CN107216376A - A kind of extracting method of tussah silk fibroin - Google Patents
A kind of extracting method of tussah silk fibroin Download PDFInfo
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- CN107216376A CN107216376A CN201710626822.4A CN201710626822A CN107216376A CN 107216376 A CN107216376 A CN 107216376A CN 201710626822 A CN201710626822 A CN 201710626822A CN 107216376 A CN107216376 A CN 107216376A
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- tussah silk
- silk fibroin
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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
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
- C07K14/43586—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects from silkworms
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Abstract
Field is extracted the present invention relates to fibroin albumen, a kind of extracting method of tussah silk fibroin is disclosed, by tussah silk through degumming drying and processing；Using FeCl2Tussah silk fibroin is extracted with the sodium mixed solution system of iminodisuccinic acid four, pH is adjusted in extraction process and using ultrasonic wave added to improve recovery rate；Ultrafiltration is carried out to the solution after ultrasonically treated, surplus solution is freeze-dried after ultrafiltration, gained tussah silk fibroin clayed into power standby.The method that the present invention is provided is simple to operate, small to environmental hazard, and protein extracting ratio is high, and the fibroin albumen purity of acquisition is higher.
Description
Technical field
Field, more particularly to a kind of extracting method of tussah silk fibroin are extracted the present invention relates to fibroin albumen.
Background technology
Tussah silk is mainly made up of fibroin albumen and sericin, wherein the content of fibroin albumen about 70%, is the master of silk
Want composition.Silk gum and fibroin have different structures and performance：Silk gum is globular preteins, contains a large amount of polarity in composition amino acid
Hydrophilic side base, stability is poor, dissolves in the solution such as hot water, acid, alkali, protease and low concentration sodium carbonate；And fibroin is fibre
Shape albumen is tieed up, natural fibroin fiber contains a large amount of intramoleculars and intermolecular hydrogen bonding, and crystallinity is dissolved in common solvent compared with Gao Ernan.
Silk gum is enclosed in fibroin fiber top layer, plays adhesive action.Silk gum and fibroin have different structures and performance：Silk gum is spherical egg
In vain, secondary structure is based on random coil, and stability is poor, dissolves in hot water, acid, alkali, protease and low concentration sodium carbonate etc.
In solution；Fibroin is fibrous protein, containing substantial amounts of crystalline texture, and performance is stable, dissolves in high calcium chloride concentration, calcium chloride
In the solution such as ethanol water mixed liquid and lithium bromide.Therefore, it can according to dissolving sex differernce of the fibroin in different solutions and
Practical application request, selects suitable lysate to carry out extraction process, obtains the various silk eggs with different structure and performance
In vain, cosmetics and food additives, biological tissue's repair materials, high-absorbent material, pharmaceutical carrier and cell are prepared for processing
The different shape functional material such as culture medium.
Current existing method frequently with strong acid, highly basic, high concentration salting liquid such as high concentration CaCl2Solution, cuprammonium are molten
Liquid, lithium-bromide solution, thiocyanation lithium solution etc. dissolve to tussah silk fibroin albumen, but use the method for the above to extract silkworm
Silk-fibroin is not often thorough, it is impossible to extract the fibroin in silk cocoon completely, so as to cause the waste of material, therefore
A kind of method that can extract fibroin in silk cocoon completely as far as possible of exploitation is needed, the utilization rate of raw material is improved.
The content of the invention
In order to solve the above-mentioned technical problem, the invention provides a kind of extracting method of tussah silk fibroin.The present invention is carried
The method of confession prepares fibroin process simply, and the recovery rate of protein is high.
The present invention concrete technical scheme be：A kind of extracting method of tussah silk fibroin, comprises the following steps：
A）Weigh tussah silk and by it with 1:45-55 bath raio is added in deionized water, with salt acid for adjusting pH to 2.5-3.0,
Heated at constant temperature 55-65min carries out degumming in 98-102 DEG C of water-bath, then adds NaHCO3PH is adjusted to neutrality, by above-mentioned side
Method at least repeats degumming once；Tussah silk after degumming is added 1.5-2.5h is soaked into deionized water, then at 55-65 DEG C
Lower drying, is made tussah silk fibroin sample.
