35 results about "Cholesterol oxidase" patented technology

The invention discloses a tumor cholesterol metabolism regulation microneedle patch and a preparation method, the microneedle patch comprises a needle tip and a backing which are connected, the needle tip comprises a needle tip body and nanoparticles loaded on the needle tip body, the nanoparticle is a manganese ion-doped organic metal framework, the outer layer of the manganese ion-doped organic metal framework is modified with a targeting agent, cholesterol oxidase and superoxide dismutase are carried on the manganese ion-doped organic metal framework, the targeting agent can target a CD44 receptor, and the organic metal framework can carry drugs. The targeted drug can enter tumor cells in a targeted mode, superoxide dismutase catalyzes superoxide anions overexpressed in the tumor cells to generate H2O2 and O2, O2 needed by catalysis is provided for cholesterol oxidase, cholesterol at the tumor site is consumed by the cholesterol oxidase, accumulation of 7-DHC is reduced, meanwhile H2O2 can be generated, and the cholesterol oxidase can be used for catalyzing the tumor site. H2O2 is catalyzed by manganese ions through a Fenton-like reaction to generate OH to induce tumor cells to generate ferroptosis, so that ferroptosis-immune synergistic treatment is realized.

A cholesterol oxidase-loaded biological material is a nanometer material and is characterized by comprising polyacrylic acid, cobalt oxyhydroxide and cholesterol oxidase, cobalt oxyhydroxide is adsorbed on polyacrylic acid to form a compound, and cholesterol oxidase is loaded on the compound. The size of the nano-biological material is about 100-150 nm, the nano-biological material is combined with ultrasound to obtain multiple nano-enzyme activities with enhanced effects and more excellent capabilities of generating active oxygen and relieving hypoxia, and a lipid raft structure on a cell membrane is damaged by the loaded cholesterol oxidase, so that cell migration is inhibited, and the effect of resisting tumor metastasis is achieved.

The invention discloses a graphene-based microneedle sensing electrode preparation method and a blood fat detection system. The method comprises the following steps: preparing a microneedle array, preparing a graphene structure on the surface of the microneedle array by utilizing a chemical vapor deposition technology, and modifying cholesterol oxidase on the surface of the microneedle array; and coating the microneedle array with a water-soluble polymer to obtain the microneedle sensing electrode. According to the application, a graphene nanostructure is constructed on the surface of the microneedle, and cholesterol oxidase is fixed on the surface of the electrode, so that a great specific surface area and high electron mobility of the sensor are realized, the biocompatibility and detection precision of the sensor are improved, and accurate and convenient subcutaneous in-situ detection of cholesterol is realized. The method can be widely applied to the technical field of biomedical engineering micro devices.

The application discloses a marine-derived cholesterol oxidase and a preparation method and application thereof, and relates to the technical field of biology. The amino acid sequence of the marine-derived cholesterol oxidase (SkChOx) is shown in SEQ ID NO. 1 or 3, the specific enzyme activity of the marine-derived cholesterol oxidase can reach 31.9 U / mg, the marine-derived cholesterol oxidase has excellent anti-reverse performance, maintains high stability in the pH range of 5.0-10.0, has good temperature tolerance, organic solvent tolerance and metal ion tolerance, and exhibits unique salt activation effect and salt tolerance. The enzyme is compounded with dithiothreitol (DTT) and nonylphenol polyoxyethylene ether (NP-40) to form a liquid enzyme preparation, and after incubation at 37 DEG C for 7 days under accelerated conditions, the catalytic activity of the liquid enzyme preparation still remains about 87%, the liquid enzyme preparation has excellent long-acting storage stability, and has wide application prospects in the fields of clinical diagnosis, food physical and chemical inspection, biosensor development and industrial preparation of steroid drugs.

