CN108244657A - A kind of method for hydrolysis of spider silk - Google Patents
A kind of method for hydrolysis of spider silk Download PDFInfo
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- CN108244657A CN108244657A CN201810029726.6A CN201810029726A CN108244657A CN 108244657 A CN108244657 A CN 108244657A CN 201810029726 A CN201810029726 A CN 201810029726A CN 108244657 A CN108244657 A CN 108244657A
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- spider silk
- hydrolysis
- water
- sulfuric acid
- hydrolyzate
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- 229920001872 Spider silk Polymers 0.000 title claims abstract description 79
- 238000000034 method Methods 0.000 title claims abstract description 36
- 230000007062 hydrolysis Effects 0.000 title claims abstract description 31
- 238000006460 hydrolysis reaction Methods 0.000 title claims abstract description 31
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 75
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 16
- 239000003480 eluent Substances 0.000 claims abstract description 13
- 238000003756 stirring Methods 0.000 claims abstract description 13
- 239000008236 heating water Substances 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 4
- OSGAYBCDTDRGGQ-UHFFFAOYSA-L calcium sulfate Chemical compound [Ca+2].[O-]S([O-])(=O)=O OSGAYBCDTDRGGQ-UHFFFAOYSA-L 0.000 claims description 3
- 238000002390 rotary evaporation Methods 0.000 claims description 2
- 230000008719 thickening Effects 0.000 claims description 2
- 238000005903 acid hydrolysis reaction Methods 0.000 abstract description 9
- 239000002537 cosmetic Substances 0.000 abstract description 9
- 238000002156 mixing Methods 0.000 abstract description 6
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 abstract description 4
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 abstract description 3
- 235000004279 alanine Nutrition 0.000 abstract description 3
- 235000015872 dietary supplement Nutrition 0.000 abstract description 3
- 235000013402 health food Nutrition 0.000 abstract description 3
- 238000001556 precipitation Methods 0.000 abstract description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 238000012545 processing Methods 0.000 abstract description 2
- 238000012423 maintenance Methods 0.000 abstract 1
- 235000001014 amino acid Nutrition 0.000 description 30
- 150000001413 amino acids Chemical class 0.000 description 29
- 230000000052 comparative effect Effects 0.000 description 14
- 238000012360 testing method Methods 0.000 description 10
- 235000011149 sulphuric acid Nutrition 0.000 description 8
- 239000001117 sulphuric acid Substances 0.000 description 8
- 241000255789 Bombyx mori Species 0.000 description 5
- 238000004088 simulation Methods 0.000 description 5
- 210000002784 stomach Anatomy 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 238000005265 energy consumption Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 108010022355 Fibroins Proteins 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 210000000936 intestine Anatomy 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 230000009514 concussion Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000382353 Pupa Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 208000000260 Warts 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
- 238000013459 approach Methods 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- -1 compound amino acid Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- CBOIHMRHGLHBPB-UHFFFAOYSA-N hydroxymethyl Chemical compound O[CH2] CBOIHMRHGLHBPB-UHFFFAOYSA-N 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
- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Cosmetics (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to the processing technique field of food, cosmetics, more particularly to a kind of method for hydrolysis of spider silk.This method comprises the following steps：Spider silk is soaked in the sulfuric acid solution that mass percentage concentration is 80%~90%, 20~40min is stirred, is slowly added to water mixing, 100~180min of heating water bath under the conditions of 80~90 DEG C；After water-bath, calcium carbonate is added in, it is stirring while adding, until pH value is 5~7；After hydrolyzate is cooled to room temperature, water repeatedly washs precipitation, collects eluent；Eluent is concentrated, filters to get spider silk hydrolyzate.The acid hydrolysis yield of spider silk hydrolyzate made from preparation method of the present invention is high, and alanine, serine equal size are higher, and health food or cosmetics nutritional supplements can be used as to use, improve spider silk skin maintenance and health-care efficacy.
Description
Technical field
The present invention relates to the processing technique field of food, cosmetics, more particularly to a kind of method for hydrolysis of spider silk.
Background technology
Spider silk, cold nature, return liver warp are stopped blooding, disappear wart, have the work(such as anti-oxidant, strengthen immunity, anti-inflammation hemostasia
Effect.The main chemical compositions of spider silk are alanine (NH2-CH[CH3]-COOH), serine (NH2-CH[CH2OH]-COOH) and
Glycine (NH2-CH2- COOH), in addition other amino acid monomer Harmonic Protein Molecular Chains are formed.Spider silk is commonly called as " biology is just ", by force
Degree is big, and toughness is strong, and has preferable biocompatibility, and purposes is very extensive, such as available for preparing spider silk artificial skin, flesh
The new medicals materials such as tendon, cardiac stent, blood vessel, ultra-fine operation suture thread and flak jackets, the parachute of military domain etc..
