CN110508322A - A kind of self-assembled nanometer material and the preparation method and application thereof - Google Patents
A kind of self-assembled nanometer material and the preparation method and application thereof Download PDFInfo
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Abstract
The present invention provides self-assembled nanometer material and the preparation method and application thereof, the raw material of the nano material includes nucleic acid molecules, aminoglycoside molecule and metal protoporphyrin；Self-assembled nanometer material provided by the present invention, use aminoglycoside molecule as activator, the nano material prepared is assembled altogether with nucleic acid molecules and metal protoporphyrin, as a kind of simulated enzyme catalyst, with the substrate conversion number and catalytic efficiency with natural horseradish peroxidase same order, simultaneously compared to aminoglycoside molecule and metal protoporphyrin, nucleic acid metal protoporphyrin conjugate or nucleic acid metal protoporphyrin self-assembly, aminoglycoside molecule and nucleic acid have collaboration facilitation to the catalytic activity of metal protoporphyrin, have higher catalytic activity；The application value with higher in chemical sensor and biosensor.
Description
Technical field
The invention belongs to field of nanometer material technology, it is related to a kind of self-assembled nanometer material and the preparation method and application thereof, especially
It is related to a kind of Supramolecular self assembly analogue enztme and the preparation method and application thereof.
Background technique
Peroxidase based on four serobila of nucleic acid and iron-protoporphyrin coordination assembling has important in biosensor
Application value, and four serobila of nucleic acid and the coordination complex of copper ion also show to react Diels-Alder chiral selects
Selecting property.Will by special designing, with four serobila sequence of nucleic acid DNA probe identification target (nucleic acid molecules, biological micromolecule or
Protein molecule etc.) after, releasable or exposure four serobila of nucleic acid, and then be coordinated with iron-protoporphyrin, it is catalyzed with H2O2For oxidisability bottom
The redox reaction of object, and when reproducibility substrate is that 2,2'- joins bis- (3- ethyl benzo thiazole phenanthroline -6- sulfonic acid) di-ammonium salts of nitrogen
(ABTS2-), tetramethyl benzidine (TMB) or when 10- acetyl group -3,7- dihydroxy phenoxazine (Amplex Red), oxidation product
With color, perhaps fluorescent emission can detect target molecules by absorption spectrum or fluorescent spectrometry.Meanwhile with class peroxidating
The catalysis reaction of object DNA enzymatic is that the detection method that signal is read is not necessarily to that DNA molecular is marked, easy to operate, great convenience
The development of biosensor based on DNA identification.But the substrate conversion rate and catalytic efficiency phase of class peroxide DNA enzymatic
For natural horseradish peroxidase, the general low 2-4 order of magnitude, and generally require metal cation (such as K of high concentration+, Na+, NH4 +Deng) exist and show higher catalytic activity, limit the catalytic performance and application range of fermentoid.
Develop a variety of methods at present and improves class peroxide DNA enzymatic catalytic activity.First, the aglucon for passing through index concentration
Phyletic evolution technology (SELEX) screening and optimizing DNA molecular sequence improves the stability of DNA/ iron-protoporphyrin compound, still
SELEX technology is very time-consuming, and failure rate is higher.Second, leading to by by DNA aptamers and the chemical coupling of fermentoid DNA sequence dna
The specific recognition for crossing DNA aptamers Yu reproducibility substrate (dopamine, DOPA and N- hydroxyl-arginine), by substrate in fermentoid
DNA/ iron-protoporphyrin binding site is nearby enriched with, and improves the catalysis oxidation efficiency to substrate；But since DNA aptamers can recognize bottom
Object limited types, and the molar extinction coefficient of oxidation product is not high (relative to common ABTS or TMB etc.), limits fermentoid
Catalytic oxidation type.Third, fermentoid DNA sequence dna and polyhistidyl molecule are assembled altogether, then it is coordinated with iron-protoporphyrin
Assembling；Iron-protoporphyrin proximally and distally, the guanine base of DNA is cooperateed with the histidine in polypeptide improves iron-protoporphyrin
Catalytic rate, but substrate conversion number is still low more than natural horseradish peroxidase with catalytic efficiency, does not have chiral substrates
Selectivity, and used polypeptide by solid-phase synthesis obtain, it is expensive (9mg, 20 histidines, purity >
98%, 1623 yuan are needed, Shanghai Zi Yu Biotechnology Co., Ltd), limit the further exploration of the method.