The invention provides a rapid detection kit and a rapid detection method for phytosterol, and belongs to the field of food detection.The rapid detection kit for phytosterol comprises a phytosterol standard substance, a standard substance diluent, a substrate solution, an enzyme mixed solution, a stop solution and an elisa plate; the standard substance diluent comprises isopropanol or dimethyl sulfoxide; the substrate solution is prepared from TMB (Tetramethylbenzidine) and a substrate buffer solution; the enzyme mixed solution is a mixed solution of a cholesterol oxidase solution and a horse radish peroxidase solution. By optimizing the composition of each working reagent in the kit, the sensitivity and accuracy of phytosterol detection are remarkably improved. Meanwhile, the rapid detection method provided by the invention is simple and convenient to operate, short in detection time and suitable for on-site rapid detection.

The invention discloses a compound enzyme solution for improving total cholesterol test current, which is prepared from the following components in percentage by weight: 20 to 200mM of buffer solution, 0.5 to 200mg / mL of cholesterol oxidase, 0.1 to 100mg / mL of cholesterol esterase, 0.1 to 25mg / mL of peroxidase, 1 to 6 percent of potassium ferricyanide, 0.1 to 8 percent of glucoside and the balance of water. And the cellulose accounts for 0.01%-2%. When a whole blood sample is tested, the compound enzyme liquid of lipoprotein can be quickly decomposed, so that the test current is improved, and the test time is shortened.

The present invention relates to the technical field of biochemistry, in particular to a composite enzyme solution for total cholesterol detection, a test paper and a detection system. The composite enzyme solution for total cholesterol detection comprises cholesterol oxidase, cholesterol esterase, peroxidase, an electron mediator, a surfactant composition, a thickening agent, a protective agent, a metal inorganic salt and a buffer solution, wherein the surfactant composition comprises one of or a combination of at least two of surfactants with an HLB>23 and one of or a combination of at least two of surfactants with an HLB of 12-14. The composite enzyme solution can be used for detecting total cholesterol by means of an electrochemical method, exhibiting fast detection speed, high sensitivity, and high accuracy.

A method of treating a cancer (e.g., a colorectal cancer, a liver cancer, a breast cancer, or a lung cancer) in a subject in need thereof by administering to the subject a therapeutically effective amount of (i) a bacterium that converts intestinal cholesterol to 4-cholesten-3-one (4-C-3) or (ii) 4-C-3. Also disclosed is a recombinant Escherichia coli TOP10 bacterium comprising a nucleic acid encoding cholesterol oxidase (CHOX), which is capable of converting cholesterol to 4-C-3 and can be used to treat gastrointestinal tract cancers including a colorectal cancer or lung cancer.

The invention relates to the technical field of nano-drugs, in particular to a nano-drug for treating triple negative breast cancer as well as a preparation method and application of the nano-drug. The nano-drug takes hyaluronic acid modified hollow mesoporous copper zinc sulfide nanoparticles as a carrier, and cholesterol oxidase is loaded in the nano-drug. The preparation method adopts a one-pot wet chemical method, and a carrier with a regular structure is formed based on a Kirkendall effect. The nano-drug disclosed by the invention can be delivered to a tumor site in a targeted manner, is rapidly degraded in a weak acid microenvironment, synchronously releases cholesterol oxidase, metal ions with Fenton-like activity and H2S gas, and synergistically sensibilizes ferroptosis and efficiently eradicates tumor stem cells by triggering a cascade reaction of cholesterol depletion-lipid raft destruction-lipid peroxidation storm; a brand new treatment scheme is provided for solving the problems of drug resistance and relapse of the triple negative breast cancer.

The present application constructs a fusion protein by utilizing the specific affinity adsorption property of a carbohydrate binding module (CBM) to cellulose, the fusion protein comprising a cholesteric oxidase and a carbohydrate binding module (CBM), and then uses nanocellulose membranes as the immobilization carrier of the fusion protein for preparing a biosensor element, and the obtained biosensor element can exhibit good catalytic properties and detection stability.

