CN102229665A - Method for extracting poultry feather keratin by using water insoluble ionic liquid and relative poultry feather keratin solution - Google Patents
Method for extracting poultry feather keratin by using water insoluble ionic liquid and relative poultry feather keratin solution Download PDFInfo
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Abstract
The invention provides a method for extracting a poultry feather keratin by using water insoluble ionic liquid, comprising the follow steps: 1) washing, drying and crushing a poultry feather; 2) adding a water insoluble ionic liquid with stirring for 3-6h at the temperature of 70-100 DEG C; and 3) after the reaction, adding water, carrying out centrifugation to form a three-phase system of water insoluble ionic liquid/ undissolved poultry feather/ water, so as to obtain a crude keratin solution. In a preferred step 2), a disulfide bond reducing agent is added, and a little water is added to dissolve the disulfide bond reducing agent. The invention also provides a poultry feather keratin solution extracted by the method. The method for extracting the poultry feather keratin by using the water insoluble ionic liquid, has the advantage of simple separation of keratin, is convenient recovery of ionic liquid and friendly to environmental protection, and can be widely applied.
Description
Technical field
The present invention relates to poultry feather Keratin sulfate extractive technique field, be meant a kind of keratic method of water-insoluble ionic liquid extract poultry feather and relevant poultry feather keratin solution of adopting especially.
Background technology
Keratin sulfate is the bioprotein that the source is abundant, using value is very high, extensively is present in the weave construction of organism.The main chemical compositions of feather is a Keratin sulfate, as a kind of green natural fiber, has the irreplaceable advantage of other materials.The annual poultry feather output in the whole world can reach several hundred million tons, and the feather waste output of China reaches more than 70 ten thousand tons, but also is increasing year by year.Utilize waste feathers to prepare the solubility feather protein and not only made full use of protein resource, also significantly reduced the environmental pollution that causes thus simultaneously.China payes attention to the recycling of feather refuse since about the eighties in 20th century, and for example, the Keratin sulfate hydrolysate is as the additive of animal-feed, and its modified product is filled leather, rubber etc., but processability excellent leather and rubber product.The Keratin sulfate of making not only can be used for spinning, also can be follow-up the application in fields such as food, chemical industry, medicine, agricultural, sewage disposal, packing, building, archaeology, weavings and lays the first stone, and produces good economic and social benefit.
The keratin molecule chain is made of 19 kinds of a-amino acids, and amino acid constitutes the polypeptide long-chain by peptide bond, and these long-chains form keratic space structure by lateral ties such as disulfide linkage, hydrogen bond, sat linkage, Van der Waals forces again.Generally speaking, the disulfide linkage in the keratin molecule makes and takes place crosslinkedly between peptide chain inside and the peptide chain, forms tridimensional network, makes natural keratin be difficult to dissolving, gives that keratic extraction brings difficulty in the feather.Extracting keratic method at present roughly has mechanical process, chemical method and biological process etc., and wherein chemical method can be divided into acid and alkali hydrolysis method, reduction method, oxidation style etc. again.Chinese patent 200810150653.2, " extracting keratic method from feather " disclosed, this invention is to extract Keratin sulfate with chemical method from feather, and its main technique flow process is: alcohol pre-treatment → hydrochloric acid pre-treatment → mercapto-ethanol reduction → Keratin sulfate precipitation.The technical finesse steps such as chemical method of existing report are many, and utilize the unfriendly reagent of environment such as a large amount of inorganic acid alkalis and reductive agent, cause very big environmental pollution.
Ionic liquid is by organic cation and the salt that is liquid state under near temperature room temperature or the room temperature inorganic or that organic anion constitutes.Compare with traditional organic solvent, ionic liquid has following characteristics: (1) liquid state temperature range wide (2) steam forces down, not volatile (3) all show good dissolving ability (4) to a large amount of inorganic and organic substances and have bigger designability, and density is big.Because ionic liquid shows these unique physicochemical property and peculiar function, so it is novel " soft " functional materials or medium that a class is worth researchdevelopment.Present ion liquid Application Areas mainly exists: chemical reaction, catalyzed reaction, isolation technique, electrochemistry etc.Phillips in 2004 etc. have at first studied the dissolution characteristics of different ionic liquid to silk, found that hydrophilic imidazoles villaumite extracts silk fiber solute effect (J.Am.Chem.Soc., 2004,126 (44): 14350-14351) are preferably arranged.Thank to hypo etc. and studied and utilize hydrophilic imidazoles villaumite to prepare the method for keratin solution, obtain solute effect (CN200410011389.6) preferably.By extensive studies and utilization, but its cost is higher relatively as a kind of green solvent for ionic liquid, so ionic liquid must be recycled.Though above-mentioned hydrophilic ionic liquids has solute effect preferably to silk, feather etc., keratic separation and ion liquid recovery need be added organic solvent, have caused the raising of cost and the pollution of environment.