For polypeptide, aminoglycoside molecule have it is from a wealth of sources, be easily obtained, cheap (chitosan
500g, deacetylation >=95%, viscosity 100-200mPas, 230 yuan or so；Paromomycin sulfate, 100g, 350 yuan or so),
Moreover, the pKa value of aminoglycoside molecule amino is 6.0-6.5, acid-base catalysis can be played in ferroheme catalytic oxidation
Agent effect.
Currently, aminoglycoside molecule and nucleic acid, ferroheme by design are total to group and are filled with raising Mimetic Peroxidase
Substrate conversion number and catalytic efficiency do not have any research, and this method is applied to the detection of nucleic acid recognizing target, have it is easy to operate,
The features such as at low cost, detection response speed and high sensitivity.
In conclusion peroxide Nucleic acid mimics enzyme have become analysis detection and chiral catalysis field show it is potential excellent
Gesture develops a kind of nano material that aminoglycoside molecule is compound with nucleic acid and iron-protoporphyrin, self-assembly system can be improved
The responsiveness of catalytic activity, catalytic selectivity, bio-sensing, while detection limit and operating cost are reduced, it is urged for self assembly is bionical
The development in change field provides research direction.
Summary of the invention
The purpose of the present invention is to provide a kind of self-assembled nanometer materials and the preparation method and application thereof.
In order to achieve that object of the invention, the invention adopts the following technical scheme:
In a first aspect, the raw material of the nano material includes nucleic acid point the present invention provides a kind of self-assembled nanometer material
Son, aminoglycoside molecule and metal protoporphyrin.
Self-assembled nanometer material provided by the invention uses aminoglycoside molecule as activator, with nucleic acid molecules and
Metal protoporphyrin assembles the nano material prepared altogether, as a kind of simulated enzyme catalyst, has and natural horseradish peroxidase
The substrate conversion number and catalytic efficiency of enzyme same order, while compared to aminoglycoside molecule and metal protoporphyrin, nucleic acid
Metal protoporphyrin conjugate or nucleic acid metal protoporphyrin self-assembly, aminoglycoside molecule and nucleic acid urge metal protoporphyrin
Changing activity has collaboration facilitation, has higher catalytic activity, and conversion number of the hydrogen peroxide on metal protoporphyrin surface is
0.5-50s-1, catalytic reduction efficiency 0.5-10mM-1·s-1, conversion number of the reproducibility substrate on metal protoporphyrin surface be
0.1-20s-1, catalysis oxidation rate is 0.5-50 μM-1·s-1, catalytic effect protrusion.
In the present invention, metal protoporphyrin, nucleic acid molecules and aminoglycoside molecule can directly be formed by self assembly
The nano material of self assembly；Can also obtain the compound of metal protoporphyrin and nucleic acid molecules by being covalently attached, then with amino
Glycoside molecule carries out self assembly and obtains self-assembled nanometer material.
In the present invention, when passing through covalent linkage, in the terminal modified nucleic acid molecules and carboxylic for having alkynyl functionality in the end 5' or 3'
The metal protoporphyrin that base location is modified with nitrine functional group is reacted by click chemistry to be coupled.
Preferably, the nucleic acid molecules are DNA or RNA.
In the present invention, nucleic acid molecules, which are rich in, guanine deoxyribonucleotide (dGMP) or guanosine ribonucleoside acid
(GMP)。
Preferably, in DNA, molar ratio shared by guanine deoxyribonucleotide is 10%-100%, such as can
To be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.
Preferably, in DNA, guanine deoxyribonucleotide number is 4-59, such as can be 4,8,10
A, 12,15,18,20,26,29,31,33,35,40,45,50,55 or 59.
Preferably, in RNA, molar ratio shared by guanosine ribonucleoside acid is 10%-100%, such as be can be
10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.
