CN107652354A - A kind of spider silk hydrolysate and preparation method thereof - Google Patents
A kind of spider silk hydrolysate and preparation method thereof Download PDFInfo
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- CN107652354A CN107652354A CN201711090220.8A CN201711090220A CN107652354A CN 107652354 A CN107652354 A CN 107652354A CN 201711090220 A CN201711090220 A CN 201711090220A CN 107652354 A CN107652354 A CN 107652354A
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- 229920001872 Spider silk Polymers 0.000 title claims abstract description 116
- 239000000413 hydrolysate Substances 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 81
- 238000005119 centrifugation Methods 0.000 claims abstract description 23
- 238000005374 membrane filtration Methods 0.000 claims abstract description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 40
- 230000007062 hydrolysis Effects 0.000 claims description 31
- 238000006460 hydrolysis reaction Methods 0.000 claims description 31
- 239000003153 chemical reaction reagent Substances 0.000 claims description 7
- 230000033228 biological regulation Effects 0.000 claims description 6
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 25
- 150000001413 amino acids Chemical class 0.000 abstract description 24
- 238000005516 engineering process Methods 0.000 abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 4
- 229920001184 polypeptide Polymers 0.000 abstract description 3
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 abstract description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 abstract description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 abstract description 2
- 238000005903 acid hydrolysis reaction Methods 0.000 abstract description 2
- 229930182817 methionine Natural products 0.000 abstract description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 abstract description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 abstract description 2
- 230000007721 medicinal effect Effects 0.000 abstract 1
- 238000000034 method Methods 0.000 description 45
- 230000008569 process Effects 0.000 description 30
- 239000011521 glass Substances 0.000 description 27
- 241000239290 Araneae Species 0.000 description 25
- 230000000052 comparative effect Effects 0.000 description 24
- 239000012535 impurity Substances 0.000 description 11
- 108010022355 Fibroins Proteins 0.000 description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 10
- 238000010298 pulverizing process Methods 0.000 description 10
- 238000001914 filtration Methods 0.000 description 8
- 238000006386 neutralization reaction Methods 0.000 description 8
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 238000001556 precipitation Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 3
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 2
- 206010013786 Dry skin Diseases 0.000 description 2
- 208000000260 Warts Diseases 0.000 description 2
- 238000003287 bathing Methods 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 210000003298 dental enamel Anatomy 0.000 description 2
- 230000037336 dry skin Effects 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000003020 moisturizing effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 201000010153 skin papilloma Diseases 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- 241001517312 Calanthe Species 0.000 description 1
- 241000486627 Chilobrachys guangxiensis Species 0.000 description 1
- 208000034507 Haematemesis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 240000005373 Panax quinquefolius Species 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 208000004680 Rectal Fistula Diseases 0.000 description 1
- 208000010340 Sleep Deprivation Diseases 0.000 description 1
- 206010051373 Wound haemorrhage Diseases 0.000 description 1
- JAWMENYCRQKKJY-UHFFFAOYSA-N [3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-ylmethyl)-1-oxa-2,8-diazaspiro[4.5]dec-2-en-8-yl]-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]methanone Chemical compound N1N=NC=2CN(CCC=21)CC1=NOC2(C1)CCN(CC2)C(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F JAWMENYCRQKKJY-UHFFFAOYSA-N 0.000 description 1
- 206010002156 anal fistula Diseases 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- CDQSJQSWAWPGKG-UHFFFAOYSA-N butane-1,1-diol Chemical compound CCCC(O)O CDQSJQSWAWPGKG-UHFFFAOYSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- -1 example 1-8 amino acid Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 108091005899 fibrous proteins Proteins 0.000 description 1
- 102000034240 fibrous proteins Human genes 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 208000014617 hemorrhoid Diseases 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 229920006253 high performance fiber Polymers 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000010415 tropism Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Peptides Or Proteins (AREA)
- Cosmetics (AREA)
Abstract
The present invention relates to amino acid preparing technical field, more particularly to a kind of spider silk hydrolysate and preparation method thereof.The spider silk hydrolysate is made using following steps：Spider silk is crushed, the spider silk after crushing is mixed with sodium hydroxide solution, and 4~5h is hydrolyzed under the conditions of 80~90 DEG C, centrifugation, obtains hydrolyzate；Hydrolyzate pH value is adjusted to 4.0~5.5,200nm membrane filtrations, obtains spider silk hydrolysate.Preparation technology acid hydrolysis yield of the present invention is 74.15%~80.89%, yield ＞ 70%；In hydrolyzate, methionine does not detect, in addition to phenylalanine, tyrosine, the equal ＞ 60% of yield of other amino acid.The amino acid in amino acids distribution and spider silk i.e. in hydrolyzate is consistent substantially.Hydrolyzate middle-molecular-weihydroxyethyl in 1000 5000D micromolecule polypeptide accounting 72.18%~76.69%, illustrate spider silk hydrolyzate both maintained it is original be used for replenishing water and preserving moisture, repair, beautifying medical effect such as crease-resistant of compacting, while it is easily absorbed by the body.