B）9-11 g FeCl2 are weighed, the imino-diacetic that 14-16 mL are added dropwise after 48-52mL deionized water dissolvings again is added
The sodium of butanedioic acid four, is settled to 100 mL, obtains mixed solution after being sufficiently mixed.
The present invention takes FeCl2Have than existing methods with the sodium mixing match solution of iminodisuccinic acid four following excellent
Point：Easily biological-degradable, it is nontoxic to the ecosystem；The sodium of iminodisuccinic acid four forces reagent to metal ion chelating capacity
It is more excellent to the extraction effect of protein.
C）By step A）Gained tussah silk fibroin sample is added to step B by solid-to-liquid ratio 0.9-1.1g/15mL）Gained is mixed
Close in solution, in 55-65 DEG C of stirred in water bath insulation 55-65min.
D）Take out after cooling, solution is placed under ultrasonic wave and handled, while it is 8.5-9.0 that pH is slowly adjusted with acetic acid；
PH value of solution is adjusted to neutrality after ultrasonically treated end.
The present invention, using ultrasonically treated, just has extraction yield higher and production in extraction process compared with conventional method
Cycle shorter advantage.
E）Solution after neutralization is subjected to ultrafiltration with ultrafiltration apparatus, it is dry to carry out freezing to the surplus solution obtained after ultrafiltration
Tussah silk fibroin that is dry, being purified, clays into power standby.
The present invention protein solution is purified using ultrafiltration apparatus, more existing dialysis process have it is simple to operate,
The short advantage of required time.
Preferably, step E）In, the ultrafiltration apparatus is poly (ether-sulfone) ultrafiltration membrane.
Preferably, the poly (ether-sulfone) ultrafiltration membrane interception is 5000.
Preferably, step D) in, ultrasonic frequency is 30-50kHz, and sonication treatment time is 25-35min.
It is compared with the prior art, the beneficial effects of the invention are as follows：
1st, FeCl is taken2With the sodium mixing match solution system of iminodisuccinic acid four, its easily biological-degradable, to the ecosystem without
Poison is harmless and strong to Protein Extraction ability.
2nd, just have extraction yield higher and the production cycle compared with conventional method using ultrasonically treated in extraction process
Shorter advantage.
3rd, protein solution is purified using ultrafiltration apparatus, more existing dialysis process have it is simple to operate, it is required
Time short advantage.
4th, the method extraction process that the present invention is provided is simple, and extracting method environmental protection, the recovery rate of fibroin is high, can reduce
The waste of fibroin resource.
Embodiment
With reference to embodiment, the invention will be further described.
Embodiment one
A 10 g tussah silks) are weighed with 1:50 bath raio is added in deionized water, with salt acid for adjusting pH to 2.5, under 95 DEG C of water-baths
Constant temperature 60min carries out degumming, and water-bath adds NaHCO after terminating3PH is adjusted to neutrality, is operated 3 times repeatedly.Tussah silk after degumming
Addition soaks 2h into 200 mL deionized waters, is then dried at 60 DEG C.
B 10 g FeCl) are weighed2, the sodium of iminodisuccinic acid four that 15 mL are added dropwise after 50mL deionized water dissolvings is added,
100 mL are settled to after being sufficiently mixed.
C 1 g steps A) is weighed）Silk sample after middle gained drying, adds 15 mL steps B）In with mixed solution,
60 min of stirring insulation in 60 DEG C of water-baths.
D) take out after cooling, solution is placed under 30 kHz ultrasonic wave and handles 30min, while slowly being adjusted with acetic acid
PH is 8.5, adjusts pH value of solution to neutrality after ultrasonically treated end.
E the solution after) neutralizing carries out ultrafiltration with ultrafiltration apparatus, and selected poly (ether-sulfone) ultrafiltration membrane interception is 5000, right
The surplus solution obtained after ultrafiltration is freeze-dried, the tussah silk fibroin purified, is clayed into power standby.
