CN114441458B - Application of ZIF material in inhibition of mimic enzyme - Google Patents
Application of ZIF material in inhibition of mimic enzyme Download PDFInfo
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- 108090000790 Enzymes Proteins 0.000 title claims abstract description 21
- 230000003278 mimic effect Effects 0.000 title claims abstract description 17
- 239000000463 material Substances 0.000 title claims abstract description 8
- 230000005764 inhibitory process Effects 0.000 title description 15
- YAGCJGCCZIARMJ-UHFFFAOYSA-N N1C(=NC=C1)C=O.[Zn] Chemical compound N1C(=NC=C1)C=O.[Zn] YAGCJGCCZIARMJ-UHFFFAOYSA-N 0.000 claims abstract description 46
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 11
- XYHKNCXZYYTLRG-UHFFFAOYSA-N 1h-imidazole-2-carbaldehyde Chemical compound O=CC1=NC=CN1 XYHKNCXZYYTLRG-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000003112 inhibitor Substances 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 8
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- 108040007629 peroxidase activity proteins Proteins 0.000 claims 1
- 238000006555 catalytic reaction Methods 0.000 abstract description 9
- 230000003197 catalytic effect Effects 0.000 abstract description 7
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 abstract description 3
- 239000002086 nanomaterial Substances 0.000 abstract description 3
- 239000004246 zinc acetate Substances 0.000 abstract description 3
- 238000002835 absorbance Methods 0.000 description 11
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- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 8
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- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 2
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- 244000063299 Bacillus subtilis Species 0.000 description 1
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
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- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
Abstract
The invention belongs to the field of nano materials and catalysis, and particularly relates to an application of a ZIF material in inhibiting mimic enzyme. Use of nanoscale ZIF-90 in inhibiting the catalytic activity of a mimetic enzyme. The imidazole-2-formaldehyde and zinc acetate react at normal temperature to generate the ZIF-90, and the prepared nanoscale ZIF-90 is found to have excellent performance of inhibiting the catalysis of mimic enzymes. The ZIF-90 prepared by the invention has good stability and biocompatibility, and has wide application prospect in the aspect of inhibiting the catalysis of mimic enzyme.
Description
Technical Field
The invention belongs to the field of nano materials and catalysis, and particularly relates to an application of a ZIF material in inhibiting mimic enzyme.
Background
The Metal Organic Framework (MOF) is composed of metal ions and organic ligands, so that a two-dimensional or three-dimensional coordination polymer is formed, and the Metal Organic Framework (MOF) has the advantages of large porosity, large surface area, adjustable structure and wide functions. The above advantages make MOFs applicable to gas storage, separation, catalysis and electrodes. In recent years, some biocompatible MOFs have also been used in the biosensing and medical fields. Among them, zeolite Imidazole Frameworks (ZIFs) are a class of MOF materials based on transition metals and imidazole linkers, which have chemical stability under extreme biological conditions in addition to permanent voids, adjustable pore sizes and geometries, which make ZIFs have great application potential in the field of building biosensors. Zeolite imidazole ester-90 (ZIF-90) is composed of Zn 2+ Is self-assembled with imidazole-2-formaldehyde, has good stability and biocompatibility, and can be used for constructing a biosensor, serving as a drug transport carrier and the like.
Mimic enzymes are a class of nanomaterials synthesized by organic chemistry with enzyme-like properties. The mimic enzyme has the advantages of low preparation and storage cost, flexible structural design, adjustable catalytic activity and the like, so that the mimic enzyme is more and more concerned by people. In recent years, how to controllably regulate the activity of a mimetic enzyme has become a new problem, and studies have found that the catalytic activity of a mimetic enzyme can be inhibited by cysteine, o-xylene, polyphenol, metal nanoparticles, etc., but ZIFs have not been mentioned as inhibitors affecting the catalytic activity of enzyme mimetics.
Disclosure of Invention
The invention aims to provide an application of a ZIF material in inhibiting mimic enzyme aiming at the current technical problems.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the application of the ZIF material in inhibiting the mimic enzyme and the application of the nanoscale ZIF-90 in inhibiting the catalytic activity of the mimic enzyme.
The application of the nanoscale ZIF-90 as an inhibitor of catalytic activity of an enzyme mimic.
The inhibitor can inhibit MoS 2 Is used for the application of the peroxidase-like activity.
ZIF-90 and several other inhibitors are added to PBS buffer containing 3, 5-Tetramethylbenzidine (TMB) and H 2 O 2 Adding MoS into the system 2 After the reaction, the supernatant was collected and absorbance was recorded.