Spider silk application prospect is extensive, but its degradation technique is more difficult, and conventional enzyme or diluted acid, diluted alkaline are difficult to be degraded.
Prior art sulphuric acid hydrolysis silkworm chrysalis, Luo Guilun delivered document in 05 phase in 2003《Silkworm chrysalis sulphuric acid hydrolysis prepares compound amino acid
Research》, it is that (silkworm chrysalis is 1 with sulfuric acid ratio to dilute sulphuric acid:3) heating (hyperbaric heating is more than 100 degrees Celsius) hydrolysis silkworm chrysalis
Obtain pupa albumen hydrolysate.The paper that Cao Xinzhi was delivered in 26 phases in 2013《Sulphuric acid hydrolysis fibroin prepares silk peptide
Research》, this method is using 30% sulphuric acid hydrolysis, solid-to-liquid ratio 1:40.In former approach sulphuric acid hydrolysis silkworm chrysalis method, although sulphur
Sour amount ratio is relatively low, but needs high-temperature and high-pressure conditions, very high to equipment requirement；Latter sulphuric acid hydrolysis silk, with spider silk ratio
It is closer to, but the sulfuric acid consumed is more, silk：Sulfuric acid needs the neutralization material used more, neutralizes pH than being up to 12 (40x30%)
Value is not easy to control, and this method is difficult to hydrolyze spider silk.So far there has been no spider silks easy to operate, that low energy consumption, yield is higher
The prior art of degradation.
Invention content
In view of this, the present invention provides a kind of method for hydrolysis of spider silk.The degradable spider silk of the method for hydrolysis, yield
Higher (in terms of total amino acid content), and maintain spider silk inherent advantages characteristic.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of method for hydrolysis of spider silk, include the following steps：
Step 1：Spider silk is soaked in the sulfuric acid solution that mass percentage concentration is 80%~90%, stirs 20~40min,
Water mixing is slowly added to, in 80~90 DEG C of heating water baths, hydrolyzes 100~180min；
Step 2：After hydrolysis, calcium carbonate is added in, it is stirring while adding, until pH value is 5~7；
Step 3：After hydrolyzate is cooled to room temperature, progress is precipitated to adding in the calcium sulfate salt formed after calcium carbonate using water
Eluent is collected in elution；
Step 4：Eluent is concentrated, is filtered.
Spider silk is soaked in the sulfuric acid solution that mass percentage concentration is 80%~90% by the present invention, stirs 20~40min,
Water mixing is slowly added to, 100~180min of heating water bath under the conditions of 80~90 DEG C；After water-bath, calcium carbonate, Bian Jia are added in
Side is stirred, until pH value is 5~7；After hydrolyzate is cooled to room temperature, water repeatedly washs the precipitation of calcium sulfate salt, collects elution
Liquid；Eluent is concentrated, filters to get spider silk hydrolyzate.The amino of spider silk hydrolyzate made from preparation method of the present invention
Sour water solution yield is high, and alanine, serine equal size are higher, and health food or cosmetics nutritional supplements can be used as to use, carried
High spider silk beauty and health-care efficacy.
In embodiment provided by the invention, the usage ratio of spider silk and sulfuric acid solution is 1：(6.0~10.0), hydrolysis
Time for 100~180min, the temperature of hydrolysis is 80~90 DEG C, and the speed that step 1 stirs is 20~30r/min.
Preferably, the mass fraction of sulfuric acid is 85% in step 1.
Preferably, in terms of g/mL, the usage ratio of spider silk and sulfuric acid solution is 1 in step 1：8.
Preferably, the speed stirred in step 1 is 25r/min.
Preferably, the volume ratio of sulfuric acid solution and water is 1 in step 1：(0.5~2.0).
In embodiment provided by the invention, the volume ratio of sulfuric acid solution and water is 1 in step 1：1.5.
Preferably, the addition speed of calcium carbonate described in step 2 is 20~50g/min.
Preferably, in terms of g/mL, the usage ratio of spider silk and water in step 3 is 1：(200~400).
Preferably, the number eluted described in step 3 is 5~8 times.
Preferably, the room temperature in step 3 is 15~35 DEG C.
Preferably, the step of further including concentration, constant volume after being filtered in step 4, filtering again.
Preferably, concentration is concentrated for rotary evaporation.
Preferably, thickening temperature is 50~80 DEG C.