Take a broad view of above-mentioned prior art, traditional method is extracted keratic defective and is: reagent dosages such as inorganic acid alkali are big, and can not recycle, and environment is caused great pollution; The keratic defective of ionic liquid extract is: Keratin sulfate is difficult to separate from ionic liquid.Therefore, utilize ion liquid designability, seeking a kind of suitable ion liquid dissolving feather becomes this area problem demanding prompt solution.
Summary of the invention
Main purpose of the present invention is exactly the problems and shortcomings at above existence, a kind of keratic method of water-insoluble ionic liquid extract poultry feather and relevant poultry feather keratin solution of adopting is provided, adopt this extracting method, Keratin sulfate separates simple, ionic liquid reclaims convenient, and environmental friendliness is suitable for large-scale promotion application.
In order to solve above-mentioned purpose, in a first aspect of the present invention, provide a kind of employing water-insoluble ionic liquid extract poultry feather keratic method, be characterized in that described method may further comprise the steps:
1) with poultry feather washing, dry, pulverizing;
2) add the water-insoluble ionic liquid, stirred 3～6 hours down at 70 ℃～100 ℃;
3) add water, centrifugal back formation water-insoluble ionic liquid/do not dissolve poultry feather/water three-phase system after the reaction, isolate water, promptly obtain thick keratin solution.
Described water-insoluble ionic liquid can be any water-fast ionic liquid.So-called " water insoluble " is meant that the solubleness of this ionic liquid in water is less than 8g/100mL.Preferably, described water-insoluble ionic liquid is two fluoroform sulfimide salt [HOEMIm] NTf of 1-hydroxyethyl-3-Methylimidazole
2
Described poultry feather and the ion liquid mass ratio of described water-insoluble can be any suitable ratios without limits.Preferably, described poultry feather and the ion liquid mass ratio of described water-insoluble are 1: 20～1: 50.
Though ionic liquid can destroy disulfide linkage, in order to promote the destruction of disulfide linkage, preferably, in described step 2) in, also add the disulfide bond reduction agent, and less water is to dissolve described disulfide bond reduction agent.The disulfide bond reduction agent more can promote the destruction of disulfide linkage.
Because it is to pulverize poultry feather and be in order fully to contact, so its degree of grinding without limits, but the smaller the better with ionic liquid and disulfide bond reduction agent; In addition, step 2) add less water in and be in order to dissolve described disulfide bond reduction agent, amount is few more good more.
Described disulfide bond reduction agent can be any disulfide bond reduction agent.Preferably, described disulfide bond reduction agent is sodium bisulfite (NaHSO
3).
The mass ratio of described poultry feather and described disulfide bond reduction agent can be any suitable ratio without limits.Preferably, the mass ratio of described poultry feather and described disulfide bond reduction agent is 1: 0.3～1: 1.5.
In order to make full use of the water-insoluble ionic liquid and not dissolve poultry feather, preferably, described method also comprises:
31) after isolating water, in the insoluble ionic liquid of remainder water, there is the undissolved poultry feather of part, adds the poultry feather of drying and crushing in addition, and add the water-insoluble ionic liquid, stirred 3～6 hours down at 70 ℃～100 ℃;
32) carry out step 3).
In order to make full use of the water-insoluble ionic liquid more and not dissolve poultry feather, above-mentioned steps 31) and 32) can constantly repeat, therefore, preferably, described method also comprises:
33) repeating step 31) and 32).
In order to obtain purer keratin solution, preferably, described method also comprises:
4) described thick keratin solution is obtained the purified keratin solution through the dialysis desalting purifying.
In a second aspect of the present invention, a kind of poultry feather keratin solution is provided, be characterized in, obtain by the keratic method extraction of above-mentioned employing water-insoluble ionic liquid extract poultry feather.