Further, 200. Mu.L of ZIF-90 (40 mM), BQ (40 mM), TA (40 mM), cysteine (40 mM), glutathione (40 mM) were added to 1300. Mu.L of PBS buffer (10 mM, pH=7.4) containing 0.8mM TMB and 0.25mM H, respectively 2 O 2 Adding MoS into the system 2 (500μL,2mg mL- 1 ) After 5min of reaction, the supernatant was taken and its UV-visible absorption spectrum in the range of 520nm to 770nm was recorded. Blank contains 0.8mM TMB and 0.25mM H to 1500. Mu.L 2 O 2 To PBS buffer (10 mM, pH=7.4) was added MoS 2 (500μL,2mg mL -1 )。
The inhibition of MoS by inhibitors 2 ATP is added to the peroxidase-like activity system to inactivate the inhibitor inhibition.
The inhibitor is used for qualitatively and/or quantitatively detecting ATP content in biological samples, so that inhibition is inactivated.
Further, ZIF-90 inhibition of MoS 2 Oxidized TMB changes color, and the system is colorless. Adding a biological sample into the system, and firstly cracking ZIF-90 skeleton by extracellular free ATP, wherein part of ZIF-90 loses inhibition and activates MoS 2 Oxidized TMB turns blue. Then adding antibacterial peptide to lyse cells, and intracellular ATP continues to lyse ZIF-90, thereby further accelerating MoS 2 And oxidizing TMB, and utilizing the absorbance response of the system to realize ATP detection.
Further, 1000. Mu.L of E.coli solution (OD 600 Add ZIF-90 (200 μl,1mg mL) to =1) -1 ) And TMB (80. Mu.L, 20 mM) and H were added 2 O 2 (20. Mu.L, 25 mM) and finally adding MoS 2 (500μL,2mg mL -1 ) After 5min of reaction, the supernatant was taken and absorbance at 652nm was measured to determine extracellular ATP content. Subsequently, an antibacterial peptide solution (200. Mu.L, 10mg mL) was added -1 ) Decomposing bacteria. After an additional 5min incubation, absorbance measurements were performed. Intracellular ATP concentrations were analyzed by the difference in absorbance response of the solutions before and after the antimicrobial peptide addition.
The nanoscale ZIF-90 is prepared by passing Zn 2+ Reacting with imidazole-2-formaldehyde in DMF solution at normal temperature, and self-assembling to obtain ZIF-90 of 100-200 nM.
The invention has the beneficial effects that:
the imidazole-2-formaldehyde and zinc acetate react at normal temperature to generate the ZIF-90, and the prepared nanoscale ZIF-90 is found to have excellent performance of inhibiting the catalysis of mimic enzymes. The ZIF-90 prepared by the invention has good stability and biocompatibility, and has wide application prospect in the aspect of inhibiting the catalysis of mimic enzyme.
The prepared nanoscale ZIF-90 is utilized to construct a biosensor, and TMB is used as an active substrate for the biosensor, so that the prepared biosensor signal is researched. The substrate TMB can be modified with MoS having the desired peroxidase-like activity 2 Catalysis produces a blue product. ZIF-90 pair MoS 2 The catalytic oxidation TMB has a strong inhibition effect. ATP targets ZIF-90 to decompose, so that the inhibition effect of ZIF-90 disappears, thereby recovering MoS 2 And (3) oxidizing TMB. The constructed sensor has good detection effect and selectivity, and the ATP detection limit is 0.047 mu M.
Drawings
FIG. 1 is an XRD pattern (A), FT-IR spectrum (B) and TEM pattern (C, D) of ZIF-90 synthesized products provided by examples of the invention;
FIG. 2 is a graph showing inhibition of MoS by ZIF-90 and other radical scavengers provided in an embodiment of the present invention 2 A solution ultraviolet-visible absorption spectrum (a) and an inhibition ratio (B) of the oxidized TMB;
FIG. 3 is a graph of the standard curve of the sensor for detecting ATP at different concentrations according to the embodiment of the present invention;
FIG. 4 is a diagram showing the detection of intracellular and extracellular ATP in different bacteria by a biosensor according to an embodiment of the present invention.
Detailed Description
The following description of the embodiments of the present invention is further provided in connection with the accompanying examples, and it should be noted that the embodiments described herein are for the purpose of illustration and explanation only, and are not limiting of the invention.
Example 1:
preparation of nanoscale ZIF-90:
to 2mL of imidazole-2-carbaldehyde in DMF (0.05M) was added dropwise 1mL of zinc acetate in DMF (0.05M) with stirring. Stirring for 15min, centrifuging and washing the ZIF-90 nanoparticles obtained by the reaction with DMF, ultrapure water and absolute ethanol 10000r/min for 3 times, and adding 1mg mL -1 Is dispersed in ultrapure water and stored for standby.
