29 results about "Acetylhomocholine" patented technology

The invention discloses an enzyme biosensor for rapid detection of organophosphorus pesticide dimethoate as well as preparation and application of the enzyme biosensor, and relates to the field of electrochemical sensors. The method comprises the following steps: preparing GR / GCE, preparing AuNPs / GR / GCE, and preparing AChE / AuNPs / GR / GCE. Graphene and gold nanoparticles are compounded to form AuNPs / GR / GCE, the electrochemical impedance of the glassy carbon electrode is remarkably reduced, the effective specific surface area of the glassy carbon electrode is increased, the electron transfer rate of the surface of the glassy carbon electrode is increased, rich binding sites are provided for immobilization of enzyme molecules, the stability of enzyme is improved, and degradation or inactivation of the enzyme in the using process is reduced; acetylcholine esterase is used as a recognition probe, AChE / AuNPs / GR / GCE is constructed on AuNPs / GR / GCE in a self-assembly mode, dimethoate can prevent acetyl thiocholine iodide from being hydrolyzed to generate thiocholine, electrochemical response signals are reduced, and the dimethoate has good electrochemical response signals, so that the organophosphorus pesticide dimethoate residue is rapidly, accurately and safely screened on site.

The invention provides a detection sensor and application thereof, and belongs to the field of pesticide detection. The detection sensor provided by the invention comprises an acetylcholine solution, an acetylcholin esterase solution, a choline oxidase solution, a Cu-BTC dispersion liquid and a solution containing a color developing agent capable of generating a hydroxyl color developing group, or comprises acetylcholine, a color developing agent capable of generating a hydroxyl color developing group and a Cu-BTC-based composite material; the Cu-BTC based composite material is prepared from Cu-BTC, choline oxidase and acetylcholin esterase, wherein the choline oxidase and the acetylcholin esterase are loaded on the Cu-BTC. Acetylcholine is hydrolyzed by acetylcholin esterase to generate choline, the choline is oxidized by choline oxidase to generate H2O2, the H2O2 is oxidized by Cu-BTC to generate. OH, and the. OH can oxidize a color developing agent to generate a colored substance. The organophosphorus pesticide can inhibit the activity of acetylcholin esterase to change the color of the solution, and then quantitative analysis of the organophosphorus pesticide is realized through the absorbance and the absorbance of a standard solution.

The present invention relates to the technical field of plant extracts, and in particular to the use of a ground nasturtium extract in the preparation of an acetylcholinesterase inhibitor. Studies have shown that the ethanol extract of the ground nasturtium and its flower buds (i.e., the ground nasturtium crude extract), the refined extract (RC1-RC6) and the C1-C6 compounds of the present invention have a strong inhibitory effect on acetylcholinesterase, and have good application prospects in the development of drugs for the prevention and treatment of dementia and Alzheimer's disease.

The invention provides a method for rapidly detecting pesticide residues in fruits and vegetables, and belongs to the field of pesticide analysis and detection. The method comprises the following steps: mixing a copper nanoparticle / reduced graphene oxide / chitosan composite material, acetylcholin esterase, choline oxidase and a cross-linking agent glutaraldehyde solution for reaction to obtain a double-enzyme functionalized nano composite material; dispensing a chitosan solution on the surface of a working electrode of a screen-printed electrode, drying to form a film, and covalently modifying the surface of the electrode with the bienzyme functionalized nano composite material through a cross-linking agent glutaraldehyde to obtain a sensing electrode; according to the method, acetylcholine is taken as a substrate, a sensing electrode is used, a chronoamperometry is adopted, inhibition ratios of pesticide standard substance solutions with different known concentrations to electrode response current are detected, and a standard curve between the pesticide concentration and the current inhibition ratio is established; and calculating the pesticide residue concentration in the fruit and vegetable sample to be detected according to the current inhibition ratio of the liquid to be detected and the standard curve. Therefore, the sensitivity and accuracy of pesticide residue detection in fruits and vegetables are improved.

