weight, dialysis 48h, regulating ox hair lysate pH to keratin isoelectric point, IP 4.6, keratin precipitates out precipitation, stands 30min, centrifugal, remove supernatant, 40 DEG C of vacuum dryings, grind and obtain ox hair egg albumen powder;
Percent involved in ox hair keratin powder and preparation method thereof is mass fraction.
Wherein, the surfactant in step (1) be in dodecylbenzene sodium sulfonate, Span 20, Span 60, polysorbate40 any one, two or three.
Wherein, the hydrogen peroxide alkaline solution in step (2), compound method is, with the buffer of alkali or strong base-weak acid salt solution preparation pH10.0, and is configured to the hydrogen peroxide alkaline solution containing hydrogen peroxide concentration 10% with pH10.0 buffer and hydrogen peroxide.
Wherein, the alkali in step (2) hydrogen peroxide alkaline solution or strong base-weak acid salt be the one in sodium hydroxide, sodium acetate, disodium hydrogen phosphate, ammonia, sodium carbonate, two or three.
Wherein, the blended liquid in step (3) is formed with concentration 5%-15% reductant solution volume ratio 1:1 is blended by concentration 15%-40% metal salt solution.
Wherein, step (3) slaine is any one in zinc chloride, lithium bromide.
Wherein, the reducing agent in step (3) be in TGA, mercaptoethanol, sodium thiosulfate, sodium sulfite, sodium sulfite in one, two or three.
Detailed description of the invention
Below by enforcement, the present invention is specifically described; be necessary it is pointed out here that be that the present embodiment is served only for the present invention is further described; and it is not intended that limiting the scope of the invention, the person skilled in the art in this field can make some nonessential improvement and adjustment according to the content of foregoing invention.
Embodiment 1
Step one: weigh 100g ox hair, sorts out impurity, after tap water cleans up, is 1% Span 20 solution 400ml effect 1h by concentration, after tap water cleans up, soaks 12h with the hydrochloric acid solution 400ml of concentration 1%, and water is cleaned, and dries;
Step 2: with H2O2-NaOH solution, i.e. regulate distilled water pH to 10.0 by the NaOH solution that concentration is 2%, and configure H2O2Concentration is the H of 10%2O2-NaOH solution 300ml, takes the ox hair 5g that step one processed, is immersed in H2O2In-NaOH solution 150g, water-bath vibration 30min at 40 DEG C of temperature, ox hair is white and fully swelling, and washing is clean, dries;
Step 3: with ZnCl2-NaHSO3Solution 200ml, i.e. concentration is the ZnCl of 15%2It is 5%NaHSO with concentration3Blended with volume ratio 1:1, take the ox hair dried in step 2, water-bath vibration at adding blended liquid 150g, 60-70 DEG C, 60min ox hair is completely dissolved;
Step 4: by ox hair hydrolyzed solution, be cooled to room temperature, with four layer of 200 order filter-cloth filtering, filtrate moves in the bag filter of 3000 molecular weight, dialysis 48h, changes liquid once in every 8 hours, takes out the ox hair hydrolyzed solution after dialysis, regulating hydrolyzed solution pH value with the acetum that concentration is 1% is 4.6, keratin precipitates out precipitation, stands 30min, centrifugal, removing supernatant, 40 DEG C vacuum drying to ox hair egg albumen powder.
Adopting said method to extract keratin, keratin extraction ratio is 81%, divides in amount distribution: 5-15kDa, outward appearance is the white powder of particle diameter��0.2mm, pH:6.5.
Embodiment 2
Step one: weigh 100g ox hair, sorts out impurity, after tap water cleans up, with the dodecylbenzene sodium sulfonate solution 400ml effect 1h that concentration is 1%, after tap water cleans up, soaks 12h with the hydrochloric acid solution 400ml of concentration 1%, and water is cleaned, and dries;
Step 2: with H2O2-CH3COONa solution, i.e. be the CH of 5% by concentration3COONa solution regulates deionized water pH to 10.0, and configures H2O2Concentration is the H of 10%2O2-CH3COONa solution 300ml, takes the ox hair 5g that step one processed, is immersed in H2O2-CH3In COONa solution 150g, water-bath vibration 30min at 40 DEG C of temperature, ox hair is white and fully swelling, and washing is clean, dries;
Step 3: with LiBr-NaHSO3Solution 200ml, i.e. concentration is the LiBr of 20% is 5%NaHSO with concentration3Blended with volume ratio 1:1, take the ox hair dried in step 2, water-bath vibration at adding blended liquid 150g, 60-70 DEG C, 90min ox hair is completely dissolved;
Step 4: by ox hair hydrolyzed solution, be cooled to room temperature, with four layer of 200 order filter-cloth filtering, filtrate moves in the bag filter of 3000 molecular weight, dialysis 48h, changes liquid once in every 8 hours, takes out the ox hair hydrolyzed solution after dialysis, regulating hydrolyzed solution pH value with the acetum that concentration is 1% is 4.6, keratin precipitates out precipitation, stands 30min, centrifugal, removing supernatant, 40 DEG C vacuum drying to ox hair egg albumen powder.