Description
Technical field
The present invention relates to amino acid preparing technical field, more particularly to a kind of spider silk hydrolysate and preparation method thereof.
Background technology
Dry skin refers to that skin lacks moisture content and makes us the phenomenon that does not feel like oneself, and its symptom is mainly skin-bound, indivedual
Dry itch all over after decortication, bathing in position.Age growth, climate change, sleep insufficiency, fatigue, water for bathing overheat,
Strong wait of articles for washing alkalescence is all the major reason for causing dry skin.Keeping the method for skin moisturizing mainly has：Skin surface
Grease is covered, reduces moisture of skin evaporation；Adequate water is provided to skin；Improve skin-nourishing situation, allow skin to recover water conservation
Function.Replenishing water and preserving moisture facial mask can provide sufficient moisture to skin, directly stick it on the face during use, persistently offer skin water
Point and nutrient.The moisturizing ingredients such as hyaluronic acid that MASQUE APAISANT HYDRATANT includes, butanediol, biological carbohydrate gum, it is very good with skin affinity,
Moisture can be provided to continuous skin.
Fibroin is the fibrous proteins of silk or the spider's thread, rich in the amino acid for forming fibroin, easily by skin
Skin absorbs, nutrition skin, and niacinamide improves skin quality.The fibroin MASQUE APAISANT HYDRATANT reported at present is the fibroin in silk source
In vain.Such as a kind of Publication No. CN 1969799A fibroin cosmetic product of disclosure of the invention, it is furnished with silkworm body in the fibroin cosmetic product
Silk-fibroin liquid in sericterium.The preparation method of the silk-fibroin liquid is：The silk-fibroin liquid 100 taken out out of silkworm body in sericterium
Part, 900 parts of distilled water, 10 parts of sodium hydroxide are put into enamel cylinders, then adjust pH value to 10-11；Heat temperature raising, treat silk-fibroin
Untill being completely dissolved；It is 30Be'(20 DEG C to allocate hydrolyzate concentration with distilled water again) it is standby after filtering.
The spider's thread is a kind of natural animal azelon, more containing (GPGXX) n/ (GPGQQ) n, An/ (GA) n, (GGX) n etc.
Kind repeats peptide sequence；With intensity is high, elasticity is good, initial modulus is big, energy to failure is big, biodegradable, good biocompatibility,
Moisture retention is good, other synthesized high-performance fibers incomparable excellent mechanical properties and characteristic such as slim and graceful.The traditional Chinese medical science thinks, spider
Silk, nature and flavor：Cold nature.Channel tropism：Liver Channel.Major function：Hemostasis；Disappear wart.Main haematemesis；Metal-inflicted wound bleeding；Wart；Blood knurl；Hemorrhoid complicated by anal fistula.
For spider silk containing abundant amino acid, hydrolysate is nutritious, while has the function that anti-oxidant, strengthen immunity, anti-inflammation hemostasia.
The current hydrolysis process about spider silk is rarely reported.What Sheng Jiayong was delivered《The chemical composition of spider silk and structure pre-test》Middle public affairs
A kind of method for hydrolysis of spider silk has been opened, spider silk has been placed in hydrochloric acid solution, 24h is hydrolyzed at 110 DEG C.But the method for hydrolysis
Preparation condition is harsher, and amino acid yield is relatively low.
The content of the invention
In view of this, the invention provides one kind is easy to operate, energy consumption is low, easily industrialized spider silk hydrolysate and its preparation
Method, spider silk hydrolysate hydrolysis yield are high.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：
The invention provides a kind of spider silk hydrolysate, it is made using following steps：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, the water under the conditions of 80~90 DEG C
4~5h is solved, centrifugation, obtains hydrolyzate；
Hydrolyzate pH value is adjusted to 4.0~5.5, filtering, obtains spider silk hydrolysate.
Preferably, the concentration of sodium hydroxide solution is 15~25g/L.
Preferably, the concentration of sodium hydroxide solution is 20g/L.