The application provides a kind of gold nanoclusters with polypeptide as ligand and detection method of methyl parathion, the sequence of polypeptide in the gold nanoclusters with polypeptide as ligand is H (S, D) GGFFYCC;The gold nanoclusters with polypeptide as ligand can be used for double signal detection of methyl parathion and detection of methyl parathion in fruits and vegetables.Fluorescence signal detection: gold nanoclusters are added to the test solution, 37 DEG C reaction 30-60 min, detect the fluorescence change at 700 nm.Colorimetric signal: in pH=8.0 buffer solution, different concentrations of methyl parathion, acetylcholinesterase, choline oxidase and acetylcholine are added, 37 DEG C preheating 30-60 min.pH=4.0 buffer solution, gold nanoclusters and TMB are added, 37 DEG C reaction 10 min, 652 nm is measured absorbance.The detection method of the application has high detection sensitivity, strong anti-interference ability, low detection limit, high correlation coefficient of detection curve, good reproducibility in fruit and vegetable detection recovery experiment, and has good practical value in the detection of methyl parathion.

The invention provides a detection sensor and application thereof, and belongs to the field of pesticide detection. The detection sensor provided by the invention comprises acetylcholine, acetylcholin esterase, choline oxidase, two-dimensional Cu-TCPP (Fe) NSs and a color developing agent capable of generating a hydroxyl color developing group. The acetylcholin esterase can hydrolyze acetylcholine to generate choline, the choline is oxidized by the choline oxidase to generate H2O2, the H2O2 is oxidized by the two-dimensional Cu-TCPP (Fe) NSs to generate hydroxyl free radicals (. OH), and the. OH can oxidize a color developing agent capable of generating hydroxyl color developing groups to generate colored substances. When the organophosphorus pesticide exists in the to-be-detected liquid, the color of the solution is light blue or colorless; otherwise, the color of the solution is dark blue. The organophosphorus pesticide can inhibit the activity of acetylcholin esterase, so that the color of the solution is changed in the whole cascade catalytic amplification reaction, and then quantitative analysis of the organophosphorus pesticide is realized through a concentration-absorbance curve of the solution absorbance and the standard solution.

The invention includes a chimeric autoantibody receptor (CAAR) specific for anti-acetylcholine receptor (AChR) B cell receptor (BCR), compositions comprising the CAAR, polynucleotides encoding the CAAR, vectors comprising a polynucleotide encoding the CAAR, and recombinant cells, e.g., T cells comprising the CAAR.

The invention discloses a preparation method of isothiocyanate in papaya seeds, which comprises the following steps: weighing choline, glycerol, sugar and water, heating and stirring uniformly, and taking out when the solid disappears and the solution is colorless or yellowish viscous liquid, so as to obtain a deep eutectic solvent DES; the choline is choline chloride or acetylcholine; weighing pawpaw seeds, crushing the pawpaw seeds, screening out powder through an 80-mesh standard sieve, collecting the powder, weighing the pawpaw seed powder, putting the powder into a reactor, adding the synthesized deep eutectic solvent DES, putting the reactor into a constant-temperature mixing instrument, and carrying out enzymatic reaction, so as to obtain a pawpaw seed enzymolysis product isothiocyanate mixed solution after the reaction is finished. The method is simple in reaction steps, suitable for industrial production and high in reaction yield.

The present application provides a biological product and a method and an application thereof. The biological product is used for the preparation of a therapeutic agent for cancer treatment, and it includes at least one of the recombinant butyrylcholinesterase-albumin (rBChE-albumin) fusion protein and the recombinant butyrylcholinesterase-Fc (rBChE-Fc) fusion protein. The biological product adjusts the balance of acetylcholine and other related metabolic pathways by supplementing at least one of the exogenous rBChE-albumin fusion protein and rBChE-Fc fusion protein to treat cancer. The present invention solves the problem that the extraction and purification of BChE from human serum may not meet the huge demand for cancer treatment. The present application further provides a biological product for neutralizing and degrading nerve agents and organophosphate compounds, hydrolyzing ghrelin, and anti-wrinkle.