Preferably, in RNA, guanosine ribonucleoside acid number is 4-59, such as can be 4,8,10,12
A, 15,18,20,26,29,31,33,35,40,45,50,55 or 59.
Preferably, the nucleic acid molecules are DNA, and molar ratio shared by guanine deoxyribonucleotide is 50%-
100%, guanine deoxyribonucleotide number is 19-24.
Preferably, the aminoglycoside molecule includes chitosan, paromomycin sulfate, streptomysin, neomycin, A meter Ka
Any one in star or tobramycin.
Preferably, the aminoglycoside molecule is chitosan.
Preferably, the deacetylation of the chitosan is 10%-100%, for example, can be 10%, 20%, 30%,
40%, 50%, 60%, 70%, 80%, 90% or 100%, preferably 95%-100%.
In the present invention, the deacetylation range of choice of chitosan is larger, generally, the exposed amino having in chitosan
It each may participate in reaction.
Preferably, the viscosity of the chitosan be 100mPas-800mPas, such as can be 100mPas,
200mPas, 300mPas, 400mPas, 500mPas, 600mPas, 700mPas or 800mPas, preferably
100mPa·s-200mPa·s。
Preferably, the metal protoporphyrin includes iron-protoporphyrin, Copp, manganese protoporphyrin, nickel protoporphyrin, copper original porphin
Any one in quinoline, ruthenium protoporphyrin, rhodium protoporphyrin or palladium protoporphyrin.
Preferably, the metal protoporphyrin is iron-protoporphyrin.
Preferably, the iron-protoporphyrin is hemin or hemn, preferably hemin.
Second aspect, the present invention provides a kind of preparation methods of self-assembled nanometer material as described in relation to the first aspect, will
After nucleic acid molecules and aminoglycoside molecular mixing, metal protoporphyrin is dissolved in buffer solution carry out self assembly obtain it is described from
Assemble nanometer material.
In the present invention, when preparing self-assembled nanometer material, metal protoporphyrin need to be finally added in solution；In addition,
It can also will be coupled after nucleic acid molecules and metal protoporphyrin dosage form covalent coupling, then be carried out with aminoglycoside molecule from group
Dress obtains nano material.
In the present invention, preparation method is simple, and the synthetic technology of nucleic acid is mature, aminoglycoside molecular origin extensively, valence
Lattice are cheap, therefore self-assembled nanometer material has great advantages in synthetic operation, preparation price.
Preferably, the buffer solution includes NaH2PO4/Na2HPO4Buffer solution, KH2PO4/K2HPO4Buffer solution,
It is any in Tris/Tris hydrochloric acid salt buffer solution, HEPES/HEPES sodium salt buffer solution or MES/MES sodium salt buffer solution
It is a kind of.
Preferably, the pH value of the buffer solution is 3-10, such as can be 3,4,5,6,7,8,9 or 10 etc..
Preferably, the buffer solution is MES/MES sodium salt buffer solution；PH value is preferably 6.5-7.5.
In the present invention, it is preferred to the use of pH value be the buffer solution within the scope of 6.5-7.5, the self assembly prepared at this time is received
Catalytic activity possessed by rice material is higher, and effect is more prominent.
Preferably, the aminoglycoside molecular concentration is 0.001mg/mL-0.5mg/mL, such as can be 0.001mg/
ML, 0.01mg/mL, 0.02mg/mL, 0.1mg/mL, 0.2mg/mL, 0.3mg/mL, 0.4mg/mL or 0.5mg/mL etc., preferably
0.02mg/mL-0.1mg/mL。
Preferably, the nucleic acid molecules concentration is 0.1nM-250 μM, such as can be 0.1nM, 1nM, 10nM, 1 μM, 80 μ
M, 90 μM, 100 μM, 110 μM, 150 μM, 160 μM, 180 μM, 200 μM, 210 μM, 230 μM or 250 μM etc..