Preferably, the rotating speed of centrifugation is 10000~20000r/min, the time is 10~20min.
Preferably, the rotating speed of centrifugation is 12000~18000r/min, and the time is 10~20min.
Preferably, the reagent of regulation hydrolyzate pH value is hydrochloric acid solution.
Preferably, the concentration of hydrochloric acid solution is 0.5~1.5mol/L.
Preferably, the concentration of hydrochloric acid solution is 1mol/L.
Preferably, the length ＜ 1mm of the spider silk after crushing.
Preferably, the aperture of filtering is 200nm.
Preferably, the number of hydrolysis is 2 times.
In embodiment provided by the invention, when the number of hydrolysis is 2 times, spider silk hydrolysate uses following steps system
：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, the water under the conditions of 80~90 DEG C
4~5h is solved, centrifugation, obtains the first hydrolyzate and precipitation；
Precipitation is mixed with sodium hydroxide solution, 4~5h is hydrolyzed under the conditions of 80~90 DEG C, obtains the second hydrolyzate；
Merge the first hydrolyzate and the second hydrolyzate, regulation pH value to 4.0~5.5, filtering, obtain spider silk hydrolysate.
Present invention also offers the preparation method of the spider silk hydrolysate, comprise the following steps：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, the water under the conditions of 80~90 DEG C
4~5h is solved, centrifugation, obtains hydrolyzate；
Hydrolyzate pH value is adjusted to 4.0~5.5, filtering, obtains spider silk hydrolysate.
Preferably, the concentration of sodium hydroxide solution is 15~25g/L.
Preferably, the concentration of sodium hydroxide solution is 20g/L.
Preferably, the rotating speed of centrifugation is 10000~20000r/min, the time is 10~20min.
Preferably, the rotating speed of centrifugation is 10000~15000r/min, and the time is 10~20min.
Preferably, the reagent of regulation hydrolyzate pH value is hydrochloric acid solution.
Preferably, the concentration of hydrochloric acid solution is 0.5~1.5mol/L.
Preferably, the concentration of hydrochloric acid solution is 1mol/L.
Preferably, the length ＜ 1mm of the spider silk after crushing.
Preferably, the aperture of filtering is 200nm.
Preferably, the number of hydrolysis is 2 times.
In embodiment provided by the invention, spider silk hydrolysate is made using following steps：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, the water under the conditions of 80~90 DEG C
4~5h is solved, centrifugation, obtains the first hydrolyzate and precipitation；
Precipitation is mixed with sodium hydroxide solution, 4~5h is hydrolyzed under the conditions of 80~90 DEG C, obtains the second hydrolyzate；
Merge the first hydrolyzate and the second hydrolyzate, regulation pH value to 4.0~5.5, filtering, obtain spider silk hydrolysate.
The invention provides a kind of spider silk hydrolysate and preparation method thereof.The spider silk hydrolysate uses following steps system
：Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, under the conditions of 80~90 DEG C hydrolyze 4~
5h, centrifugation, obtains hydrolyzate；Hydrolyzate pH value is adjusted to 4.0~5.5, filtering, obtains spider silk hydrolysate.The present invention has
Technique effect be：
(1) preparation technology small investment of the present invention, device requirement are few；Easy to operate, energy consumption is small, and safe operation coefficient is higher；
Reagent is conventional commercial reagent, suitable enlarged experiment and industrialization.
(2) preparation technology acid hydrolysis yield of the present invention is 74.15%~80.89%, yield ＞ 70%；Hydrolyzate
In, in addition to methionine, phenylalanine, tyrosine, the equal ＞ 60% of yield of other amino acid.Amino acids distribution i.e. in hydrolyzate
It is consistent substantially with the amino acid in spider silk.
(3) hydrolyzate middle-molecular-weihydroxyethyl illustrates spider in 1000-5000D micromolecule polypeptide accounting 72.18%~76.69%
Spider's thread hydrolyzate had both maintained original beauty, medical treatment and other effects, while it is easily absorbed by the body.
Embodiment
The invention discloses a kind of spider silk hydrolysate and preparation method thereof, those skilled in the art can be used for reference in this paper
Hold, be suitably modified technological parameter realization.In particular, all similar replacements and change are to those skilled in the art
For be it will be apparent that they are considered as being included in the present invention.The method of the present invention and application are by preferably implementing
Example is described, related personnel substantially can not depart from present invention, in spirit and scope to method described herein and
Using being modified or suitably change with combining, to realize and using the technology of the present invention.
Raw materials used or reagent can be bought by market in spider silk hydrolysate provided by the invention and preparation method thereof.