The present invention relates to an electrochemical sensor for multiple detection, a preparation method thereof, and an application thereof, and belongs to the field of biological detection technology. The present invention provides an electrochemical sensor for multiple detection, wherein the reference electrode, the working electrode, and the counter electrode of the sensor are sequentially modified with a GOD-HRP-CS-MB layer, a LOD-HRP-FMN layer, and a COD-CEH-HRP-β-CD layer from bottom to top, wherein the components of the GOD-HRP-CS-MB layer include horseradish peroxidase, methylene blue, glucose oxidase, and chitosan, the components of the LOD-HRP-FMN layer include lactate oxidase and flavin mononucleotide, and the components of the COD-CEH-HRP-β-CD layer include β-cyclodextrin, ferrocene, cholesterol oxidase, horseradish peroxidase, and cholesterol esterase. The sensor can simultaneously measure the concentrations of glucose, lactate, and cholesterol in a sample to be tested.

The invention discloses a medical composite material for synergistically regulating cholesterol and oxidative stress. The material comprises a Fe2-N6 / NC carrier, cholesterol oxidase loaded on the Fe2-N6 / NC carrier and a hyaluronic acid targeting layer modified on the surface, the carrier is a nitrogen-doped carbon material loaded with iron double-atom active sites, the distance between adjacent iron atoms is 0.311 + / -0.009 nm, and the iron content is 0.393 wt%; the average particle size of the composite material is 242.5 nm, and the Zeta potential of the composite material is-23.71 mV; the preparation method comprises the following steps: synthesizing Fe2-N6 / NC through mechanochemical-assisted in-situ MOF packaging, ultrasonically loading cholesterol oxidase and modifying with hyaluronic acid. The material has cholesterol degradation activity (the degradation rate is 33.94%) and active oxygen scavenging activity (the scavenging rate is 78.8%), and can be used for preparing an implant material for joint repair or a medical material for articular cavity injection so as to realize coordinated regulation of osteoarthritis cholesterol metabolism disorder and oxidative stress.

A dry analytical reagent for total cholesterol detection comprises, from top to bottom, an upper supporting layer, a diffusion layer, a reagent layer, a support, and a lower supporting layer, wherein the diffusion layer, the reagent layer, and the support are sequentially arranged in a middle position between the upper supporting layer and the lower supporting layer, the pores at both ends of the upper supporting layer and the lower supporting layer are bonded by the middle supporting layer, and a sample addition hole and a test hole are respectively arranged in the middle position of the upper supporting layer and the lower supporting layer, the diffusion layer is an isotropic porous diffusion membrane with a porosity of 50-85%; the diffusion layer is composed of polystyrene microspheres, an adhesive, and a surfactant, and the particle size of the polystyrene microspheres is 0.1-1 μm; the reagent layer includes a surfactant, cholesterol esterase, cholesterol oxidase, horseradish peroxidase, a colorless dye, a buffer, a stabilizer, and a water-soluble high molecular polymer. The diffusion layer prepared by the present invention has a diffusion path of tens to hundreds of nanometers, which can quickly absorb, evenly distribute, measure, and quickly transmit multiple analytes.

The invention provides a method for synthesizing 4-cholestene-3-ketone based on microorganisms of a cholesterol oxidase truncation. The invention specifically provides a method for producing 4-cholestene-3-ketone, which comprises the following steps: culturing a host cell to obtain a culture product; wherein the host cell can synthesize cholesterol and heterologously express cholesterol oxidase; the cholesterol oxidase truncation is selected from cholesterol oxidase without a complete signal peptide function; and extracting and separating the target product 4-cholestene-3-ketone from the culture product of the host cell. According to the invention, the conversion from cholesterol to 4-cholestene-3-ketone can be directly completed in a living cell environment.

The invention relates to the technical field of biochemistry, in particular to compound enzyme liquid, detection test paper, a detection system and a detection method for total cholesterol detection. The compound enzyme liquid for total cholesterol detection comprises cholesterol oxidase, cholesterol esterase, peroxidase, an electronic mediator and a surfactant composition, and the detection method for total cholesterol detection comprises the following steps: inserting detection test paper into a detection system, putting a to-be-detected sample close to a test paper sample adding port, and adding the to-be-detected sample into the test paper sample adding port; a to-be-detected sample is incubated with the compound enzyme liquid loaded on the electrode layer of the detection test paper, and the electrochemical response is determined by applying potential through a detection system. By using the detection method, the total cholesterol can be detected through an electrochemical method, the detection speed is high, the sensitivity is high, and the accuracy is high.