Adopting said method to extract keratin, keratin extraction ratio is 80%, divides in amount distribution: 10-30kDa, outward appearance is the white powder of particle diameter��0.2mm, pH:6.7.
Embodiment 3
Step one: weigh 100g ox hair, sorts out impurity, after tap water cleans up, with the dodecylbenzene sodium sulfonate solution 400ml effect 1h that concentration is 1%, after tap water cleans up, soaks 12h with the hydrochloric acid solution 400ml of concentration 1%, and water is cleaned, and dries;
Step 2: with H2O2-NaOH solution, i.e. regulate distilled water pH to 10.0 by the NaOH solution that concentration is 2%, and configure H2O2Concentration is the H of 10%2O2-NaOH solution 300ml, takes the ox hair 5g that step one processed, is immersed in H2O2In-NaOH solution 150g, water-bath vibration 30min at 40 DEG C of temperature, ox hair is white and fully swelling, and washing is clean, dries;
Step 3: with ZnCl2-Na2S2O3Solution 200ml, i.e. concentration is the ZnCl of 20%2It is 10%Na with concentration2S2O3Blended with volume ratio 1:1, take the ox hair dried in step 2, water-bath vibration at adding blended liquid 150g, 60-70 DEG C, 90min ox hair is completely dissolved;
Step 4: by ox hair hydrolyzed solution, be cooled to room temperature, with four layer of 200 order filter-cloth filtering, filtrate moves in the bag filter of 3000 molecular weight, dialysis 48h, changes liquid once in every 8 hours, takes out the ox hair hydrolyzed solution after dialysis, regulating hydrolyzed solution pH value with the acetum that concentration is 1% is 4.6, keratin precipitates out precipitation, stands 30min, centrifugal, removing supernatant, 40 DEG C vacuum drying to ox hair egg albumen powder.
Adopting said method to extract keratin, keratin extraction ratio is 84%, divides in amount distribution: 10-20kDa, outward appearance is the white powder of particle diameter��0.2mm, pH:6.2.
Claims (9)
1. ox hair keratin powder, is characterised by, its preparation method comprises the following steps:
(1) a certain amount of ox hair is weighed, sort out impurity, clean up with tap water, again with the surfactant solution effect 1h that concentration is 1%, ox hair quality and dosage of surfactant are than for 1g:4ml, cleaning up with tap water, the hydrochloric acid solution of concentration 1% soaks 12h, ox hair quality and hydrochloric acid solution amount ratio are amount ratio is 1g:4ml, washing, dries;
(2) being impregnated in by the ox hair that (1) processed in the hydrogen peroxide alkaline solution of pH10.0, the mass ratio of ox hair and hydrogen peroxide alkaline solution is 1:30, and at 40 DEG C of temperature, water-bath vibration 30min, uses clear water wash clean, dry;
(3) ox hair that (2) processed being immersed in slaine with the blended liquid of reducing agent, the mass ratio of ox hair and blended liquid is water-bath vibration at 1:30,60-70 DEG C of temperature, until ox hair is completely dissolved;
(4) ox hair lysate is taken, being cooled to room temperature, with four layer of 200 order filter-cloth filtering, filtrate moves in the bag filter of 3000 molecular weight, dialysis 48h, regulating ox hair lysate pH to keratin isoelectric point, IP 4.6, keratin precipitates out precipitation, stands 30min, centrifugal, remove supernatant, 40 DEG C of vacuum dryings, grind and obtain ox hair egg albumen powder;
Percent involved in the preparation method of ox hair keratin powder is mass fraction.