Preferably, the metal protoporphyrin concentration is 0.1nM-150 μM, such as can be 0.1nM, 1nM, 5nM, 1 μM, 10
μM, 20 μM, 50 μM, 60 μM, 80 μM, 90 μM, 100 μM, 120 μM, 130 μM, 140 μM or 150 μM etc., preferably 100nM-10.0
μM。
Preferably, the temperature of the self assembly is 4 DEG C -45 DEG C, such as can be 4 DEG C, 5 DEG C, 10 DEG C, 15 DEG C, 20 DEG C, 25
DEG C, 30 DEG C, 35 DEG C, 40 DEG C or 45 DEG C etc., preferably 20 DEG C -30 DEG C.
Preferably, the time of the self assembly be 10s-60min, such as can be 10s, 50s, 1min, 10min,
20min, 30min, 40min, 50min or 60min etc., preferably 10min-30min.
The third aspect, the present invention provides a kind of catalyst, the catalyst includes self assembly as described in relation to the first aspect
Nano material.
Preferably, the catalyst has catalyzing hydrogen peroxide activity, catalysed reductive substrate active, chiral peroxide
Substrate selective oxidation activity and chiral reduction substrate selective oxidation activity.
Preferably, the reproducibility substrate includes tetramethyl benzidine, phenol, pyrogallol, dopamine, cresols, amino
Any one in phenol, dinitro-o-cresol, naphthols, pentachlorophenol, luminol or 10- acetyl group -3,7- dihydroxy phenoxazine.
Preferably, the chiral reduction substrate include DOPA enantiomter, N- hydroxyl-arginine enantiomter,
Any one in arginine enantiomter or tyrosine derivative enantiomter.
In the present invention, every catalytic performance of catalyst is specific as follows:
Hydrogen peroxide converts number catalyst surface as 0.5-50s-1, such as can be 0.5s-1、1s-1、10s-1、15s-1、20s-1、25s-1、30s-1、35s-1、40s-1、45s-1Or 50s-1Deng.
Hydrogen peroxide is 0.5-10mM by the efficiency of catalysis reduction-1·s-1, such as can be 0.5mM-1·s-1、1mM-1·
s-1、2mM-1·s-1、3mM-1·s-1、4mM-1·s-1、5mM-1·s-1、6mM-1·s-1、7mM-1·s-1、8mM-1·s-1、9mM-1·
s-1Or 10mM-1·s-1Deng.
Reproducibility substrate converts number catalyst surface as 0.1-20s-1, such as can be 0.1s-1、2s-1、5s-1、
10s-1、12s-1、14s-1、15s-1、16s-1、17s-1、18s-1、19s-1Or 20s-1。
Reproducibility substrate is 0.5-50 μM by the rate of catalysis oxidation-1·s-1, 0.5 μM-1·s-1、1μM-1·s-1、5μM-1·s-1、10μM-1·s-1、15μM-1·s-1、20μM-1·s-1、25μM-1·s-1、30μM-1·s-1、35μM-1·s-1、40μM-1·s-1、45μM-1·s-1Or 50 μM-1·s-1。
Preferably, the enantiomeric excess value of the chiral reduction substrate is 10%-99.9%.
Fourth aspect, the present invention provides a kind of self-assembled nanometer materials as described in relation to the first aspect in chemical sensor or
Application in biosensor.
Preferably, the chemical sensor is antibiotic molecule sensor.
Preferably, the biosensor is antibody sensor, antigen sensor, Protein sensor, nucleic acid sensor
Or any one in nucleic acid recognizing substrate sensor.
The present invention can be monitored mimetic enzyme catalysis reaction by extinction photometer or fluophotometer, Ke Yizuo
Concentration and the type etc. of targeted biochemical matter are detected for biology or chemical sensor elements, it is particularly possible to detect aminoglycoside point
Son, nucleic acid molecules, aptamer concentration of substrate etc..