Spider silk picks up from Hainan Zhu Wang pharmaceutcal corporation, Ltds the five fingers Herba Calanthes japonicae cultivation base Chilobrachys Jingzhao in following examples
Spider produces spider silk.
With reference to embodiment, the present invention is expanded on further：
The preparation of the spider silk hydrolyzate of embodiment 1
The hydrolysis process of the present embodiment spider silk hydrolyzate is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 20g/L sodium hydroxide solution 200mL,
80 DEG C of temperature, hydrolyze 5h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 4.5 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
The preparation of the spider silk hydrolyzate of embodiment 2
The hydrolysis process of the present embodiment spider silk hydrolyzate is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 15g/L sodium hydroxide solution 200mL,
85 DEG C of temperature, hydrolyze 4.5h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 5.0 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
The preparation of the spider silk hydrolyzate of embodiment 3
The hydrolysis process of the present embodiment spider silk hydrolyzate is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 25g/L sodium hydroxide solution 200mL,
90 DEG C of temperature, hydrolyze 4h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 5.5 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 1
Prepared with reference to the preparation method disclosed in Publication No. CN 1969799A patent of invention, be specially：
(1) pre-process：After taking spider silk, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider silk
Into length ＜ 1mm cotton-shaped or powdery.
(2) pretreated 100 parts of spider silk, 900 parts of distilled water are taken, 10 parts of sodium hydroxide is put into enamel cylinders；Heating
Heating, temperature are 90-100 DEG C, hydrolyze 24h；It is 30Be'(20 DEG C to allocate hydrolyzate concentration with distilled water again).
(3) filter：By hydrolyzate through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
Comparative example 2
Delivered with reference to Sheng man a large bell《The chemical composition of spider silk and structure pre-test》Disclosed in spider silk method for hydrolysis,
Specially：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) spider silk is placed in 6mol/L hydrochloric acid solution, 24h is hydrolyzed at 110 DEG C, removed in spider silk hydrolyzate
Hydrochloric acid.
Comparative example 3
Compared with Example 1, this comparative example reduces the step of hydrolysis number of times, and hydrolysis process is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 20g/L sodium hydroxide solution 200mL,
90 DEG C of temperature, hydrolyze 4h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) neutralize：PH is neutralized to 4.5 with 1mol/L hydrochloric acid solutions.
(6) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(7) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 4
Compared with Example 1, this comparative example lacks the step of pretreatment crushes, and hydrolysis process is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, load 1L glass container.
(2) hydrolyze：20g/L sodium hydroxide solution 200mL are added, 90 DEG C of temperature, hydrolyze 4h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(7) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 5
Compared with Example 1, this comparative example lacks the step of heating, and hydrolysis process is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 20g/L sodium hydroxide solution 200mL,
Normal temperature, hydrolyze 5h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 4.5 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 6
Compared with Example 1, in this comparative example, sodium carbonate replaces sodium hydroxide, and hydrolysis process is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 20g/L sodium carbonate liquor 200mL, temperature
80 DEG C of degree, hydrolyze 5h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 4.5 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 7
Compared with Example 1, hydrolysis temperature is adjusted to 100 DEG C in this comparative example, and the step of heating, hydrolysis process is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) hydrolyze：Spider silk after crushing is loaded to 1L glass container, adds 20g/L sodium hydroxide solution 200mL,
100 DEG C of temperature, hydrolyze 5h.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) hydrolyze：The spider silk of unfinished hydrolysis is hydrolyzed once again, method is the same as (2).
(6) neutralize：PH is neutralized to 4.5 with 1mol/L hydrochloric acid solutions.
(7) filter：By the hydrolyzate after neutralization through 200nm membrane filtrations, colourless or lurid spider silk hydrolyzate is obtained.
(8) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Comparative example 8
Compared with Example 1, in this comparative example, hexafluoroisopropanol dissolving replaces sodium hydroxide hydrolysis, and concrete technology is：
(1) pre-process：After taking 2g spider silks, purifying water wash to fall impurity, agate ball mill pulverization process is placed in, to spider
Cotton-shaped or powdery of the silk into length ＜ 1mm.
(2) dissolve：Spider silk after crushing is loaded to 1L glass container, adds 200mL hexafluoroisopropanol solution, room temperature
Dissolving.
(3) centrifuge：Hydrolyzate is centrifuged, centrifugal rotational speed 15000r/min, centrifuge 20min.
(4) collect：The hydrolyzate of centrifugation is collected to glass container.
(5) dissolve：The spider silk of unfinished hydrolysis is dissolved once again, method is the same as (2).