Embodiment two
A 10 g tussah silks) are weighed with 1:50 bath raio is added in deionized water, with salt acid for adjusting pH to 2.7, under 95 DEG C of water-baths
Constant temperature 60min carries out degumming, and water-bath adds NaHCO after terminating3PH is adjusted to neutrality, is operated 3 times repeatedly.Tussah silk after degumming
Addition soaks 2h into 200 mL deionized waters, is then dried at 60 DEG C.
B 10 g FeCl) are weighed2, the sodium of iminodisuccinic acid four that 15 mL are added dropwise after 50mL deionized water dissolvings is added,
100 mL are settled to after being sufficiently mixed.
C 1 g steps A) is weighed）Silk sample after middle gained drying, adds 15 mL steps B）In with mixed solution,
60 min of stirring insulation in 60 DEG C of water-baths.
D) take out after cooling, 30min is handled under the ultrasonic wave that solution is placed in 40kHz, while slowly being adjusted with acetic acid
PH is 8.8, adjusts pH value of solution to neutrality after ultrasonically treated end.
E the solution after) neutralizing carries out ultrafiltration with ultrafiltration apparatus, and selected poly (ether-sulfone) ultrafiltration membrane interception is 5000, right
The surplus solution obtained after ultrafiltration is freeze-dried, the tussah silk fibroin purified, is clayed into power standby.
Embodiment three
A 10 g tussah silks) are weighed with 1:50 bath raio is added in deionized water, and PH to 3 is adjusted with hydrochloric acid, in permanent under 95 DEG C of water-baths
Warm 60min carries out degumming, and water-bath adds NaHCO after terminating3PH is adjusted to neutrality, is operated 3 times repeatedly.Tussah silk after degumming adds
Enter and soak 2h into 200 mL deionized waters, then dried at 60 DEG C.
B 10 g FeCl) are weighed2, the sodium of iminodisuccinic acid four that 15 mL are added dropwise after 50mL deionized water dissolvings is added,
100 mL are settled to after being sufficiently mixed.
C 1 g steps A) is weighed）Silk sample after middle gained drying, adds 15 mL steps B）In with mixed solution,
60 min of stirring insulation in 60 DEG C of water-baths.
D) take out after cooling, 30min is handled under the ultrasonic wave that solution is placed in 50kHz, while slowly being adjusted with acetic acid
PH is 9, adjusts pH value of solution to neutrality after ultrasonically treated end.
E the solution after) neutralizing carries out ultrafiltration with ultrafiltration apparatus, and selected poly (ether-sulfone) ultrafiltration membrane interception is 5000, right
The surplus solution obtained after ultrafiltration is freeze-dried, the tussah silk fibroin purified, is clayed into power standby.
The present invention extracts obtained tussah silk fibroin recovery rate more than 78%.
Raw materials used in the present invention, equipment, is the conventional raw material, equipment of this area unless otherwise noted；In the present invention
Method therefor, is the conventional method of this area unless otherwise noted.
It is described above, only it is presently preferred embodiments of the present invention, not the present invention is imposed any restrictions, it is every according to the present invention
Any simple modification, change and equivalent transformation that technical spirit is made to above example, still fall within the technology of the present invention side
The protection domain of case.
