59 results about "5'-nucleotidase" patented technology

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

The invention relates to the field of an external diagnostic reagent, and particularly relates to a high-sensitivity kit for detecting 5'-nucleotidase. The kit for detecting 5'-nucleotidase comprises a 5'-nucleotidase R1 reagent, a 5'-nucleotidase R2 reagent and a 5'-nucleotidase calibration product, wherein the 5'-nucleotidase R1 reagent comprises a buffer solution, a stabilizing agent, a preservative agent, an enzyme activator, 4-AAP(4-aminoantipyrine), 18-crown ether-6 and UAO (Uricase), PNP (Purine Nucleoside Phosphorylase), XOD (Xanthine Oxidase) and POD (Peroxidase) reaction enzyme systems; the 5'-nucleotidase R2 reagent comprises a buffer solution, a stabilizing agent, a preservative agent, PO3- and a Trinder developing system; and the 5'-nucleotidase calibration product comprises a buffer solution, a stabilizing agent, a preservative agent and 5'-nucleotidase. The detection kit provided by the invention is enhanced in detection sensitivity by 40% compared with a 'PNP-XOD-POD' enzyme system, and has good application prospects.

The present invention provides ectonucleotidase inhibitors represented by the following formula, including ecto-nucleotide triphosphate diphosphohydrolase (NTPDase) inhibitors and ecto-5′-nucleotidase (ecto-5′-NT) inhibitors, namely nucleotide mimetics as selective NTPDase or ecto-5′-NT inhibitors. It also provides methods for preparations of said compounds. Furthermore provided are pharmaceutical and diagnostic compositions comprising said compounds, and the use of said compounds in a medicament for treating diseases associated with ectonucleotidase activity and / or P1 or P2 receptors.

The present invention relates to a microorganism producing inosine, which is one of purine nucleoside, an important material for 5′-inosinic acid synthesis, and method for producing inosine using the same. More particularly, the present invention relates to a recombinant microorganism of Corynebacterium genus producing inosine at high concentration by inactivating the gene encoding nucleoside hydrolase II and by enhancing the expression of the gene encoding 5′-nucleotidase, which still retains the characteristics of Corynebacterium ammoniagenes CJIP2401 (KCCM-10610).

The invention relates to a method for determining activity of 5′-nucleotidase, in which a biological sample is incubated in a manner and under conditions known per se with a nucleotide pentose monophosphate as substrate, the liberated inorganic phosphate is converted into a colored complex by treating it with ammonium molybdate and a reducing agent, color intensity is measured by a known method, and from the measured value the activity of 5′-nucle-otidase or the amount of inorganic phosphate liberated within unit time, as a figure proportional to the activity of 5′-nucleotidase, is calculated with a known calculation and / or with the aid of a calibration curve. According to the invention 5′-AMP, 5′-CMP, 5′-UMP, 5′-GMP, 5′-IMP and 5′-TMP are used as nucleotide pentose monophosphates, and measurement is performed on all of these six substrates. The invention also relates to a reagent kit for performing the above method.

The invention relates to a 5'-nucleotidase diagnosis reagent box of Enzymatic Recycling Method, and the invention also relates to method principle for detecting the solution of 5'-nucleotidase, constitute and component of reagent, which belongs to the technical field of medical checking measurements. The reagent box in the invention can be dry powder state, and used after dissolution; it also can be formulated to be liquid agent for direct usage. The component of reagent box mainly contains: buffering liquid, alpha- ketoglutaric acid, reduction type coenzyme, Adenosine Deiminase EC 3.5.4.4, glutamate dehydrogenase EC 1.4.1.2, EC 1.4.1.3, EC 1.4.1.4, Glutamate oxidase EC 1.4.3.7, EC 1.4.3.11, peroxidase EC 1.11.1.7, reduction type chromogen assembly, stabilizer and anti-interference agent; and the component can be mixed to form single-reagent reagent box, two-reagent reagent box, and three-reagent reagent box; by series of enzymatic reaction, the colorless reduction type chromogen assembly is oxygenated to form color dye, and the content of dye can be measured by visible light analyzer at wavelength of 400-700nm to reflect the concentration of 5'-nucleotidase directly. Comparing with present technique, the invention can be spread easily, and because the produced dye has higher molar extinction coefficient, the sensitivity is high, and the precision is well.