The present application relates to a controllable multi-enzyme cascade AChE-HRP-CHO@HNF composite material preparation and plant wearable sensor application, belong to biosensing, enzyme immobilization material and plant in-situ rapid detection field. The composite material is prepared by biomimetic mineralization and layer-by-layer protein crown assembly, takes enzyme-inorganic hybrid nanoflower as carrier, realizes spatial ordered fixation of choline oxidase (CHO), horseradish peroxidase (HRP) and acetylcholinesterase (AChE) in nanostructure, and forms hierarchical distribution multi-enzyme cascade system. The structure can shorten the transmission distance of intermediate product in cascade reaction, improve the cascade catalytic efficiency and pesticide inhibition response sensitivity. Further embedding it in double network hydrogel, a hydrogel sensing disc suitable for plant surface attachment can be prepared, the hydrogel sensing disc has good flexibility, surface adaptability and storage stability, can be used for on-site visual detection of organophosphorus pesticides such as chlorpyrifos, has the characteristics of simple operation and non-invasive detection.

The application discloses a FRET fluorescent probe with dual response to acetylcholinesterase and viscosity, a preparation method and application thereof, wherein the structure of the FRET fluorescent probe is shown in the following formula: The fluorescent probe is based on a fluorescence resonance energy transfer (FRET) mechanism, responds to acetylcholinesterase (AChE) at a short wave of 520 nm, and the detection limit is as low as 0.104 U / L, and responds to viscosity at a long wave of 680 nm, and the wavelength difference is as high as 160 nm, and the dual-channel response has no crosstalk. Cell toxicity test shows that the probe has good biocompatibility, and confocal fluorescence microscopic imaging experiment shows that the probe has good light stability for cells, has the ability of simultaneously monitoring the concentration change of AChE in cells and the viscosity fluctuation of cells, and is suitable for confocal fluorescence imaging for tracing the concentration change of AChE in cells and the viscosity fluctuation.

The invention discloses a multi-target anti-senile dementia activity holothuria scabra-derived lipid compound as well as a separation component and application thereof, and belongs to the field of medicines and functional foods. The preparation method of the holothuria scabra lipid compound comprises the following steps: extracting holothuria scabra total lipid by using a mixed solvent of chloroform and methanol, deoiling by using cold acetone, and then separating and collecting precipitate to obtain the holothuria scabra lipid compound. The holothuria scabra separation component is obtained by carrying out silica gel column chromatography on the holothuria scabra lipid compound, and eluting by taking a mixed solution of dichloromethane, methanol and / or water as an eluent, so as to obtain a primary component with activity after elution; respectively carrying out normal-phase column chromatography on the two components with the best activity, and eluting by using a mixed solvent of ethyl acetate and normal hexane and / or a mixed solvent of dichloromethane and methanol to obtain a secondary component; the holothuria scabra lipid complex and the separated component thereof have remarkable Abeta neurotoxicity resistance, neuroinflammation resistance, intracellular oxidative stress resistance, free radical scavenging and acetylcholin esterase inhibition activity.

The application is suitable for the technical field of pesticide detection, and provides a method for double-signal detection of trace residues of organophosphorus pesticides based on gold nanoscale enzyme, polypeptide is used as a protective agent to synthesize gold nanoscale enzyme, the double properties of fluorescence and peroxidase-like activity of the gold nanoscale enzyme are used, and the enzyme inhibitor method is combined to establish a detection method, and the specific steps are as follows: step one, synthesizing gold nanoscale enzyme and storing for standby; step two, preparing sodium alginate hydrogel blocks, and physically adsorbing gold nanoscale enzyme into the sodium alginate hydrogel through soaking and storing for standby; step three, establishing an acetylcholinesterase / choline oxidase cascade reaction to generate H2O2; step four, based on the H2O2 generated in step three, and combining the prepared sodium alginate hydrogel in step two, realizing rapid visual quantitative detection of OPs, the double-signal output detection result of the application has self-correction, can resist interference, improves detection accuracy, and expands the practical application range of detection.

The application provides an organic phosphorus poisoning relief composite enzyme and application thereof, and belongs to the technical field of biological medicine. The organic phosphorus poisoning relief composite enzyme provided by the application comprises polyethylene glycol modified organic phosphorus hydrolase and butyrylcholine esterase, and the mass ratio of the polyethylene glycol modified organic phosphorus hydrolase to the butyrylcholine esterase is (4-8):(2500-10000). The rat acute organic phosphorus poisoning treatment result shows that the organic phosphorus poisoning relief composite enzyme provided by the application fully plays the efficient catalytic hydrolysis capacity of the enzyme as a catalyst, efficiently removes organic phosphorus and acetylcholine by using the organic phosphorus hydrolase and the butyrylcholine esterase respectively, provides a drug combination for treating the cause for the organic phosphorus poisoning injury first aid, and the treatment effect is significantly better than that of the single drug.

