CN105669832B - A kind of polypeptide preparing hydrogel and its hydrogel of preparation - Google Patents
A kind of polypeptide preparing hydrogel and its hydrogel of preparation Download PDFInfo
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- 239000000017 hydrogel Substances 0.000 title claims abstract description 45
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 29
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 25
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title abstract description 12
- 238000006731 degradation reaction Methods 0.000 claims abstract description 11
- 230000015556 catabolic process Effects 0.000 claims abstract description 9
- 238000012258 culturing Methods 0.000 claims abstract description 3
- 239000011159 matrix material Substances 0.000 claims description 14
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 7
- 239000007853 buffer solution Substances 0.000 claims description 7
- 239000000243 solution Substances 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 claims description 5
- 230000000593 degrading effect Effects 0.000 claims description 3
- 230000021736 acetylation Effects 0.000 claims description 2
- 238000006640 acetylation reaction Methods 0.000 claims description 2
- 150000001413 amino acids Chemical group 0.000 claims description 2
- 210000004899 c-terminal region Anatomy 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 238000011534 incubation Methods 0.000 claims description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims 2
- 229910021529 ammonia Inorganic materials 0.000 claims 1
- 238000010438 heat treatment Methods 0.000 claims 1
- 239000000499 gel Substances 0.000 abstract description 17
- 102000004190 Enzymes Human genes 0.000 abstract description 5
- 108090000790 Enzymes Proteins 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 239000013043 chemical agent Substances 0.000 abstract description 4
- 208000027418 Wounds and injury Diseases 0.000 abstract description 2
- 230000006378 damage Effects 0.000 abstract description 2
- 231100000734 genotoxic potential Toxicity 0.000 abstract description 2
- 230000036541 health Effects 0.000 abstract description 2
- 230000005847 immunogenicity Effects 0.000 abstract description 2
- 208000014674 injury Diseases 0.000 abstract description 2
- 230000004962 physiological condition Effects 0.000 abstract description 2
- 125000003275 alpha amino acid group Chemical group 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 14
- 239000000523 sample Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 9
- 238000003860 storage Methods 0.000 description 8
- 229910052757 nitrogen Inorganic materials 0.000 description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000001338 self-assembly Methods 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 239000003643 water by type Substances 0.000 description 3
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical compound CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 2
- 101710123874 Protein-glutamine gamma-glutamyltransferase Proteins 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002121 nanofiber Substances 0.000 description 2
- 238000000518 rheometry Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241001474374 Blennius Species 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 1
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 108060008539 Transglutaminase Proteins 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000000816 matrix-assisted laser desorption--ionisation Methods 0.000 description 1
- 229910021421 monocrystalline silicon Inorganic materials 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 102000003601 transglutaminase Human genes 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0068—General culture methods using substrates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention provides a kind of polypeptide for preparing hydrogel, and amino acid sequence is SEQ ID NO:1；Polypeptide of the invention is used to prepare hydrogel；The hydrogel of preparation can be used for preparing cell culturing bracket.Polypeptide of the invention in physiological conditions, is catalyzed using the enzyme of organism itself, is formed hydrogel, is avoided the injury to tissue such as additional chemical agent, ultraviolet light.And degrade using biological enzyme, the degradation by organism endogenous material to gel is realized, genotoxic potential and immunogenicity risk caused by introducing external source chemical agent are avoided；It is a kind of ideal tissue engineering material, is of great significance to human life and health.
Description
Technical field
The preparation technical field of hydrogel of the present invention, and in particular to a kind of polypeptide for preparing hydrogel and its preparation
Hydrogel.
Background technique
Prior art preparation hydrogel can be divided into two class of macromolecules cross-linking and small molecule self assembly.Small molecule synthesis is more held
Easily, but due to its structure be engineer, formed gel utilize is secondary key active force, it is tired to cause gel degradation
The gel strength of difficulty, formation is limited.High-molecular gel, which has, prepares the considerable feature of simple, easy control of structure, intensity, but is difficult to reality
Existing biodegrade；It is difficult to realize the regulation to cell specific function simultaneously.Currently used high molecular material such as chitin, seaweed
Sour sodium, collagen or synthesis macromolecule such as polyethylene glycol through chemical modification etc., this kind of high molecular material has good branch
Support property, but the problems such as chemical residual in synthesis, radical initiator, ultraviolet light, limits its extensive use.
Summary of the invention
The object of the present invention is to provide a kind of polypeptide for preparing hydrogel and its hydrogels of preparation, to make up existing skill
The deficiency of art.