The invention relates to the technical field of preparation of antitumor drugs, in particular to a drug delivery system based on mesoporous polydopamine and a preparation method and application thereof. The mesoporous polydopamine and the substrate are used for loading hydrophobic simvastatin, cholesterol oxidase and manganese acetate at the same time, and the nano composite material coated with hyaluronic acid has good drug loading capacity, realizes targeted release of drugs, and kills tumor cells and inhibits proliferation of the tumor cells through consumption of the drugs on tumor cell cholesterol.

The invention discloses a high-density lipoprotein cholesterol determination kit and a detection method thereof. The kit comprises a reagent 1 and a reagent 2, the reagent 1 is prepared from a buffer solution, inorganic salt, dextran sulfate, a macromolecular cationic polymer, TOOS, peroxidase, ascorbic acid oxidase and a preservative; the reagent 2 is prepared from a buffer solution, a surfactant 1, 4-aminoantipyrine, cholesterol oxidase, cholesterol esterase and a preservative. The kit has the advantages of being high in accuracy, good in anti-interference performance and the like, automatic detection can be achieved on a full-automatic biochemical analyzer, and compared with a traditional dextran sulfate precipitation method, the content of high-density lipoprotein cholesterol (HDL-C) in a lipid turbid sample can be more accurately detected.

The present invention provides a dry chemical reagent sheet for quantitatively detecting cholesterol and a preparation method thereof, and relates to the field of dry chemical reagent sheets. The dry chemical reagent sheet includes a light-transmitting layer, a reagent layer and a diffusion layer, wherein the reagent layer is arranged above the light-transmitting layer, the diffusion layer is arranged above the reagent layer, the diffusion layer includes a diffusion layer reagent or is formed by a diffusion layer reagent, and the diffusion layer reagent includes a hydrophilic polymer, a surfactant, a particulate matter, cholesterol oxidase, cholesterol ester hydrolase, a peroxidase and a color developer, wherein the particulate matter includes a first particulate matter and a second particulate matter; the number ratio of the first particulate matter to the second particulate matter is 1:(0.25-4), and the difference in particle size between the first particulate matter and the second particulate matter is 1-4 μm. The dry chemical reagent sheet provided by the present invention can quickly and accurately quantitatively detect the cholesterol content in a liquid sample, has good repeatability, a small sample size, is easy to store and transport, has a low cost, and has a wide range of applications.

The invention relates to the technical field of biochemistry, in particular to compound enzyme liquid, detection test paper and a detection system for total cholesterol detection. The compound enzyme liquid for total cholesterol detection comprises cholesterol oxidase, cholesterol esterase, peroxidase, an electronic mediator, a surfactant composition, a thickening agent, a protective agent, metal inorganic salt and a buffer solution. The surfactant composition comprises one or a combination of at least two of surfactants with the HLB being larger than 23 and one or a combination of at least two of surfactants with the HLB being 12-14. The compound enzyme liquid can be used for detecting total cholesterol through an electrochemical method, and is high in detection speed, high in sensitivity and high in accuracy.

This invention discloses a dry chemical reagent for total cholesterol detection, comprising a diffusion layer, a spacer layer, and a reagent layer sequentially overlapped. The diffusion layer includes reflective particles, a binder, a surfactant, cholesterol esterase, and cholesterol oxidase. The reagent layer includes a carrier, horseradish peroxidase, an indicator composition, and a buffer solution. The dry chemical detection reagent of this invention employs a coating method, layering a premixed homogeneous slurry onto a support layer. A spacer layer blocks interlayer interference, and phenolic substances replace photosensitizers as chromogens, combined with a specific ratio of substrate and buffer system. The multi-layered, spacer-free, low-interference vertical detection system constructed by this invention significantly improves the repeatability, accuracy, and stability of the detection reagent.