Beneficial effect of the present invention is: the keratic method of employing water-insoluble ionic liquid extract poultry feather of the present invention, stirred 3～6 hours down at 70 ℃～100 ℃ by poultry feather and the water-insoluble ionic liquid that will pulverize, add water, centrifugal back then and form water-insoluble ionic liquid/do not dissolve poultry feather/water three-phase system, isolate water, promptly obtain thick keratin solution, reclaimed the water-insoluble ionic liquid simultaneously, Keratin sulfate separates simple, ionic liquid reclaims convenient, and environmental friendliness is suitable for large-scale promotion application.
Embodiment
In order more to be expressly understood content of the present invention, describe in detail especially exemplified by following examples.All adopt infrared spectra, SDS-polyacrylamide gel electrophoresis that keratic structure, the molecular weight that extracts characterized in the following example:
1, Infrared spectroscopy
Adopt the Infrared spectrum scanning instrument to measure the keratic infrared spectra that the present invention extracts, the line data analysis of going forward side by side is at 3080cm
-1Near polypeptide is arranged (charateristic avsorption band CO-NH-) is at 1651cm
-1, 1542cm
-1, 1236cm
-1Three absorption peaks at place are respectively acid amides I, the II of keratin molecule, the charateristic avsorption band of III band.As can be seen, there is the absorption of proteinic characteristic group's peptide bond in the Keratin sulfate.
2, SDS-polyacrylamide gel electrophoresis
Adopt SDS-polyacrylamide gel electrophoresis technology that the keratic molecular weight distribution of extracting is investigated, find that the keratin molecule amount mainly is distributed in 14kDa～29kDa.
Embodiment 1 ionic liquid [HOEMIm] NTf
2The mensuration of solubleness in water
1) ion liquid full wavelength scanner
Accurately take by weighing 50mg ionic liquid [HOEMIm] NTf
2(Lanzhou materialization institute of the Chinese Academy of Sciences, purity＞99%) places volumetric flask, adds the deionized water constant volume, is made into the 1.0mg/mL standardized solution.With deionized water is that blank reference carries out full wavelength scanner in the 190-300nm wavelength region, and can obtain this ion liquid maximum absorption band is 217.5nm.
2) mensuration of ionic liquid typical curve
Get 7 test tubes respectively, add 1.0mg/mL ionic liquid [HOEMIm] NTf that has prepared
2Solution 0,0.1,0.2,0.3,0.4,0.5 and 0.6mL, each effective deionized water add to cumulative volume be 5mL, obtain liquid to be measured.With the deionized water is reference, under maximum absorption wavelength 217.5nm, and the absorbancy of sequentially determining standard ionomer liquid sample, and drawing standard curve.
3) mensuration of ionic liquid solubleness in water
Accurately take by weighing 1.0g ionic liquid [HOEMIm] NTf
2Place the centrifuge shield of 15mL, add the 2.5mL deionized water then, on turbine mixer, stir and mixed centrifugal at last (4000r/min) 15 minutes 30 minutes.The last phase solution of getting 10 μ L with liquid-transfering gun is diluted to 25mL, and the absorbancy of test sample product is compared with typical curve, and calculating the solubleness of ionic liquid in water is 7.941g/100mL.
Embodiment 2～11 adopts the ionic liquid extract chicken feather keratin
The basic procedure that embodiment 2～11 adopts is as follows:
1) takes by weighing the chicken feather that 1g pulverizes and place round-bottomed flask, to water-insoluble ionic liquid [the HOEMIM] [NTf that wherein adds certain mass
2], sodium bisulfite and 500 μ L deionized waters, at 70 ℃～100 ℃ following mechanical stirring reaction 3～6h;
2) after reaction finishes, at first add proper amount of deionized water in reaction mixture, centrifugal back formation water-insoluble ionic liquid/do not dissolve chicken feather/water three-phase system is isolated water; Use the deionized water wash container then three times, the isolating water of washings that obtains and centrifugal back filters together, obtains thick keratin solution; The rate of recovery of last water-insoluble ionic liquid (containing a spot of chicken feather that do not dissolve) can get with drying to constant weight;
3) crude protein solution obtains purer keratin solution through dialysis (Mw6000～8000), utilizes the Xylene Brilliant Cyanine G method to survey protein soln concentration, calculates protein yields;
4) to step 2) in remaining reaction system, add chicken feather, sodium bisulfite and 500 μ L deionized waters that 1G pulverizes, add a spot of water-insoluble ionic liquid again, at 70 ℃～90 ℃ following mechanical stirring reaction 3～6h;
5) repeating step 2), 3), 4) five times.
Among the present invention, protein yields=w (protein of extraction)/w (chicken feather of adding) * 100%.