2. the preparation method of the ox hair keratin powder described in claim 1, is characterised by, the surfactant in its step (1) be in dodecylbenzene sodium sulfonate, Span 20, Span 60, polysorbate40 any one, two or three.
3. the preparation method of the ox hair keratin powder described in claim 1, it is characterised by, hydrogen peroxide alkaline solution in its step (2), compound method is, with the buffer of alkali or strong base-weak acid salt solution preparation pH10.0, and it is configured to the hydrogen peroxide alkaline solution containing hydrogen peroxide concentration 10% with pH10.0 buffer and hydrogen peroxide.
4. the preparation method of the ox hair keratin powder described in claim 1, is characterised by, the blended liquid in its step (3) is formed with concentration 5%-15% reductant solution volume ratio 1:1 is blended by concentration 15%-40% metal salt solution.
5. the preparation method of the ox hair keratin powder described in claim 3, is characterised by, alkali in its hydrogen peroxide alkaline solution or strong base-weak acid salt be the one in sodium hydroxide, sodium acetate, disodium hydrogen phosphate, ammonia, sodium carbonate, two or three.
6. the preparation method of the ox hair keratin powder described in claim 4, is characterised by, its slaine is any one in zinc chloride, lithium bromide.
7. the preparation method of the ox hair keratin powder described in claim 1, is characterised by, the reducing agent in its step (3) be in TGA, mercaptoethanol, sodium thiosulfate, sodium sulfite, sodium sulfite in one, two or three.
8. the preparation method of the ox hair keratin powder described in claim 1-7, is characterised by, its extraction ratio is high, >=80%.
9. ox hair keratin powder described in claim 1-8 and preparation method thereof, obtained ox hair keratin powder key index:
(1) outward appearance is white powder;
(2) particle diameter :��0.2mm
(3) molecular weight distribution is wide: 5-35kDa;
(4) pH:6.0-7.0.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367544A (en) * | 2016-08-30 | 2017-02-01 | 兴业皮革科技股份有限公司 | Method for preparing compound protein filler with amino resin modified tanning waste cowhair keratin |
| CN108404200A (en) * | 2018-03-28 | 2018-08-17 | 齐鲁工业大学 | A kind of preparation method of magnetic Nano ox hair keratin sponge |
| CN111793726A (en) * | 2020-07-23 | 2020-10-20 | 嘉兴学院 | Method for preparing leather retanning agent by taking waste animal hair as raw material |
| CN113307860A (en) * | 2021-06-11 | 2021-08-27 | 四川大学 | Method for extracting keratin from cow hair recovered from tanning based on biological enzymolysis |
| CN113981610A (en) * | 2021-01-28 | 2022-01-28 | 浙江技立新材料股份有限公司 | Antibacterial keratin nanofiber membrane and preparation method thereof |
| CN117129366A (en) * | 2023-10-25 | 2023-11-28 | 武汉大学 | Method for testing high molecular weight protein extraction rate in protein extract |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367544A (en) * | 2016-08-30 | 2017-02-01 | 兴业皮革科技股份有限公司 | Method for preparing compound protein filler with amino resin modified tanning waste cowhair keratin |
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| CN108404200A (en) * | 2018-03-28 | 2018-08-17 | 齐鲁工业大学 | A kind of preparation method of magnetic Nano ox hair keratin sponge |
| CN108404200B (en) * | 2018-03-28 | 2020-12-25 | 齐鲁工业大学 | Preparation method of magnetic nano-bovine hair keratin sponge |
| CN111793726A (en) * | 2020-07-23 | 2020-10-20 | 嘉兴学院 | Method for preparing leather retanning agent by taking waste animal hair as raw material |
| CN113981610A (en) * | 2021-01-28 | 2022-01-28 | 浙江技立新材料股份有限公司 | Antibacterial keratin nanofiber membrane and preparation method thereof |
| CN113307860A (en) * | 2021-06-11 | 2021-08-27 | 四川大学 | Method for extracting keratin from cow hair recovered from tanning based on biological enzymolysis |
| CN117129366A (en) * | 2023-10-25 | 2023-11-28 | 武汉大学 | Method for testing high molecular weight protein extraction rate in protein extract |
| CN117129366B (en) * | 2023-10-25 | 2024-01-16 | 武汉大学 | Method for testing high molecular weight protein extraction rate in protein extract |
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Application publication date: 20160608 |