Present invention firstly provides a kind of polypeptide for preparing hydrogel, amino acid sequence be IIISLKGQ (SEQ ID NO:
1)；
The N-terminal of aforementioned polypeptides has carried out acetylation, and C-terminal carries out amination；
Above-mentioned polypeptide is used to prepare hydrogel；
The present invention also provides a kind of hydrogel, preparation method is as follows: being Ac-I by structural formula3SLKGQ-NH2Polypeptide exist
Heated in the buffer solution that pH is 7~8 and cooling, be then added glutamine transaminage, 37 DEG C be incubated for 24 hours after form water
Gel.
The buffer solution, preferably Hepes solution；
It is 0.1U/mL to 20U/mL, preferably 0.9U/mL that concentration range, which is added, in the glutamine transaminage；
The addition concentration of small peptide is 4-32mM, preferably 8mM；
A method of above-mentioned hydrogel of degrading is that matrix metalloproteinase-II is added；
Matrix metalloproteinase-II the concentration used is 25ng/mL to 500ng/mL, preferred concentration 100ng/mL.
The hydrogel of above-mentioned preparation is preparing the application in cell culturing bracket.
Polypeptide of the invention in physiological conditions, is catalyzed using the enzyme of organism itself, is formed hydrogel, is avoided
The injury to tissue such as additional chemical agent, ultraviolet light.And degrade using biological enzyme, it realizes through organism endogenous
Degradation of the substance to gel avoids genotoxic potential and immunogenicity risk caused by introducing external source chemical agent；Be it is a kind of more
Ideal tissue engineering material, is of great significance to human life and health.
Detailed description of the invention
Fig. 1 is Ac-I3SLKGQ-NH2Molecule is substance assistant laser desorpted ionized after matrix metalloproteinase-II degradation
Parting time mass spectrum figure；
Fig. 2 is Ac-I of the present invention3SLKGQ-NH2The atomic force of formed hydrogel after glutamine transaminage is added in molecule
Microscope shape appearance figure；
Fig. 3 is Ac-I of the present invention3SLKGQ-NH2The atomic force microscope shape appearance figure before glutamine transaminage is added in molecule；
Fig. 4 is Ac-I of the present invention3SLKGQ-NH2The mechanical strength of hydrogel before molecule is degraded through matrix metalloproteinase-II
Relationship between (storage modulus G ' and Loss modulus G ") and stress；
Fig. 5 is Ac-I of the present invention3SLKGQ-NH2Machine of the hydrogel that molecule is formed after matrix metalloproteinase-II degradation
Relationship between tool intensity (storage modulus G ' and Loss modulus G ") and stress；
Fig. 6 is invention Ac-I3SLKGQ-NH2The hydrogel that molecule is formed is degraded through matrix metalloproteinase-II
Afm scan figure afterwards.
Specific embodiment
The present invention is described in further details combined with specific embodiments below.
Specification, the model of the major experimental instrument specifically selected in the embodiment of the present invention are described briefly first, it is following
Laboratory apparatus can be bought by commercial channel and be obtained:
Microwave-assisted Peptide synthesizer (CEM company, Liberty1 type),
(2695 type separative unit of Waters company is equipped with the inspection of Waters2996 type diode array to high performance liquid chromatograph
Survey device),
Low-temperature transmission electron microscope (Cyro-TEM, JEOL1400Plus type, Japan Electronics Corporation, Japan),
Atomic force microscope (AFM, Multimode VIII type, Brooker company, Germany),
Rotational rheometer (Haake Mars type III, power & light company, the U.S.),
Desk centrifuge (Ai Bendefu company, Germany),
Carbon dioxide cell incubator (Heracell 150i type, power & light company, the U.S.),
Superclean bench (Airtech type, the safe and sound company in Jiangsu),
Grade inverted microscope (TS100 type, Nikon Corporation, Japan) is cultivated,
Fluorescence inverted microscope (DMI3000B type, Lycra company, Germany),
Disposable Tissue Culture Flask (25cm2Costar type, Corning Incorporated, the U.S.),
Disposable pipette (5mL costar type, Corning Incorporated, the U.S.),
Disposable tissue culture plate (3599 types, Corning Incorporated, the U.S.),
Disposable tissue culture plate (3548 types, Corning Incorporated, the U.S.),
Liquid nitrogen container (YDS-30-125 type, East Asia liquid nitrogen container company).