The invention discloses a xanthine dehydrogenase intercepting body and an application thereof. The protein comprises xanthine dehydrogenase intercepting body small subunit, xanthine dehydrogenase intercepting body intermediate subunit, and xanthine dehydrogenase intercepting body large subunit. The affinity of xanthine dehydrogenase intercepting body is increased by 19% by comparing a wild substrate (xanthine), the conversion number is increased by 115%, the catalysis efficiency is increased by 166%, and the temperature toleration is increased by 11 DEG C. The xanthine dehydrogenase intercepting body is in favor of developing the novel application field by combining with other enzymes, and is suitable for sample detection for detecting xanthine and hypoxanthine, the xanthine dehydrogenase intercepting body can be used for detecting inorganic phosphoric acid by combining with PNP enzyme, can be used for detecting adenosine deaminase by combining with PNP, XOD and POD enzymes and detecting 5'-nucleotidase, and is suitable for industrial application. The xanthine dehydrogenase intercepting body is in favor of increasing enzyme catalysis efficiency and reducing cost, and is in favor of industrial application.

The invention belongs to the technical field of a biosensor, and discloses an enzyme biosensor for detecting inosine monophosphate(IMP) and a preparation method and an application thereof. The lineardetection range of the enzyme biosensor is 0.313-210 microgram / L, and the detection limit is 0.238 microgram / L. The enzyme biosensor is formed by a reference electrode, a counter electrode and a modified electrode obtained by curing an IMP-sensitive substance recognition film on the surface of a working electrode. The substance recognition film is formed by a composite solution a, a double-enzymecomposite solution b formed by a 5'-nucleotidase solution and an xanthine oxidase solution by the volume ratio 1: 1, and a bovine serum albumin solution by the volume ratio of 1: 1: 1, wherein the composite solution a is composed of a MXene-Ti3C2Tx solution (T is selected from OH, O or F), a chitosan solution, a chloroauric acid solution and a chloroplatinic acid solution by the volume ratio of 12:1.2:1:1. The enzyme biosensor is simple, fast and accurate and high in sensitivity, and can be used for quantitative detection of inosine monophosphate(IMP) in food.

The invention discloses a method for constructing an engineering strain for producing flavin mononucleotide, and applications of the engineering strain. According to the method, by using a synthetic biology technology, an intracellular flavinpool component and endometrial secretion are regulated in newly-constructed or existing riboflavin fermenting bacteria, and an inside-cell-membrane-interstitium enzyme catalytic step is novelly added, such that the high-purity flavin mononucleotide is produced through fermentation with microorganisms; and the method specifically comprises: (1) introducinga bifunctional enzyme riboflavin kinase / flavin adenine dinucleotide producing enzyme into riboflavin fermentation bacteria to convert the intracellular flavinpool component from riboflavin to flavin adenine dinucleotide, (2) introducing a flavin adenine dinucleotide internal membrane transporter, and transporting the intracellular flavin adenine dinucleotide into cell membrane interstitium, and (3) introducing 5'-nucleotidase to lyse and convert the flavin adenine dinucleotide in the cell membrane interstitium into the flavin mononucleotide. According to the present invention, under the shaking fermentation condition, the fermentation purity of the flavin mononucleotide (accounting for the total flavin compounds) can be up to 92.4%.

Bertrand(1982) hydrolytic generates trophicardyl from trophicardyl acid by nucleotidase, and gains H2O2 through the reaction of the enzyme1 and enzyme2. Use of H2O2 from Trinder's reaction to combine hydrogen provider3,5-double CL-2-hydroxide benzene sulphur acid and 4- to create a amaranth in the situation of the catalase. By watching the velocity of increase of the degree of the amaranth absorbing light at 510 nms to gain 5' nucleotidase activity. The invention uses the aniline compound ADOS, ADPS, ALPS, TODB, TOOS and TOPS as hydrogen provider for Trinder's reaction and increases the reaction ability. The invention also involves a reagent box used for implementing the method mentioned above.

The invention is directed to a method for improving learning and / or memory (e.g., auditory, visual, somatosensory or motor) in adults and children of an age which is beyond the early critical period for learning, said method comprising inhibiting (i) ecto-5′-nucleotidase (Nt5e, aka CD73) or (ii) A1 adenosine receptor (A1R, aka Adora1) expression or function in the brain. The invention is also directed to a method for treating learning and memory defects and neurological diseases associated with an abnormal auditory, visual, or somatosensory perception by inhibiting Nt5e or A1R expression or function in the brain.