Preferably, in terms of g/mL, the ratio of eluent is 1 after spider silk and step 4 concentrate in step 1：(80~
150)。
The present invention provides a kind of method for hydrolysis of spider silk, and this method comprises the following steps：Spider silk is soaked in matter
The sulfuric acid solution that percentage concentration is 80%~90% is measured, 20~40min is stirred, adds water and mixes, the water under the conditions of 80~100 DEG C
Solve 100~180min；After hydrolysis, calcium carbonate is then added in, is stirred, until pH value is 5~7；It is dense after being cooled to room temperature
Contracting, filtering.The present invention has the following technical effect that：
1st, the amino acid yield of spider silk hydrolyzate made from preparation method of the present invention is high, wherein rich in glycine, the third ammonia
The aminoacid ingredients such as acid, serine, nutrition and cosmetic values are high.
2nd, the method for the present invention spider silk nutritive loss is small, improves the nutritive value and application value of spider silk, Ke Yizuo
It is used for health food or cosmetics nutritional supplements.
Specific embodiment
The invention discloses a kind of method for hydrolysis of spider silk, those skilled in the art can use for reference present disclosure, suitably
Modified technique parameter is realized.In particular, it should be pointed out that all similar substitutions and modifications are for a person skilled in the art
It will be apparent that they are considered as being included in the present invention.The method of the present invention and application are carried out by preferred embodiment
Description, related personnel significantly can not depart from the content of present invention, in spirit and scope to method described herein and application into
Row change is suitably changed with combining, to realize and using the technology of the present invention.
Agents useful for same or instrument can be bought by market in the method for hydrolysis of spider silk provided by the invention, and spider silk is by sea
Nan Zhu king pharmaceutcal corporation, Ltd provides.
With reference to embodiment, the present invention is further explained：
Embodiment 1
Spider silk 100g is put into 2L glass reaction kettles, adds in the 600mL concentrated sulfuric acid solutions that mass fraction is 80%, leaching
Spider silk 20min is steeped, and is stirred continuously, mixing speed 25r/min (similarly hereinafter) is slow added into 300mL water, stirring, and then 80
DEG C heating water bath, and keep the temperature 100min.After heating water bath, calcium carbonate is slowly added to, adds in speed 20g/min, side edged
Stirring.PH is detected, pH stops adding calcium carbonate in 5-7.After being cooled to room temperature, divided 5 times with 20L pure water, eluted, collection is washed
De- liquid.Eluent is concentrated to 8L with Rotary Evaporators to get spider silk hydrolyzate, as cosmetics, health products.By GB
5009.124-2016 method measures the content of amino acid in hydrolyzate, 1 the results are shown in Table：
1 spider silk hydrolyzate amino acid testing result of table
Embodiment 2
Spider silk 100g is put into 5L glass reaction kettles, adds in the 800mL concentrated sulfuric acid solutions that mass fraction is 85%, leaching
Spider silk 30min is steeped, and is stirred continuously, mixing speed 25r/min (similarly hereinafter) is slow added into 1200mL water, stirring, and then 85
DEG C heating water bath, and keep the temperature 150min.After heating water bath, calcium carbonate is slowly added to, adds in speed 20g/min, side edged
Stirring.PH is detected, pH stops adding calcium carbonate in 5-7.After being cooled to room temperature, divided 6 times with 30L pure water, eluted, collection is washed
De- liquid.Eluent is concentrated to 12L with Rotary Evaporators to get spider silk hydrolyzate, as cosmetics, health products.By GB
5009.124-2016 method measures the content of amino acid in hydrolyzate, 2 are the results are shown in Table：
2 spider silk hydrolyzate amino acid testing result of table
Embodiment 3
Spider silk 100g is put into 5L glass reaction kettles, adds in the 1000mL concentrated sulfuric acid solutions that mass fraction is 90%,
Spider silk 40min is impregnated, and is stirred continuously, mixing speed 25r/min (similarly hereinafter) is slow added into 2000mL water, stirring, then
90 DEG C of heating water baths, and keep the temperature 180min.After heating water bath, calcium carbonate is slowly added to, adds in speed 20g/min, Bian Jia
It stirs on side.PH is detected, pH stops adding calcium carbonate in 5-7.After being cooled to room temperature, divided 8 times with 40L pure water, eluted, collected
Eluent.Eluent is concentrated to 15L with Rotary Evaporators to get spider silk hydrolyzate, is used as cosmetics, health products.It presses
GB 5009.124-2016 methods measure the content of amino acid in hydrolyzate, the results are shown in Table 3：
3 spider silk hydrolyzate amino acid testing result of table
Comparative example 1
The mass fraction of sulfuric acid is adjusted to 75%, other parameter the results are shown in Table 4 with embodiment 2.