Detect Ac-I3SLKGQ-NH2Self assembly Shape measure (AFM, Cyro-TEM) side in Tris-HCl buffer
Method, the specific detection method is as follows:
AFM scan: the polypeptide sample for taking 2 μ L to prepare is added dropwise on clean monocrystalline silicon sheet surface, and it is super that 300 μ L are added dropwise rapidly
Pure water adsorbs 10s, and then high pure nitrogen dries up sample, is swept under AFM microscope with tapping-mode (tapping mode) completion
It retouches, scanning angle is 0 °, 1~1.5Hz of sweep speed, and probe is TESP-V2 type silicon probe (Brooker company, Germany), needle point
Radius is about 10nm, is raised one's arm 127 μm, coefficient of elasticity 42N/m long, and same sample scans 5 times in different location, the results show that
Ac-I3SLKGQ-NH2、Ac-I3SLKGK-NH2Nanofibrous structures, such as Fig. 2 are self-assembly of in Tris-HCl buffer
It is shown.
Cyro-TEM: 10 μ L polypeptide solutions are drawn and are added dropwise on micro-grid surface, surface liquid is sucked after adsorbing 6s, submerges rapidly
Enter in liquid ethane, be stored in liquid nitrogen after freezing 5min, uses transmission electron microscope observation later.The results show that passing through
Transmission electron microscope observation is presented as fibre structure in Tris-HCl buffer solution to two polypeptide samples, with atomic force
Microscope observes consistent, as shown in Figure 3.
Embodiment 1:
The preparation method of the present embodiment hydrogel, comprising the following steps:
By polypeptide A c-I3SLKGQ-NH2Under the action of buffer solution Hepes buffer solution, being diluted to concentration is 8mM, warp
It is vortexed, after ultrasound, is placed in 85 DEG C of water-baths and heats and be cooled to room temperature, glutamine transaminage is added
(Transglutaminase, TGase) makes the final concentration of 0.9U/mL of TGase, respectively 37 DEG C of incubations.24 hours and 15 days
Afterwards, it is capable of forming hydrogel.It is tested by rheology, as shown in figure 4, Ac-I3SLKGQ-NH2The storage modulus of polypeptide hydrogel
G ' is about 800Pa, and Loss modulus G " is about the 1/10 of storage modulus G ', illustrates System forming typical case hydrogel.
Hydrogel degradation test: at 37 DEG C, matrix metalloproteinase II is added into the hydrogel being self-assembly of
(MMP-2), being allowed to final concentration of is 100ng/mL, and gel is placed in 37 DEG C of water-baths to rheology after being incubated for 15 days and is tested, discovery
The storage modulus G ' of gel is reduced to 1/2 or so before addition matrix metalloproteinase-II, as shown in Figure 5.Illustrate water-setting
Glue is gradually degraded by matrix metalloproteinase-II.Gel is after degrading with Matrix-assisted laser desorption ionization
Characterization has the generation of expected molecule fragment as the result is shown, it was demonstrated that the generation of degradation process, as shown in Figure 1.To solidifying after degradation
Glue carries out afm scan, the results show that fiber is reduced in gel, it was demonstrated that the generation of degradation process, such as Fig. 6 institute
Show.
Embodiment 2:
Ac-I3SLKGQ-NH2The preparation method of gel:
Choose the small peptide Ac-I of aforementioned solid-phase synthesis preparation3SLKGQ-NH2Under the action of Hepes buffer, it is diluted to
Concentration is 8mM, after concussion ultrasound, is placed in 85 DEG C of water-baths and heats 2 hours, cooled to room temperature, it is slow that addition is dissolved in Hepes
In fliud flushing and contain 5mM CaCl2With the glutamine transaminage of 2mM dithiothreitol (DTT) (DTT), make the final activity of enzyme
0.9U/mL, small peptide ultimate density are 7.27mM.Acquired solution is placed in 37 DEG C of water-baths and is incubated for 24 hours, it is available to fall
The hydrogel of vertical self-supporting.
Gel in embodiment 2 is measured:
Draw 20 μ LAc-I3SLKGQ-NH2Gel is added dropwise on clean silicon wafer, is rapidly added 300 μ L ultrapure waters later,
After standing adsorption 10s, it is dried with nitrogen sample, AFM scan, after discovery forms gel, the structure of nanofiber is remained unchanged, and is such as schemed
Shown in 2.It draws 10 μ L polypeptide solutions to be added dropwise on micro-grid surface, sucks surface liquid after adsorbing 6s, be submerged into liquid ethane rapidly
In, it is stored in liquid nitrogen after freezing 5min, uses transmission electron microscope observation later.The results show that aobvious by transmitted electron
Micro mirror observes that polypeptide sample is fibre structure in Hepes buffer solution, and consistent, such as figure is arrived with atomic force microscope observation
Shown in 3.