(6) filter：By the hydrolyzate after dissolving through 200nm membrane filtrations, spider silk hydrolyzate is obtained.
(7) preserve：Spider silk hydrolyzate is placed in glass container, sealed, it is stored refrigerated.
Test example 1
Reference《The measure of amino acid in GB 5009.124-2016 national food safety standard food》In detection method
Embodiment 1-3 and comparative example 1-8 amino acid content and molecular weight accounting is determined, it is as a result as follows：
Amino acid and the analysis of molecular weight accounting in the spider's thread of 1 embodiment of table 1 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 2 embodiment of table 2 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 3 embodiment of table 3 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 4 comparative example of table 1 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 5 comparative example of table 2 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 6 comparative example of table 3 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 7 comparative example of table 4 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 8 comparative example of table 5 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 9 comparative example of table 6 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 10 comparative example of table 7 and its hydrolyzate
Amino acid and the analysis of molecular weight accounting in the spider's thread of 11 comparative example of table 8 and its hydrolyzate
The total hydrolysis of the amino acid as made from above-mentioned results showed that using 1-3 of embodiment of the present invention preparation method
Yield/% is up to 74.15%~80.89%, micromolecule polypeptide accounting 72.18% of the hydrolyzate middle-molecular-weihydroxyethyl in 1000-5000D
~76.69%, it is significantly higher than comparative example 1-8 hydrolysis yield.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of spider silk hydrolysate, it is characterised in that be made using following steps：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, under the conditions of 80~90 DEG C hydrolyze 4~
5h, centrifugation, obtains hydrolyzate；
The hydrolyzate pH value is adjusted to 4.0~5.5,200nm membrane filtrations, obtains spider silk hydrolysate.
2. spider silk hydrolysate according to claim 1, it is characterised in that the concentration of the sodium hydroxide solution be 15~
25g/L。
3. spider silk hydrolysate according to claim 1, it is characterised in that the rotating speed of the centrifugation be 10000~
20000r/min, time are 10~20min.
4. spider silk hydrolysate according to claim 1, it is characterised in that the reagent of regulation hydrolyzate pH value is molten for hydrochloric acid
Liquid, the concentration of the hydrochloric acid solution is 0.5~1.5mol/L.
5. spider silk hydrolysate according to any one of claim 1 to 4, it is characterised in that the number of the hydrolysis is 2
It is secondary.
6. a kind of preparation method of spider silk hydrolysate, it is characterised in that comprise the following steps：
Spider silk is crushed, the spider silk after crushing mixes with sodium hydroxide solution, under the conditions of 80~90 DEG C hydrolyze 4~
5h, centrifugation, obtains hydrolyzate；
The hydrolyzate pH value is adjusted to 4.0~5.5,200nm membrane filtrations, obtains spider silk hydrolysate.
7. preparation method according to claim 6, it is characterised in that the concentration of the sodium hydroxide solution is 15~25g/
L。
8. preparation method according to claim 6, it is characterised in that the rotating speed of the centrifugation is 10000~20000r/
Min, time are 10~20min.
9. preparation method according to claim 6, it is characterised in that the reagent of regulation hydrolyzate pH value is hydrochloric acid solution,
The concentration of the hydrochloric acid solution is 0.5~1.5mol/L.
10. the preparation method according to any one of claim 6 to 9, it is characterised in that the number of the hydrolysis is 2 times.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101628126A (en) * | 2009-08-18 | 2010-01-20 | 四川大学 | Natural polypeptides material for biomineralization and preparation method thereof |
| WO2013152265A1 (en) * | 2012-04-06 | 2013-10-10 | Trustees Of Tufts College | Methods of producing and using silk microfibers |
| CN103739691A (en) * | 2013-12-11 | 2014-04-23 | 浙江大学 | Green and environment-friendly method for preparing silk fibroin powder |
-
2017
- 2017-11-08 CN CN201711090220.8A patent/CN107652354B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101628126A (en) * | 2009-08-18 | 2010-01-20 | 四川大学 | Natural polypeptides material for biomineralization and preparation method thereof |
| WO2013152265A1 (en) * | 2012-04-06 | 2013-10-10 | Trustees Of Tufts College | Methods of producing and using silk microfibers |
| CN103739691A (en) * | 2013-12-11 | 2014-04-23 | 浙江大学 | Green and environment-friendly method for preparing silk fibroin powder |
Non-Patent Citations (1)
| Title |
|---|
| FRITZ VOLLRATH: "Biology of spider silk", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
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