Comparative example 2
The mass fraction of sulfuric acid is adjusted to 95%, other parameter the results are shown in Table 4 with embodiment 2.
4 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 4, for sulfuric acid mass fraction less than 80% or higher than 90%, amino acid yield is below embodiment
1。
Comparative example 3
The time that sulfuric acid is impregnated to spider silk is adjusted to 15min, and other parameter the results are shown in Table 5 with embodiment 2.
Comparative example 4
The time that sulfuric acid is impregnated to spider silk is adjusted to 50min, and other parameter the results are shown in Table 5 with embodiment 2.
5 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 5, the time that sulfuric acid impregnates spider silk is less than 20min, and amino acid yield can conspicuousness reduction.Together
When, the time that sulfuric acid impregnates spider silk is more than 40min, not only influences production efficiency, but also amino acid yield decreases instead.
Comparative example 5
The dosage for impregnating spider silk sulfuric acid is adjusted to 500mL, other parameter the results are shown in Table 6 with embodiment 2.
Comparative example 6
The dosage for impregnating spider silk sulfuric acid is adjusted to 1100mL, other parameter the results are shown in Table 6 with embodiment 2.
6 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 6, the sulfuric acid solution dosage and spider silk ratio are less than 6:1, due to acid insufficient, spider silk
Degradation is not thorough, and amino acid yield conspicuousness reduces.Meanwhile the sulfuric acid solution dosage and spider silk ratio are more than 10:1, instead
And causing production operation cumbersome due to follow-up calcium carbonate dosage is larger, the more, concentration time of precipitation extends etc., amino acid damage
It loses and energy consumption improves.
Comparative example 7
The water bath heating temperature for impregnating spider silk sulfuric acid is adjusted to 75 DEG C, other parameter the results are shown in Table with embodiment 2
7。
Comparative example 8
The water bath heating temperature for impregnating spider silk sulfuric acid is adjusted to 95 DEG C, other parameter the results are shown in Table with embodiment 2
7。
7 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 7, for bath temperature less than 80 DEG C or higher than 90 DEG C, acid hydrolysis yield is below embodiment
1。
Comparative example 9
The heating water bath time for impregnating spider silk sulfuric acid is adjusted to 90min, as a result other parameter is shown in embodiment 2
Table 8.
Comparative example 10
The heating water bath time for impregnating spider silk sulfuric acid is adjusted to 190min, as a result other parameter is shown in embodiment 2
Table 8.
8 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 8, water bath time is less than 100min, due to acidolysis time deficiency, leads to hydrolysis amino acid yield
It is relatively low.Meanwhile hydrolysis time, higher than 180min, amino acid yield does not improve, and production process increases energy consumption and reduction instead
Production efficiency.
Comparative example 11
Washing steps are adjusted to 4 times, other parameter the results are shown in Table 9 with embodiment 2.
Comparative example 12
Washing steps are adjusted to 9 times, other parameter the results are shown in Table 9 with embodiment 2.
9 spider silk hydrolyzate amino acid testing result of table
It can be obtained by table 9, washing steps are less than 5 times, and amino acid yield is relatively low.Meanwhile washing steps are more than 8 times, ammonia
Base acid yield does not obtain the raising of conspicuousness, increases time and the energy consumption of later stage concentration instead.
Comparative example 13
It is delivered according to Cao Xinzhi etc.《Sulphuric acid hydrolysis fibroin prepares the research of fibroin skin》, prepare spider silk hydrolysis
Liquid detects amino acid content, the results are shown in Table 10.
10 spider silk hydrolyzate amino acid testing result of table
It is 80%~90% to work as sulfuric acid concentration it can be seen from the above results, 20~40min of sulfuric acid solution, bath temperature
In 80~90 DEG C, water bath time in 100~180min, elution 5~8 times, spider silk when pure water ratio 200~400 is used in elution
Percent hydrolysis is significantly higher than the percent hydrolysis of other technological parameters.
Embodiment 4
It simulates human body stomach and absorbs environment, establish model, investigate spider silk hydrolyzate in human stomach and the absorbing state of intestines.
1st, stomach simulation digestion model
Spider silk hydrolyzate 50mL is taken, is added in 250mL conical flasks, pH=1.3~1.5 are adjusted with 1mol/L HCl,
The stomach simulated solution 2.0mL (0.2g Sigma P7000 pepsins are dissolved in 5mL0.01mol/L HCl) added, sealing, 36~
38 DEG C, 100~120min is vibrated, concussion rate obtains stomach simulation digest in 60~80r/min.0.01mol/L HCl solutions
Do blank group.