Compared with the existing technology, the invention has the following advantages:
Self-assembled nanometer material provided by the invention uses aminoglycoside molecule as activator, with nucleic acid molecules and
Metal protoporphyrin assembles the nano material prepared altogether, as a kind of simulated enzyme catalyst, has and natural horseradish peroxidase
The substrate conversion number and catalytic efficiency of enzyme same order, while compared to aminoglycoside molecule and metal protoporphyrin, nucleic acid
Metal protoporphyrin conjugate or nucleic acid metal protoporphyrin self-assembly, aminoglycoside molecule and nucleic acid urge metal protoporphyrin
Changing activity has collaboration facilitation, has higher catalytic activity, and conversion number of the hydrogen peroxide on metal protoporphyrin surface is
0.5-50s-1, catalytic reduction efficiency 0.5-10mM-1·s-1, conversion number of the reproducibility substrate on metal protoporphyrin surface be
0.1-20s-1, catalysis oxidation rate is 0.5-50 μM-1·s-1, catalytic effect protrusion.
Self-assembled nanometer material feedstock provided by the invention is cheap, is convenient for scale application, and the self assembly of preparation is received
Rice material has higher hydrogen peroxide catalyzed and reproducibility substrate catalytic kinetic parameter, tetramethyl is joined as analogue enztme
Aniline oxidation, substrate conversion number can reach 15-40s-1, catalytic efficiency can reach 180-350mM-1·s-1, also for hydrogen peroxide
Original, substrate conversion number are 15-45s-1, catalytic efficiency 1.2-2.5mM-1·s-1, to improve sexual valence in terms of sensor application
Than that can also play a role in the removal of water body phenolic comp ' ds pollution.
Detailed description of the invention
Fig. 1 is self-assembled nanometer material electron microscope prepared by the embodiment of the present invention 4.
Fig. 2 is that self-assembled nanometer material is catalyzed H in the present invention2O2Aoxidize tetramethyl benzidine reaction monitoring curve comparison figure.
Specific embodiment
The technical scheme of the invention is further explained by means of specific implementation.Those skilled in the art should be bright
, the described embodiments are merely helpful in understanding the present invention, should not be regarded as a specific limitation of the invention.
Embodiment 1
The present embodiment is prepared by the following method self-assembled nanometer material
To the Na of 96 μ L 50mM2HPO4/NaH2PO4The G-DNA of 1 μ L100 μM is sequentially added in buffer solution (pH 7.0),
The chlorination of chitosan molecule (viscosity 200mPas, the aqueous solution containing 1% acetic acid) and 100 μM of 1 μ L of 2 μ L 1mg/mL
Ferroheme (is dissolved in dimethyl sulfoxide), and it is 10s that time interval, which is added, in sample, 30min is then mixed at 25 DEG C, reaction obtains
Analogue enztme, i.e. self-assembled nanometer material.
Wherein, the sequence of G-DNA is GGGTAGGGCGGGCGGG, is purified by HPLC or PAGE.
Catalytic activity test
1 μ L 100mM tetramethyl benzidine (being dissolved in DMSO) and 1 μ L are sequentially added into the simulation enzyme solutions of 100 μ L
Interval time is added as 5s in 200mM aqueous hydrogen peroxide solution, sample, and recording wavelength is the trap (tetramethyl benzidine of 652nm
Intermediate oxidation product) change with time, pass through the molar extinction coefficient (39000M of tetramethyl benzidine oxidation product
cm-1), calculate H2O2Rate of reduction and tetramethyl benzidine oxidation rate.
Wherein, tetramethyl benzidine is oxidized with the rate of 80nM/s.
Embodiment 2
The present embodiment is prepared by the following method self-assembled nanometer material
To the K of 90 μ L 50mM2HPO4/KH2PO4The G-DNA of 1 μ L100 μM, 8 μ are sequentially added in buffer solution (pH 7.0)
The hemin (being dissolved in dimethyl sulfoxide) of 100 μM of the paromomycin sulfate aqueous solution of L 2mM/mL and 1 μ L, sample adds
It is divided into 10s between the angle of incidence, 30min is then mixed at 25 DEG C, reaction obtains analogue enztme, i.e. self-assembled nanometer material.
Wherein, the sequence of G-DNA is GGTAGGCGGCGGTGGCGGCGGAGG, is purified by HPLC or PAGE.