The application provides a preparation method of a cholesterol-consuming nano reactor and application thereof, and belongs to the technical field of biological medicines. The preparation method comprises the following steps: adding a zinc acetate solution dropwise into a 2-methylimidazole solution to obtain a mixed solution; adding a cholesterol oxidase solution and a new indocyanine green solution dropwise into the mixed solution to perform a reaction, so as to obtain a reaction solution; centrifuging the reaction solution, washing the precipitate, and obtaining ZIF@COD / IR820 nanoparticles; adopting a membrane protein extraction method to obtain Hepa1-6 liver cancer cell membrane fragments and red blood cell membrane fragments; mixing the Hepa1-6 liver cancer cell membrane fragments, the red blood cell membrane fragments and the ZIF@COD / IR820 nanoparticles, and then placing the mixture in a liposome extruder to perform repeated extrusion for several times, so as to obtain HM / ZIF@COD / IR820. The nano reactor prepared by the method can intensify cholesterol consumption of tumor cells, promote tumor cell apoptosis, has excellent homologous targeting, and is beneficial to improving the treatment effect of liver cancer.

To provide an evaluation device that can quickly evaluate an amount of oxidized LDL.SOLUTION: An evaluation device includes: an acquisition unit that acquires information on an amount of change in absorbance per unit time obtained by a measurement method of solubilizing LDL by use of a surface active agent acting on the LDL in a sample, causing peroxidase to act on hydrogen peroxide generated based on an oxidation reaction by a cholesterol oxidase to thereby guide the hydrogen peroxide to a coloring reaction, and measuring a time change in absorbance; and an evaluation unit that evaluates an amount of oxidized LDL in the sample on the basis of the amount of change.SELECTED DRAWING: Figure 4

The invention discloses dry chemical test paper and application thereof. The dry chemical test paper comprises a supporting layer, a blood filtering layer, a filtering layer and a reaction layer, the filtering layer is treated by an R1 reagent, and the reaction layer is treated by an R2 reagent; the R1 reagent comprises a 3-morpholine propanesulfonate buffer solution, polyanions, an Mg < 2 + > reagent and a thickening agent; the R2 reagent comprises a 3-morpholine propanesulfonate buffer solution, a nonionic surfactant, an auxiliary surfactant, a saccharide stabilizer, a protein stabilizer, an auxiliary stabilizer, a preservative, a color developing agent, a chromogen substrate, peroxidase, cholesterol oxidase and cholesterol esterase. The dry chemical test paper can directly detect the low-density lipoprotein cholesterol on the dry chemical film, is not influenced by the concentration of triglyceride in a sample, and is accurate in detection result.

The invention provides a lipoprotein subfraction detection kit and a lipoprotein subfraction detection system, and belongs to the technical field of medical examination.The lipoprotein subfraction detection kit comprises a reagent R1, a reagent R2 and a reagent R3, the reagent R1 comprises a first buffer solution, a surfactant, magnesium chloride, a first protective agent, a first preservative, 4-aminoantipyrine, lipoprotein esterase, cholesterol esterase and ascorbic acid oxidase; the reagent R2 comprises a second buffer solution, a second protective agent, a second preservative, NAD < + > and glycerol dehydrogenase; the reagent R3 comprises a third buffer solution, a third protective agent, a third preservative, cholesterol oxidase, peroxidase and a chromogen substance. The lipoprotein subfraction detection kit provided by the invention is matched with the lipoprotein subfraction detection system provided by the invention, so that synchronous detection of cholesterol and triglyceride can be realized, and further detection of 20 lipoprotein subfractions is realized.

The invention provides a serum total cholesterol (TCH) determination kit, the kit comprises a reagent 1 and a reagent 2, the reagent 1 comprises the following main components: a 3-morpholine-2-hydroxypropanesulfonic acid (MOPSO) buffer solution (pH 7.20), cholesterol esterase (CHER), sodium cholate, a surfactant, a stabilizer and a preservative; and the reagent 2 is prepared from the following main components: a 3-morpholine-2-hydroxy propanesulfonic acid (MOPSO) buffer solution (pH is 7.0), cholesterol oxidase (CHOD), peroxidase (POD), sodium cholate, 10-(N-methyl carbamoyl)-3, 7-bis (dimethylamino) phenothiazine (MCDP), a surfactant, a stabilizer and a preservative. By adding the enzyme protective agent and the high-sensitivity pigment source MCTP, the prepared kit is better in stability and wider in linear range.