2nd, small intestine simulation digestion model
With the NaHCO of 1mol/L3Stomach simulation digest is neutralized to pH=5.0~5.5 by solution, adds in the simulation of 2.0mL intestines
(4g Sigma P7500 trypsase, 25g Sigma B8631 cholate are dissolved in 1L 0.1mon/LNaHCO to liquid3), sealing, 36~
38 DEG C, 100~120min is vibrated, concussion rate maintains pH=7.0 with 0.1moi/L, obtain intestines mould in 60~80r/min, process
Intend digest.5000r/min is centrifuged, and filtering takes supernatant, the detection of total amino acid content is carried out with amino-acid analyzer.
0.1mon/L NaHCO3Solution does blank group.
3rd, body absorption rate Q=(c0-c1)/c0, the results are shown in Table 11.
11 body absorption rate of table
In Examples 1 to 3 it can be seen from the above results, the amino acid body absorption rate of spider silk hydrolyzate is significantly high
In comparative example 13 and blank group.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method for hydrolysis of spider silk, which is characterized in that include the following steps：
Step 1：Spider silk is soaked in the sulfuric acid solution that mass percentage concentration is 80%~90%, stirs 20~40min, slowly
It adds water and mixes, in 80~90 DEG C of heating water baths, hydrolyzes 100~180min；
Step 2：After hydrolysis, calcium carbonate is added in, it is stirring while adding, until pH value is 5~7；
Step 3：After hydrolyzate is cooled to room temperature, is eluted, received to adding in the calcium sulfate salt formed after calcium carbonate using water
Collect eluent；
Step 4：Eluent is concentrated, is filtered.
2. method for hydrolysis according to claim 1, which is characterized in that in terms of g/mL, the spider silk and the sulfuric acid are molten
The usage ratio of liquid is 1：(6.0~10.0).
3. method for hydrolysis according to claim 1, which is characterized in that the speed stirred in the step 1 is 20~30r/
min。
4. method for hydrolysis according to claim 1, which is characterized in that the body of sulfuric acid solution described in step 1 and the water
Product is than being 1：(0.5~2.0).
5. method for hydrolysis according to claim 1, which is characterized in that the addition speed of calcium carbonate described in step 2 is 20
~50g/min.
6. method for hydrolysis according to claim 1, which is characterized in that in terms of g/mL, the spider silk and water in step 3
Usage ratio is 1：(200~400).
7. method for hydrolysis according to claim 1, which is characterized in that the number eluted described in step 3 is 5~8 times.
8. method for hydrolysis according to claim 1, which is characterized in that concentration, constant volume, again are further included after being filtered in step 4
The step of secondary filtering.
9. method for hydrolysis according to claim 8, which is characterized in that the concentration is concentrated for rotary evaporation, thickening temperature
It is 50~80 DEG C.
10. method for hydrolysis according to claim 1, which is characterized in that in terms of g/mL, spider silk and step 4 are dense in step 1
The ratio of eluent is 1 after contracting：(80~150).
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| CN108968059A (en) * | 2018-09-28 | 2018-12-11 | 青岛大学 | A kind of spider silk composite nutrient-fluid |
| CN108968060A (en) * | 2018-09-28 | 2018-12-11 | 青岛大学 | A kind of preparation method of spider silk composite nutrient-fluid |
| CN109329911A (en) * | 2018-09-28 | 2019-02-15 | 青岛大学 | A kind of spider silk composition oral liquid |
| CN109329910A (en) * | 2018-09-28 | 2019-02-15 | 青岛大学 | A kind of preparation method of spider silk composition oral liquid |
| CN113170898A (en) * | 2021-05-06 | 2021-07-27 | 海南蛛王生物科技有限公司 | Health-care composition assisting in reducing blood sugar and blood fat and health-care product thereof |
| CN113180248A (en) * | 2021-05-14 | 2021-07-30 | 海南蛛王生物科技有限公司 | Composition with effects of dispelling effects of alcohol and protecting liver and preparation method and health-care product thereof |
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| CN109329910A (en) * | 2018-09-28 | 2019-02-15 | 青岛大学 | A kind of preparation method of spider silk composition oral liquid |
| CN113170898A (en) * | 2021-05-06 | 2021-07-27 | 海南蛛王生物科技有限公司 | Health-care composition assisting in reducing blood sugar and blood fat and health-care product thereof |
| CN113180248A (en) * | 2021-05-14 | 2021-07-30 | 海南蛛王生物科技有限公司 | Composition with effects of dispelling effects of alcohol and protecting liver and preparation method and health-care product thereof |
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