Catalytic activity test
1 μ L 100mM phenol (being dissolved in DMSO), 1 μ L200mM 4- ammonia are sequentially added into the simulation enzyme solutions of 100 μ L
Interval time is added as 5s in base antipyrine aqueous solution and 1 μ L 200mM aqueous hydrogen peroxide solution, sample, and recording wavelength is
The trap (indoxyl amino-antipyrine dyestuff) of 510nm changes with time, and passes through indoxyl amino-antipyrine dyestuff
Molar extinction coefficient, calculate H2O2Rate of reduction and phenol oxidation rate.
Wherein, phenol is oxidized with the rate of 60nM/s.
Embodiment 3
The present embodiment is prepared by the following method self-assembled nanometer material
1 μ L is sequentially added into the MES/MES sodium salt buffer solution (pH 7.0 contains NaCl 50mM) of 90 μ L 50mM
200 μM of G-DNA, the aqueous streptomycin of 8 μ L 2mM and the manganese protoporphyrin (being dissolved in dimethyl sulfoxide) of 100 μM of 1 μ L, sample
It is 10s that time interval, which is added, in product, and 30min is then mixed at 20 DEG C, and reaction obtains analogue enztme, i.e. self-assembled nanometer material.
Wherein, the sequence of G-DNA is GGTAGGCGGCGGTGGCGGCGGAGG, is purified by HPLC or PAGE.
Catalytic activity test
1 μ L 100mM phenol (being dissolved in DMSO), 1 μ L200mM 4- ammonia are sequentially added into the simulation enzyme solutions of 100 μ L
Interval time is added as 5s in base antipyrine aqueous solution and 1 μ L 200mM aqueous hydrogen peroxide solution, sample, and recording wavelength is
The trap (indoxyl amino-antipyrine dyestuff) of 510nm changes with time, and passes through indoxyl amino-antipyrine dyestuff
Molar extinction coefficient, calculate H2O2Rate of reduction and phenol oxidation rate.
Wherein, phenol is oxidized with the rate of 70nM/s.
Embodiment 4
The present embodiment is prepared by the following method self-assembled nanometer material
1 μ L is sequentially added into the MES/MES sodium salt buffer solution (pH 7.0 contains NaCl 50mM) of 90 μ L 50mM
The chitosan molecule (viscosity 200mPas, the aqueous solution containing 1% acetic acid) of 200 μM of G-DNA, 2 μ L 1mg/mL and 1 μ L
100 μM of hemin (being dissolved in dimethyl sulfoxide), it is 10s that time interval, which is added, in sample, is then mixed at 25 DEG C
30min, reaction obtain analogue enztme, i.e. self-assembled nanometer material.
Wherein, the sequence of G-DNA is GGGTTAGGGTTAGGGTTAGGGTTAGGGTTAGGGTT, by HPLC or PAGE
Purifying.
The self-assembled nanometer material of preparation is subjected to Electronic Speculum observation, it is specific as shown in Figure 1.It can see clearly nanometer material
The particle of material, it is widely distributed.
Catalytic activity test
1 μ L 100mM amino-phenol (being dissolved in DMSO), 1 μ L 200mM are sequentially added into the simulation enzyme solutions of 100 μ L
Interval time is added as 5s in 4-AA aqueous solution and 1 μ L 200mM aqueous hydrogen peroxide solution, sample, and recording wavelength is
The trap (indoxyl amino-antipyrine dyestuff) of 510nm changes with time, and passes through indoxyl amino-antipyrine dyestuff
Molar extinction coefficient, calculate H2O2Rate of reduction and amino-phenol oxidation rate.
Wherein, amino-phenol is oxidized with the rate of 100nM/s.
Embodiment 5
The present embodiment is prepared by the following method self-assembled nanometer material
4 μ L are sequentially added into the MES/MES sodium salt buffer solution (pH 7.0 contains NaCl 50mM) of 90 μ L 50mM
25 μM of G-DNA- hemin conjugate, the paromomycin sulfate aqueous solution of 8 μ L 2mM, sample are added time interval and are
10s then mixes 30min at 25 DEG C.