The difference of embodiment 2～11 is:
Embodiment 2: add 20g water-insoluble ionic liquid, 0.3g sodium bisulfite, add about 1.0g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 10mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 12.65%, 12.56%, 12.60%, 12.57%, 12.48%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 3: add 20g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 1.0g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 10mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 16.23%, 16.43%, 16.35%, 16.28%, 16.37%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 4: add 20g water-insoluble ionic liquid, 1.5g sodium bisulfite, add about 1.0g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 10mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 17.01%, 17.08%, 17.04%, 17.09%, 17.00%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 5: add 40g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.0g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 20mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 21.58%, 21.14%, 22.19%, 20.91%, 21.51%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 6: add 50g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.5g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 25mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 21.23%, 21.53%, 21.37%, 21.24%, 21.29%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 7: add 40g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.0g water-insoluble ionic liquid, 70 ℃ are reacted 4h down, add the Keratin sulfate of 20mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 14.65%, 14.83%, 14.77%, 14.68%, 14.73%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 8: add 40g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.0g water-insoluble ionic liquid, 100 ℃ are reacted 4h down, add the Keratin sulfate of 20mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 19.07%, 19.31%, 19.20%, 19.14%, 19.16%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 9: add 40g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.0g water-insoluble ionic liquid, 80 ℃ are reacted 3h down, add the Keratin sulfate of 20mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 17.07%, 17.21%, 17.15%, 17.22%, 17.18%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 10: add 40g water-insoluble ionic liquid, 1.0g sodium bisulfite, add about 2.0g water-insoluble ionic liquid, 80 ℃ are reacted 6h down, add the Keratin sulfate of 20mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 16.70%, 16.77%, 16.74%, 16.82%, 16.59%, and the ion liquid rate of recovery can reach about 95% at every turn.
Embodiment 11: add 20g water-insoluble ionic liquid, do not add sodium bisulfite, add about 1.0g water-insoluble ionic liquid, 80 ℃ are reacted 4h down, add the Keratin sulfate of 10mL deionized water separation and Extraction.As calculated, the Keratin sulfate productive rate that present embodiment extracts is respectively 7.86%, 7.92%, 7.88%, 7.83%, 7.80%, and the ion liquid rate of recovery can reach about 95% at every turn.
As seen, when not adding the disulfide bond reduction agent, can extract Keratin sulfate equally, and the ion liquid rate of recovery can reach about 95% at every turn, compare but add the disulfide bond reduction agent with embodiment 2, proteinic extraction yield is lower, illustrates that the disulfide bond reduction agent can obviously promote the destruction of disulfide linkage.
Therefore, the present invention extracts Keratin sulfate by adopting ionic liquid a kind of and that water is immiscible from poultry feather, extracting keratic chemism is by the interaction force in the ion liquid hyperpolarity destruction feather keratin, as hydrogen bond, sat linkage, disulfide linkage etc., promote the fracture of disulfide linkage again with the disulfide bond reduction agent, finally make feather fully be swelling to dissolving, ionic liquid is water insoluble because the Keratin sulfate that extracts is soluble in water, therefore, add entry after the dissolving and just can reach keratic separation and ion liquid recycling, obtain the Keratin sulfate aqueous solution.Need to prove that because the similarity of poultry feather, the keratic method of employing water-insoluble ionic liquid extract poultry feather of the present invention is not limited to extracts the poultry feather Keratin sulfate, obviously also is suitable for extracting other poultry feather Keratin sulfate.
In sum, adopt the keratic method of employing water-insoluble ionic liquid extract poultry feather of the present invention, Keratin sulfate separates simple, and ionic liquid reclaims convenient, and environmental friendliness, is suitable for large-scale promotion application.
In this specification sheets, the present invention is described with reference to its certain embodiments.But, still can make various modifications and conversion obviously and not deviate from the spirit and scope of the present invention.Therefore, specification sheets is regarded in an illustrative, rather than a restrictive.
Claims (9)
1. one kind is adopted the keratic method of water-insoluble ionic liquid extract poultry feather, it is characterized in that described method may further comprise the steps:
1) with poultry feather washing, dry, pulverizing;
2) add the water-insoluble ionic liquid, stirred 3～6 hours down at 70 ℃～100 ℃;
3) add water after the reaction, centrifugal back formation water-insoluble ionic liquid/do not dissolve poultry feather/water three-phase system is isolated water, promptly obtains thick keratin solution.
2. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 1 is characterized in that, described water-insoluble ionic liquid is the two fluoroform sulfimide salt of 1-hydroxyethyl-3-Methylimidazole.
3. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 1 is characterized in that described poultry feather and the ion liquid mass ratio of described water-insoluble are 1: 20～1: 50.
4. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 1 is characterized in that, in described step 2) in, also add disulfide bond reduction agent sodium bisulfite, and less water is to dissolve described disulfide bond reduction agent.
5. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 4 is characterized in that, described poultry feather and described disulfide bond reduction agent mass ratio are 1: 0.3～1: 1.5.
6. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 1 is characterized in that described method also comprises:
31) after isolating water, in the insoluble ionic liquid of remainder water, there is the not consoluet poultry feather of part, adds the poultry feather of drying and crushing in addition, and add the water-insoluble ionic liquid, stirred 3～6 hours down at 70 ℃～100 ℃;
32) carry out step 3).
7. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 6 is characterized in that described method also comprises:
33) repeating step 31) and 32).
8. the keratic method of employing water-insoluble ionic liquid extract poultry feather according to claim 1 is characterized in that described method also comprises:
4) described thick keratin solution is obtained the purified keratin solution through the dialysis desalting purifying.
9. a poultry feather keratin solution is characterized in that, obtains by extracting according to the keratic method of the arbitrary described employing water-insoluble ionic liquid extract poultry feather of claim 1～8.
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| CN104774254A (en) * | 2015-03-31 | 2015-07-15 | 上海大学 | Method for extracting keratin in feather by using imidazole chloride ionic liquid |
| CN109289734A (en) * | 2018-10-24 | 2019-02-01 | 四川建筑职业技术学院 | A kind of method and apparatus being made degradable adhesives of feather stem |
| CN109592953A (en) * | 2018-10-29 | 2019-04-09 | 吴伟华 | A kind of preparation method of freeze proof environmental protection brick |
| CN109944071A (en) * | 2017-12-20 | 2019-06-28 | 财团法人纺织产业综合研究所 | Moisture absorption heating agent and preparation method thereof |
| CN110869446A (en) * | 2017-06-02 | 2020-03-06 | 独立行政法人国立高等专门学校机构 | Molded protein, process for producing the same, protein solution, and plasticizer for molded protein |
| CN112813694A (en) * | 2021-02-01 | 2021-05-18 | 江苏康乃馨羽绒制品科技有限公司 | Fine and soft processing method of rough sheet based on fluid medium |
| CN113604529A (en) * | 2021-08-13 | 2021-11-05 | 广东完美生命健康科技研究院有限公司 | Chicken feather oligopeptide and extraction method and application thereof |
| CN117598397A (en) * | 2023-11-14 | 2024-02-27 | 安徽希普生物科技有限公司 | Preparation method and application of feather powder |
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2011
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774254A (en) * | 2015-03-31 | 2015-07-15 | 上海大学 | Method for extracting keratin in feather by using imidazole chloride ionic liquid |
| CN110869446A (en) * | 2017-06-02 | 2020-03-06 | 独立行政法人国立高等专门学校机构 | Molded protein, process for producing the same, protein solution, and plasticizer for molded protein |
| CN109944071A (en) * | 2017-12-20 | 2019-06-28 | 财团法人纺织产业综合研究所 | Moisture absorption heating agent and preparation method thereof |
| CN109289734A (en) * | 2018-10-24 | 2019-02-01 | 四川建筑职业技术学院 | A kind of method and apparatus being made degradable adhesives of feather stem |
| CN109592953A (en) * | 2018-10-29 | 2019-04-09 | 吴伟华 | A kind of preparation method of freeze proof environmental protection brick |
| CN112813694A (en) * | 2021-02-01 | 2021-05-18 | 江苏康乃馨羽绒制品科技有限公司 | Fine and soft processing method of rough sheet based on fluid medium |
| CN112813694B (en) * | 2021-02-01 | 2023-02-28 | 江苏康乃馨羽绒制品科技有限公司 | Fine and soft processing method of rough sheet based on fluid medium |
| CN113604529A (en) * | 2021-08-13 | 2021-11-05 | 广东完美生命健康科技研究院有限公司 | Chicken feather oligopeptide and extraction method and application thereof |
| CN113604529B (en) * | 2021-08-13 | 2023-10-10 | 广东完美生命健康科技研究院有限公司 | Chicken feather oligopeptide and extraction method and application thereof |
| CN117598397A (en) * | 2023-11-14 | 2024-02-27 | 安徽希普生物科技有限公司 | Preparation method and application of feather powder |
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