In Ac-I3SLKGQ-NH2After gel-forming 24 hours, NIH-3T3 cell is added in side on the hydrogel, at 37 DEG C,
After cultivating 15 days in 5%CO2 environment, micro- sem observation is shown, NIH-3T3 cellular portions are the research and development of artificial tissue into inside
Provide experimental considerations.
Embodiment 3:
Ac-I is characterized using Haake torque rheometer3SLKGQ-NH2The mechanical performance (viscoplasticity) of hydrogel sample, using diameter
The cone-plate for being 2 ° for 35mm taper is rotor, uses the plate of 35mm diameter for stator, and measurement is 400 using sample volume every time
μ L, constant experimental temperature is 25 DEG C, carries out stress scans with 1Hz frequency, and scanning range is 0.01% to 100%, measures gel
Linear viscoelastic region, suitable stress is chosen from linear viscoelastic region and carries out dynamic frequency scanning, scanning range be 0.01Hz extremely
100Hz studies the relationship between storage modulus G ' and Loss modulus G ".
By Ac-I3SLKGQ-NH2Being diluted to concentration under the effect of Hepes buffer is 8mM, is added according to the method for embodiment 2
Enter glutamine transaminage, be statically placed in 37 DEG C of water-baths 24 hours, then take 400 μ L carried out under 1Hz frequency effect 0.01% to
100% stress scans, experimental result is as shown in figure 4, the storage modulus G ' of hydrogel is about 800Pa.
Using method in this example, measurement be added the matrix metalloproteinase-II of 100ng/mL and 37 DEG C be incubated for 15 days after
Hydrogel, its storage modulus G ' is about 400Pa or so to stress scans as the result is shown, is reduced to addition matrix metalloproteinase-II
Preceding half.
Embodiment 4:
Ac-I is prepared according to the method in embodiment 23SLKGQ-NH2Hydrogel, and 100ng/mL final concentration is added thereto
Matrix metalloproteinase-II, after 37 DEG C are incubated for 15 days, draw 20 μ LAc-I3SLKGQ-NH2Gel is added dropwise in clean silicon wafer
On, it is rapidly added 300 μ L ultrapure waters later, after standing adsorption 10s, is dried with nitrogen sample, AFM scan, discovery is by matrix gold
Proteases-II degradation after, nanofiber it is destructurized, as shown in Figure 6.
It should be noted that under the introduction of this specification, any equivalent way that those skilled in the art are made, or
Obvious variant should all be within the scope of the present invention.
Claims (10)
1. a kind of polypeptide, which is characterized in that its amino acid sequence of the polypeptide is SEQ ID NO:1.
2. polypeptide as described in claim 1, which is characterized in that the N-terminal of the polypeptide has carried out acetylation, and C-terminal carries out ammonia
Base.
3. polypeptide of any of claims 1 or 2 is preparing the application in hydrogel.
4. a kind of hydrogel, which is characterized in that the hydrogel be by polypeptide as claimed in claim 2 pH be 7~8 it is slow
Heating and cooling in solution are rushed, glutamine transaminage is then added, forms hydrogel after 37 DEG C of incubations.
5. hydrogel as claimed in claim 4, which is characterized in that the buffer solution is Hepes solution.
6. hydrogel as claimed in claim 4, which is characterized in that the glutamine transaminage be added concentration be 0.1~
20U/mL。
7. hydrogel as claimed in claim 4, which is characterized in that the addition concentration of the polypeptide is 4-32mM.
8. hydrogel as claimed in claim 4 is preparing the application in cell culturing bracket.
9. a kind of method for hydrogel as claimed in claim 4 of degrading, which is characterized in that be with matrix metalloproteinase-II come
Degradation.
10. method as claimed in claim 9, which is characterized in that the use concentration of the matrix metalloproteinase-II is 25
~500ng/mL.
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| CN116284341B (en) * | 2023-02-20 | 2024-05-03 | 中国石油大学(华东) | Preparation and application of deep sea fish skin collagen peptide with low immunogenicity, blood pressure reduction and oxidation resistance |
| CN117511851B (en) * | 2024-01-03 | 2024-03-26 | 中国肉类食品综合研究中心 | Application of hydrogel scaffold in preparation of cell culture meat |
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