Wherein, the sequence of G-DNA is GGGTTAGGGTTAGGGTTAGGG, purifies by HPLC or PAGE, has alkynes at the end 3'
Base modification；It reacts coupling by click chemistry with the hemin of nitrine modification.
Catalytic activity test
1 μ L 100mM amino-phenol (being dissolved in DMSO), 1 μ L 200mM are sequentially added into the simulation enzyme solutions of 100 μ L
Interval time is added as 5s in 4-AA aqueous solution and 1 μ L 200mM aqueous hydrogen peroxide solution, sample, and recording wavelength is
The trap (indoxyl amino-antipyrine dyestuff) of 510nm changes with time, and passes through indoxyl amino-antipyrine dyestuff
Molar extinction coefficient, calculate H2O2Rate of reduction and amino-phenol oxidation rate.
Wherein, amino-phenol is oxidized with the rate of 100nM/s.
Comparative example 1
The difference of this comparative example and embodiment 4 is only that, does not include the hemin of 100 μM of 1 μ L, is prepared certainly
Assemble nanometer material, and carry out catalytic activity test.
Material prepared by comparative example 1 does not have any catalytic activity.
Comparative example 2
The difference of this comparative example and embodiment 4 is only that, does not include the G-DNA of 200 μM of 1 μ L, self assembly is prepared and receives
Rice material, and carry out catalytic activity test.
For tetramethyl benzidine, substrate conversion number is 0.129s-1, catalytic efficiency 1.76mM-1·s-1。
Comparative example 3
The difference of this comparative example and embodiment 4 is only that, does not include the chitosan molecule of 2 μ L 1mg/mL, is prepared certainly
Assemble nanometer material, and carry out catalytic activity test.
For tetramethyl benzidine, substrate conversion number is 0.94s-1, catalytic efficiency 58.75mM-1·s-1。
The Catalytic data of nano material prepared by embodiment 1 and comparative example 2, comparative example 3 is made into reaction monitoring curve graph,
As shown in Figure 2.
The Applicant declares that the present invention is explained by the above embodiments self-assembled nanometer material and its preparation side of the invention
Method and application, but the invention is not limited to above-mentioned processing steps, that is, do not mean that the present invention must rely on above-mentioned processing step
It could implement.It should be clear to those skilled in the art, any improvement in the present invention, to raw material selected by the present invention
Equivalence replacement and addition, the selection of concrete mode of auxiliary element etc., all fall within protection scope of the present invention and the open scope
Within.
Claims (10)
1. a kind of self-assembled nanometer material, which is characterized in that the raw material of the nano material includes nucleic acid molecules, aminoglycoside
Molecule and metal protoporphyrin.
2. self-assembled nanometer material according to claim 1, which is characterized in that the nucleic acid molecules are DNA or RNA；
Preferably, in DNA, molar ratio shared by guanine deoxyribonucleotide is 10%-100%；
Preferably, in DNA, guanine deoxyribonucleotide number is 4-59；
Preferably, in RNA, molar ratio shared by guanosine ribonucleoside acid is 10%-100%；
Preferably, in RNA, guanosine ribonucleoside acid number is 4-59；
Preferably, the nucleic acid molecules are DNA, and molar ratio shared by guanine deoxyribonucleotide is 50%-100%,
Guanine deoxyribonucleotide number is 19-24.
3. self-assembled nanometer material according to claim 1 or 2, which is characterized in that the aminoglycoside molecule includes
Any one in chitosan, paromomycin sulfate, streptomysin, neomycin, amikacin or tobramycin；
Preferably, the aminoglycoside molecule is chitosan；
Preferably, the deacetylation of the chitosan is 10%-100%, preferably 95%-100%；
Preferably, the viscosity of the chitosan is 100mPas-800mPas, preferably 100mPas-200mPas.
4. self-assembled nanometer material according to any one of claim 1-3, which is characterized in that the metal protoporphyrin packet
It includes in iron-protoporphyrin, Copp, manganese protoporphyrin, nickel protoporphyrin, copper protoporphyrin, ruthenium protoporphyrin, rhodium protoporphyrin or palladium protoporphyrin
Any one；
Preferably, the metal protoporphyrin is iron-protoporphyrin；
Preferably, the iron-protoporphyrin is hemin or hemn, preferably hemin.