Claims (4)
1. a kind of extracting method of tussah silk fibroin, it is characterised in that comprise the following steps：
A）Weigh tussah silk and by it with 1:45-55 bath raio is added in deionized water, with salt acid for adjusting pH to 2.5-3.0,
Heated at constant temperature 55-65min carries out degumming in 98-102 DEG C of water-bath, then adds NaHCO3PH is adjusted to neutrality, by above-mentioned side
Method at least repeats degumming once；Tussah silk after degumming is added 1.5-2.5h is soaked into deionized water, then at 55-65 DEG C
Lower drying, is made tussah silk fibroin sample；
B）Weigh 9-11 g FeCl2, add the iminodisuccinic acid that 14-16 mL are added dropwise after 48-52mL deionized water dissolvings again
Four sodium, are settled to 100 mL, obtain mixed solution after being sufficiently mixed；
C）By step A）Gained tussah silk fibroin sample is added to step B by solid-to-liquid ratio 0.9-1.1g/15mL）Gained mixing is molten
In liquid, in 55-65 DEG C of stirred in water bath insulation 55-65min；
D）Take out after cooling, solution is placed under ultrasonic wave and handled, while it is 8.5-9.0 that pH is slowly adjusted with acetic acid；Ultrasound
Processing is adjusted pH value of solution to neutrality after terminating；
E）Solution after neutralization is subjected to ultrafiltration with ultrafiltration apparatus, the surplus solution obtained after ultrafiltration is freeze-dried, obtained
To the tussah silk fibroin of purifying, clay into power standby.
2. a kind of extracting method of tussah silk fibroin as claimed in claim 1, it is characterised in that step E）In, it is described super
Filter device is poly (ether-sulfone) ultrafiltration membrane.
3. a kind of extracting method of tussah silk fibroin as claimed in claim 2, it is characterised in that the poly (ether-sulfone) ultrafiltration membrane
Interception is 5000.
4. a kind of extracting method of tussah silk fibroin as claimed in claim 1, it is characterised in that step D) in, ultrasonic wave
Frequency is 30-50kHz, and sonication treatment time is 25-35min.
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| CN201710626822.4A CN107216376A (en) | 2017-07-28 | 2017-07-28 | A kind of extracting method of tussah silk fibroin |
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| CN201710626822.4A CN107216376A (en) | 2017-07-28 | 2017-07-28 | A kind of extracting method of tussah silk fibroin |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109187935A (en) * | 2018-09-11 | 2019-01-11 | 浙江理工大学 | A method of Ancient Silk Textile is detected based on microwave |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1230548A (en) * | 1998-12-22 | 1999-10-06 | 金明官 | Catalytic circular preparation of fibroin peptide |
| CN104818535A (en) * | 2014-06-04 | 2015-08-05 | 大连应江柞蚕科技服务有限公司 | Unwinding method and unwinding tank capable of preserving tussah cocoon shell sericin |
| CN105017382A (en) * | 2015-08-14 | 2015-11-04 | 海安县南莫制丝有限公司 | Method for preparing soluble silk protein solution through waste silk |
| CN106435765A (en) * | 2016-12-12 | 2017-02-22 | 广西江缘茧丝绸有限公司 | Silk degumming method |
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2017
- 2017-07-28 CN CN201710626822.4A patent/CN107216376A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1230548A (en) * | 1998-12-22 | 1999-10-06 | 金明官 | Catalytic circular preparation of fibroin peptide |
| CN104818535A (en) * | 2014-06-04 | 2015-08-05 | 大连应江柞蚕科技服务有限公司 | Unwinding method and unwinding tank capable of preserving tussah cocoon shell sericin |
| CN105017382A (en) * | 2015-08-14 | 2015-11-04 | 海安县南莫制丝有限公司 | Method for preparing soluble silk protein solution through waste silk |
| CN106435765A (en) * | 2016-12-12 | 2017-02-22 | 广西江缘茧丝绸有限公司 | Silk degumming method |
Non-Patent Citations (4)
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| （美）肯•盖泽: "《无害化学品 可持续发展的策略》", 31 January 2017 * |
| 上海纺织工业专科学校: "《练漂》", 31 March 1983 * |
| 吴洁梅等: "柞蚕丝素粉的制取", 《安徽农业大学学报》 * |
| 陆旋: "pH条件对二种家蚕丝素蛋白铁元素螯合量的影响", 《安徽化工》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109187935A (en) * | 2018-09-11 | 2019-01-11 | 浙江理工大学 | A method of Ancient Silk Textile is detected based on microwave |
| CN109187935B (en) * | 2018-09-11 | 2020-12-18 | 浙江理工大学 | Method for detecting ancient silk fabric based on microwave |
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Application publication date: 20170929 |