The use of deep eutectic solvents in enzyme-based biosensors improves the robustness of the sensors with regard to storage conditions. For example, phosphotriesterase or acetylcholinesterase enzymes can be used to detect organophosphates using electrochemistry.

The invention discloses an acetylcholin esterase and butyrylcholin esterase double-target inhibitor, which has the following chemical structural formula: wherein R is selected from any one of the following structural formulae;

The application provides a preparation method of metal-doped carbon dots, comprising the following steps: drying an aqueous solution of reactants containing ethylenediaminetetraacetate, Cu 2+ and Fe 3+ , high-temperature pyrolysis, acid washing, water washing, methanol purification and filtering, and drying at 60 DEG C to obtain the metal-doped carbon dots. The phenol measurement steps provided by the application are as follows: mixing a sample solution with a buffer solution, a 4-aminoantipyrine solution and a carbon dot solution, incubating to obtain a solution A, and measuring the absorbance of the solution A at 509 nm for quantitative analysis. The measurement steps of dimethoate provided by the application are as follows: mixing a sample solution with a acetylcholinesterase solution, incubating for half an hour, adding a acetylthiocholine chloride solution and incubating for another half an hour, adding a solution A, and incubating for half an hour before fluorescence measurement. The synthesis method provided by the application is simple, the detection method has high sensitivity and small interference, and is suitable for simultaneous detection of phenol and dimethoate in actual samples.

The invention provides alkaloid 1-(4-hydroxyphenyl)-beta-carboline (PFT217) in Gastrodia elata Blume, a preparation method of the alkaloid 1-(4-hydroxyphenyl)-beta-carboline (PFT217) and application of the alkaloid 1-(4-hydroxyphenyl)-beta-carboline (PFT217) in the field of medicine. Belongs to the technical field of medicine. The PFT217 has remarkable inhibitory activity on acetylcholin esterase and butyrylcholin esterase, and the IC50 (half maximal inhibitory concentration) values of the PFT217 are 24.70 mu M and 0.84 mu M respectively. The alkaloid can be used for preparing medicines for resisting Alzheimer's disease.

The invention provides an HSA fluorescent probe as well as synthesis and application thereof. The fluorescent probe is an enzymatic HSA fluorescent probe and has a structure as shown in a formula (I). The fluorescent probe has high selectivity and high sensitivity, can distinguish HSA from other enzymes (such as chymotrypsin, lipase, protease, beta-galactosidase, pepsin, acetylcholin esterase, butyrylcholine esterase, carboxylesterase, trypsin, DNA enzyme, alkaline phosphatase, lysozyme and the like), and has the detection limit as low as 0.27 mu g / mL, so that the fluorescent probe has the advantages of high selectivity, high sensitivity, high sensitivity and the like. The fluorescent probe provided by the invention can realize qualitative and quantitative detection of HSA in serum, urine or cells.

The invention relates to a large Stokes shift cholinesterase near-infrared fluorescent probe and a construction method thereof.Quinoline is used as a core skeleton, and a conjugated system is constructed through a D-pi-A electronic regulation strategy: aryl, alkenyl or amido is introduced to the 4th site / 6th site to regulate the emission wavelength; the construction method comprises the following steps: synthesizing a fluorophore main skeleton through a Wittig reaction / Heck reaction, connecting a recognition unit through a substitution reaction, and introducing the electron-withdrawing group through a Najing condensation reaction. Stokes shift breaks through 100 nm, and excitation light interference can be inhibited. According to the probe, a recognition unit simulating acetylcholine / butyrylcholine'dumbbell-shaped 'conformation is covalently connected to a quinoline nitrogen atom through a benzyl bromide group, the recognition unit comprises an aromatic cavity adaptive to AChE / BChE substrate pocket hydrophobicity and an active site for catalyzing ester bond hydrolysis, an enzymatic reaction triggers intramolecular electron rearrangement and C-N bond breakage, a fluorescence quenching state is relieved, and the quinoline nitrogen atom can be detected. And "off-on" type signal conversion is realized.