Successful synthesis of ZIF-90 crystals was confirmed by XRD, FT-IR and TEM analyses, as shown in fig. 1A, the prominent XRD peaks of ZIF-90 structure were completely matched with the standard pattern obtained by simulation, indicating successful synthesis. In addition, the bonding properties and purity of the synthesized ZIF-90 structure were studied by FTI-R, as shown in FIG. 1B. The result shows that ZIF-90 is 2925cm -1 And 2854cm -1 Aliphatic C-H vibration with imidazole ring at 1678cm -1 The energy band at this point provides evidence for the presence of aldehyde groups in the ZIF-90 structure. 600-1500cm -1 The peak at which corresponds to stretching or bending of the entire ring, 421cm -1 The peak at which corresponds to Zn-N stretching. TEM was also used to characterize the structural morphology of ZIF-90, as shown in fig. 1, C, D, ZIF-90 has uniform nano-size, regular shape,the particle size is between 150nm and 200 nm. The above analysis confirmed the successful synthesis of the ZIF-90 structure.
Example 2:
nanoscale ZIF-90 inhibits the mimic enzyme MoS 2 Catalysis:
200. Mu.L ZIF-90 (40 mM), BQ (40 mM), TA (40 mM), cysteine (40 mM), glutathione (40 mM) were added to 1300. Mu.L PBS buffer (10 mM, pH=7.4) containing 0.8mM TMB and 0.25mM H, respectively 2 O 2 Adding MoS into the system 2 (500μL,2mg mL -1 ) After 5min of reaction, the supernatant was taken and its UV-visible absorption spectrum in the range of 520nm to 770nm was recorded. Blank contains 0.8mM TMB and 0.25mM H to 1500. Mu.L 2 O 2 To PBS buffer (10 mM, pH=7.4) was added MoS 2 (500μL,2mg mL -1 )。
The ultraviolet-visible absorption spectrum of the solution after the reaction is shown in fig. 2A, and compared with a blank group without the antioxidant, the absorbance of each group of samples is reduced to a certain extent, but the absorbance of the samples added with ZIF-90 is obviously lower than that of other inhibitors. The inhibition of the samples was calculated using the following formula using the absorbance of the solution at 652nm, and as shown in FIG. 2B, the inhibition of ZIF-90 was 74.47%.
Example 3:
the biosensor was constructed based on ZIF-90 inhibition performance for use in a range of ATP concentrations.
ZIF-90 (200. Mu.L, 1mg mL) was used for a range of ATP concentrations -1 ) 1300 μl of PBS buffer (10 mM, pH=7.4) containing 0.8mM TMB and 0.25mM H was added 2 O 2 Then add MoS 2 (500μL,2mg mL -1 ) Finally, ATP standard solution (10 mM) is added, the ATP concentration of the system is respectively regulated to be 0, 0.1, 0.5, 1, 5, 10, 30, 50, 80 and 100 mu M, the shaking reaction is carried out for 5min, and the absorbance of the supernatant at 652nm is measured.
As shown in FIG. 3, the sensor has a good linear relationship in the range of 0.1 to 100. Mu.M by analyzing the relationship between the ATP concentration and the absorbance at 652 nm. The ATP detection limit was 0.047. Mu.M (S/N=3), and the sensitivity was very high.
Example 4:
the biosensor is constructed based on ZIF-90 inhibition performance and applied to ATP concentration detection in different bacterial samples.
Gram-negative E.coli and P.aeruginosa were selected and the performance of the gram-positive Staphylococcus aureus, bacillus subtilis and P.mononucleosis verification sensors to detect ATP in biological samples. To 1000. Mu.L of bacterial solution (OD 600 Add ZIF-90 (200 μl,1mg mL) to =1) -1 ) And TMB (80. Mu.L, 20 mM) and H were added 2 O 2 (20. Mu.L, 25 mM) and finally adding MoS 2 (500μL,2mg mL -1 ) After 5min of reaction, the supernatant was taken and absorbance at 652nm was measured to determine extracellular ATP content. Subsequently, an antibacterial peptide solution (200. Mu.L, 10mg mL) was added to the solution -1 ) Decomposing bacteria. After an additional 5min incubation, absorbance measurements were performed. Intracellular ATP concentrations were analyzed by comparing the difference in absorbance responses of the solutions before and after M-AMP release.
As shown in FIG. 4, the detection results are shown, and extracellular ATP and intracellular ATP of all samples can be effectively detected by the sensor, so that the sensor can be constructed to optically detect the intracellular ATP content and intracellular ATP content in biological samples.
Claims (1)
1. An application of a ZIF material in inhibiting mimic enzyme, which is characterized in that: nanoscale ZIF-90 inhibits MoS 2 Is used for the activity of the peroxidase;
the nanoscale ZIF-90 is prepared by passing Zn 2+ Reacting with imidazole-2-formaldehyde in DMF solution at normal temperature, and self-assembling to obtain ZIF-90 of 100-200 nM;
the inhibitor is used for qualitatively and/or quantitatively detecting ATP in biological samples.
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