5. the preparation method of self-assembled nanometer material described in any one of -4 according to claim 1, which is characterized in that by nucleic acid
After molecule and aminoglycoside molecular mixing, metal protoporphyrin is dissolved in progress self assembly in buffer solution and obtains the self assembly
Nano material.
6. preparation method according to claim 5, which is characterized in that the buffer solution includes NaH2PO4/Na2HPO4It is slow
Rush solution, KH2PO4/K2HPO4Buffer solution, Tris/Tris hydrochloric acid salt buffer solution, HEPES/HEPES sodium salt buffer solution or
Any one in MES/MES sodium salt buffer solution；
Preferably, the pH value of the buffer solution is 3-10；
Preferably, the buffer solution is MES/MES sodium salt buffer solution；PH value is preferably 6.5-7.5.
7. preparation method according to claim 5 or 6, which is characterized in that the aminoglycoside molecular concentration is
0.001mg/mL-1.5mg/mL, preferably 0.02mg/mL-0.1mg/mL；
Preferably, the nucleic acid molecules concentration is 0.1nM-250 μM；
Preferably, the metal protoporphyrin concentration is 0.1nM-150 μM, preferably 100nM-100 μM.
8. the preparation method according to any one of claim 5-7, which is characterized in that the temperature of the self assembly be 4 DEG C-
45 DEG C, preferably 20 DEG C -30 DEG C；
The time of the self assembly is 10s-60min, preferably 10min-30min.
9. a kind of catalyst, which is characterized in that the catalyst includes as self assembly of any of claims 1-4 is received
Rice material；
Preferably, the catalyst has catalyzing hydrogen peroxide activity, catalysed reductive substrate active, chiral peroxide substrate
Selective oxidation activity and chiral reduction substrate selective oxidation activity；
Preferably, the reproducibility substrate includes tetramethyl benzidine, phenol, pyrogallol, dopamine, cresols, amino phenols, two
Any one in nitro o-cresol, naphthols, pentachlorophenol, luminol or 10- acetyl group -3,7- dihydroxy phenoxazine；
Preferably, the chiral reduction substrate includes DOPA enantiomter, N- hydroxyl-arginine enantiomter, smart ammonia
Any one in sour enantiomter or tyrosine derivative enantiomter；
Preferably, the hydrogen peroxide converts number in catalyst surface as 0.5-50s-1；
Preferably, the hydrogen peroxide is 0.5-10mM by the efficiency of catalysis reduction-1·s-1；
Preferably, the reproducibility substrate converts number in catalyst surface as 0.1-20s-1；
Preferably, the reproducibility substrate is 0.5-50 μM by the rate of catalysis oxidation-1·s-1；
Preferably, the enantiomeric excess value of the chiral reduction substrate is 10%-99.9%.
10. self-assembled nanometer material described in any one of -4 is in chemical sensor or biosensor according to claim 1
Application；
Preferably, the chemical sensor is antibiotic molecule sensor；
Preferably, the biosensor is antibody sensor, antigen sensor, Protein sensor, nucleic acid sensor or core
Acid identifies any one in substrate sensor.
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| CN114477097A (en) * | 2022-01-12 | 2022-05-13 | 江南大学 | Method for producing hydrogen peroxide based on porphyrin-based supramolecules through photo-thermal catalysis without sacrificial agent |
| CN114644639A (en) * | 2022-03-11 | 2022-06-21 | 江南大学 | Multi-component nano assembly with chiral optical activity and preparation method thereof |
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| CN114644639A (en) * | 2022-03-11 | 2022-06-21 | 江南大学 | Multi-component nano assembly with chiral optical activity and preparation method thereof |
| CN116328840A (en) * | 2022-07-20 | 2023-06-27 | 南京大学 | Heme-based chimeric oligopeptide-DNA mimic enzyme and application |
| CN115779893A (en) * | 2022-12-19 | 2023-03-14 | 江南大学 | H 2 TPPS-arginine assembly mediated chiral noble metal nano catalyst and preparation method thereof |
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