The invention relates to a paper-based sensor, a preparation method of the paper-based sensor and application of the paper-based sensor in detection of alternariol, and belongs to the technical field of food detection. The paper-based sensor comprises a first protective layer, an immobilization layer, a substrate layer, a color development layer and a second protective layer, the surfaces of the first protective layer and the second protective layer are coated with hydrophobic materials; central areas of the immobilization layer, the substrate layer and the color development layer are coated with acetylcholin esterase, acetylthiocholine and nano-enzyme respectively to form hydrophilic reaction areas, and non-hydrophilic reaction areas are coated with hydrophobic materials to form hydrophobic isolation areas; the first protection layer, the immobilization layer, the substrate layer, the color development layer and the second protection layer are folded according to a preset sequence, and the hydrophilic reaction areas of the immobilization layer, the substrate layer and the color development layer are mutually overlapped after folding. The paper-based sensor is remarkably improved in the aspects of detection sensitivity, stability and convenience.

The use of deep eutectic solvents in enzyme-based biosensors improves the robustness of the sensors with regard to storage conditions. For example, phosphotriesterase or acetylcholinesterase enzymes can be used to detect organophosphates using electrochemistry.

The invention provides a ternary complex dual-mode probe and an acetylcholin esterase activity and enzyme inhibitor detection method thereof. The ternary complex dual-mode probe is composed of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system, a manganese dioxide nanoflower-CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) activated ssDNA (ssDNA) (MnO2 NFs-ssDNA) and internal reference fluorescence. When a target to be detected is acetylcholin esterase (AChE), hydrolysis of an acetylthiocholine substrate can be catalyzed, a thiocholine product can be generated, MnO2 NFs can be decomposed to release ssDNA, the trans-cleavage activity of Cas12a can be activated, fluorescence report nucleic acid can be cleaved, a fluorescence signal can be generated, and a more accurate ratio fluorescence signal can be output by taking internal reference fluorescence added into a system as an internal standard; the decomposition of the MnO2 NFs reduces the oxidase-like activity of the MnO2 NFs, and the colorimetric signal for catalyzing the color development of 3, 3 ', 5, 5'-tetramethyl benzidine is reduced; therefore, the higher the AChE activity is, the larger the yield of the thiocholine is, the more thorough the decomposition of the MnO2 NFs is, the stronger the fluorescence signal is, and the weaker the colorimetric signal is. The enzyme inhibitor organophosphorus pesticide can inhibit generation of thiocholine, and then an opposite experiment result appears.

The present invention provides a class of HSA fluorescent probes and their synthesis and applications. The fluorescent probe is an enzymatic HSA fluorescent probe having a structure represented by formula (I). The fluorescent probe of the present invention has high selectivity and high sensitivity, capable of distinguishing HSA from other enzymes (such as chymotrypsin, lipase, protease, β-galactosidase, pepsin, acetylcholinesterase, butyrylcholinesterase, carboxylesterase, trypsin, DNA enzyme, alkaline phosphatase, lysozyme, etc.), with a detection limit as low as 0.27 μg / mL. Therefore, the fluorescent probe of the present invention can achieve qualitative and quantitative detection of HSA in serum, urine, or cells.

The invention belongs to the field of analytical chemistry, and particularly relates to molybdenum disulfide modulated iron monatomic bionic nano-enzyme as well as a preparation method and application thereof. The bionic nano-enzyme is in a hollow nano-rod shape, a carbon base anchored with iron monatomic atoms is uniformly doped with molybdenum disulfide lamellae, the iron monatomic atoms are anchored through nitrogen uniformly doped on the carbon base, the molybdenum disulfide lamellae are composed of 1T-phase molybdenum disulfide, a FeN5-MoS2 active center is formed, catalytic activity is achieved, and the carbon base and the molybdenum disulfide lamellae can be used for preparing the nano-enzyme. The heme b5-molybdenum cofactor is similar to the heme b5-molybdenum cofactor of natural sulfite oxidase, and has efficient peroxidase-like activity and good stability. Colorimetric signals are generated on the basis that the activity of iron atoms is affected by acetylcholine hydrolysate, a cascade detection platform with acetylcholin esterase as a marker is constructed, and sensitive detection of insecticide acetamiprid and distinguishing of different natural alkaloids are achieved. In order to adapt to convenient real-time detection in a complex environment, a paper-based colorimetric sensing platform is combined with a paper base, the residual content of acetamiprid is read through a mobile phone colorimetric program, and the method has the advantages of low cost, simplicity in operation, good specificity, high sensitivity and the like.