The invention discloses a cholesterol oxidase mutant as well as an expression strain and application thereof. According to the invention, cholesterol oxidase from uncultured microorganisms is used as a starting enzyme, a mutation site is obtained through multiple designs of Evcouplings and FoldX, then a recombinant plasmid is constructed, and a mutant gene is obtained. After induced expression of the engineering bacteria containing the mutant plasmids, cholesterol oxidase with improved stability and specific enzyme activity is obtained. Under the condition of 37 DEG C, the specific enzyme activity of the mutant is improved to 4.26 times, and the stability is improved to 1.15 times. The mutant has application value in an enzyme-method cholesterol in-vitro detection diagnostic kit.

The invention relates to the technical field of biomedical materials, in particular to a self-adaptive drug delivery microneedle patch as well as a preparation method and application thereof. Comprising a sulfhydrylation silk fibroin matrix used for preparing a microneedle matrix, a drug-loading liposome encapsulated in the microneedle matrix and cholesterol oxidase loaded in the microneedle matrix, the interior of the sulfhydrylated silk fibroin matrix comprises a three-dimensional space network structure formed by crosslinking disulfide bonds; a lipid-lowering drug is encapsulated in the drug-loaded liposome. According to the microneedle patch, the problem of excessive treatment or insufficient drug effect caused by individual metabolism difference in a traditional drug delivery mode is solved, and real-time synchronization of the drug release rate and the blood fat level change is achieved.

The invention discloses a nano converter for inducing pyroptosis of cells as well as a preparation method and application of the nano converter. The nano converter is prepared from the following raw materials: an energy donor, a photosensitizer, cholesterol oxidase and maleimide-1, 2-distearoyl-sn-glycerol-3-phosphoethanolamine-poly (ethylene glycol) (Mal-DSPE-PEG). In the nano converter, bisoxalate and chlorin e6 form a nano micelle with Mal-DSPE-PEG through hydrophilic and hydrophobic interaction, and cholesterol oxidase reacts with maleimide to be modified on the surface of the nano micelle. The nano converter can effectively convert cholesterol and hydrogen peroxide in tumor senescence cells into singlet oxygen to finally cause pyroptosis of the tumor senescence cells. The nano converter disclosed by the invention has the advantages of universality and high safety due to targeting of metabolic characteristics generally existing in tumor senescence cells. In addition, the nano converter utilizes chemical energy to drive photodynamic therapy, and possibly existing phototoxicity is reduced due to the fact that the nano converter does not depend on an external light source.

The present invention discloses a cholesterol biosensor, which is characterized in that: a HEMT is used as a transducer, and cholesterol oxidase is fixed on the gate of the transducer, and the gate surface material is gold. The preparation method thereof is also disclosed. The present invention uses a GaN-based high electron mobility transistor (HEMT) structure as a transducer, and fixes cholesterol oxidase on the gold gate of the device to realize the detection of cholesterol. In this way, it not only has the performance of electrochemistry, such as simple testing, simplified structure, real-time reaction, low cost, etc., but also because the intrinsic signal of the HEMT structure is low and it is less affected by other external environments, the sensitivity is greatly improved. It promotes the cholesterol biosensor to improve the accuracy, specificity and stability for cholesterol.

An evaluation apparatus includes an acquiring unit configured to acquire information on a change amount of an absorbance per unit time. The change amount is obtained by a measurement method including causing a peroxidase to act on hydrogen peroxide, which is produced by solubilizing a low-density lipoprotein (LDL) in a specimen with a surfactant that acts on the LDL, and subjecting the resultant to an oxidation reaction with a cholesterol oxidase, to introduce the hydrogen peroxide to a color reaction, followed by measurement of a change of the absorbance with time. The evaluation apparatus also includes an evaluating unit configured to evaluate an amount of an oxidized LDL in the specimen based on the change amount.