The application discloses a double-enzyme cascade amplification fluorescence sensor based on electrostatic driving and a preparation method and application thereof, and the nano-enzyme fluorescence sensor is composed of a probe Azo-Bodipy 685, choline oxidase (CHO) and acetylcholine chloride (ACh). Based on the electrostatic adsorption principle, the application proposes an electrostatic complex co-immobilization-enzyme cascade reaction synergistic strategy, constructs a new electrostatic driving fluorescence sensor, utilizes the electrostatic effect of the positive charge of the quaternary ammonium salt of ACh and the negative charge of the surface of CHO to construct a nano-scale space limited enzyme reaction microenvironment on the surface of a specific fluorescent probe Azo-Bodipy 685. When an organophosphorus pesticide inhibits acetylcholine esterase (AChE), the decrease of the generation of choline chloride leads to the decrease of the amount of hydrogen peroxide generated by CHO catalysis, and the fluorescence signal intensity is weakened, and a series of fluorescence intensity changes can realize the detection of AChE activity and the determination of the content of organophosphorus pesticide residues.

This invention discloses a method for detecting acetylcholinesterase based on a cobalt-doped graphitic carbon nitride photoresponsive nanozyme, comprising the following steps: first, preparing a Co / g-C3N4 nanozyme; then, mixing acetylcholinesterase with buffer and acetylthiocholine, incubating, and subsequently adding buffer, Co / g-C3N4, and TMB; irradiating with a xenon lamp, centrifuging, and recording the ultraviolet-visible absorption spectrum of the supernatant. This method uses a photoresponsive nanozyme, eliminating the need for unstable hydrogen peroxide, and is extremely simple to operate. The catalytic colorimetric reaction of the photoresponsive nanozyme can be precisely controlled by switching the light source on and off, resulting in more accurate detection results. It also provides guidance for the preparation of highly catalytically active g-C3N4-based photoresponsive nanozymes.

The invention discloses a method for directionally screening probiotics capable of producing acetylcholine, and belongs to the technical field of microorganisms. On the basis of bioinformatics analysis of a choline acetyltransferase related gene sequence and a conserved protein structural domain in an acetylcholine synthesis route, a protein structural domain Acetyltransf17 (IPR041380) which is not previously represented and possibly participates in synthesis of acetylcholine is excavated; the function of the gene is further verified through gene knockout and fermentation tests of the strain. The protein structural domain gene is used as a screening target, whether the strain has the capability of producing acetylcholine or not can be confirmed by analyzing the genome of a to-be-detected strain, and compared with a traditional screening method based on a fermentation test, the efficiency can be remarkably improved when the marker is applied to specific screening and identification of the strain.

The invention discloses a method for detecting acetylcholin esterase based on cobalt-doped graphite-phase carbon nitride photoresponse nano-enzyme, which comprises the following steps: firstly preparing Co / g-C3N4 nano-enzyme, then mixing acetylcholin esterase with a buffer solution and acetylthiocholine, incubating, sequentially adding the buffer solution, Co / g-C3N4 and TMB, irradiating with a xenon lamp, centrifuging, and detecting the acetylcholin esterase based on the cobalt-doped graphite-phase carbon nitride photoresponse nano-enzyme. Taking supernate and recording an ultraviolet-visible absorption spectrum. The method adopts the photoresponse nano-enzyme, does not need to use unstable hydrogen peroxide, and is extremely simple and convenient to operate. The catalytic chromogenic reaction of the photo-responsive nano-enzyme can be accurately regulated and controlled by switching on and off the light source, so that the detection result is more accurate. And meanwhile, guidance is provided for preparation of the g-C3N4-based photoresponse nano enzyme with high catalytic activity.