The invention relates to a method for determining the activity of 5'-nucleotidase, further to a 5'-nucleotidase detection kit, and belongs to the technical field of medical test determination. According to the method, 5-inosine monophosphate used as a substrate is converted and dehydrogenated by using purine nucleoside phosphorylase and xanthine dehydrogenase as tool enzymes, oxidized coenzyme isreduced to form a reduced coenzyme, and the enzyme reaction rate is calculated by detecting the generation amount of the reduced coenzyme at a wavelength of 340 nm. Compared to the method in the priorart, the method of the present invention has advantages of less reaction steps, less kinds of raw materials, strong alkaline-phosphatase resistance, stable reagent performance, wide linear range, high sensitivity and good accuracy.

The present invention provides a 5'-nucleotidase detection kit and a preparation method thereof, belongs to the field of in-vitro diagnostic reagents, and solves the low precision and stability problems of a 5'-nucleotidase detection kit in the prior art. The kit comprises a reagent R1 and a reagent R2. The present invention also provides the preparation method of the 5'-nucleotidase detection kit. The R1 reagent contains a compound stabilizer comprising TritonX-305 and AES, the compound stabilizer can improve the stability of the reagent, and also can improve the dispersion of the reagent, reduce the absorbance difference between two times of blank calibration, and improve the kit precision; A90 contained in the R1 can effectively prevent precipitation of magnesium phosphate, expensive 18-crown ether-6 is replaced, the stability of the reagent is improved, reagent blank absorbance change rate is reduced, and the cost is reduced greatly; by addition of glycine and EDTA. 2K into the reagent R2, detection sensitivity and precision of the kit can be improved, and by addition of the glycine, reagent blank absorbance is reduced.

The invention discloses a kit for determination of 5'-nucleotidase and a preparation method thereof, wherein the kit includes double-liquid components of a reagent R1 and a reagent R2 which are independent and includes the components and the corresponding content: the reagent R1 including a Good's buffer liquid, 5'-hypoxanthine nucleotide, xanthine oxidase, purine nucleotide phosphorylase, sodium azide, and a solvent being purified water; and the reagent R2 including peroxidase, 4-aminoantipyrine, 3-hydroxy-2,4,6-tribromobenzoic acid, sodium azide, and a solvent being purified water. The preparation method includes the steps: according to the component content, preparing the reagents; mixing a to-be-tested sample with the reagent R1 and the reagent R2, and carrying out full reaction; determining the absorbance difference value after the reaction with a fully automatic biochemical analyzer; and calculating the concentration of D-3-hydroxybutyrate in the sample according to the absorbance change value. The kit has the advantages of high accuracy and the like.

The invention is about a method of measuring the activation of 5í»-phosphonuclease, and it also concerns the reagent box of 5í»-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, carnine monophosphate, adenosine triphosphate, phosphoenolpyruvate, reduced coenzyme, carnine kinase, pyruvate kinase, lactate dehydrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5í»-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

The present invention provides a 5'-nucleotidase quality control product and belongs to the technical field of quality control products. The quality control product specifically comprises the following components: 10-100 mmol / L of a buffer solution, 0.1-1% of a preservative, 0.1-5% of a protective agent 1, 0.1-5% of a protective agent 2, 0.1-2 mL / L of a surfactant, 1 L of matrix serum and 45-72 U / L of 5'-nucleotidase; the buffer solution is one or more of a TAPSO buffer solution, a potassium dihydrogen phosphate-sodium hydroxide buffer solution, a barbital buffer solution, a MOPS buffer solution, a HEPES buffer solution, a PIPES buffer solution and a phosphate buffer solution; and the protective agent 1 and the protective agent 2 are different and respectively selected from one of glucose,lactose, maltose, maltodextrin and arginine. The 5'-nucleotidase quality control product has an expiration date reaching 24 months.

The invention provides a method for calibrating and measuring the activity of 5'-nucleotidase and a matching kit. In the invention a 5'-nucleotidase dehydrogenase removal method is used to detect the kit for determining the activity of a standard product of the 5'-nucleotidase and then a Trinder' method is applied to quantitatively measure the activity of the 5'-nucleotidase in a biologic sample by taking the activity of the standard product as a reference. The invention has the advantages of solving the defect of the Trinder' method, enhancing the accuracy, reducing the instrument error and reinforcing the data comparability of the normal reference range of the 5'-nucleotidase, is used for the diagnosis and the like of the diseases of primary and transferable liver cancers, biliary tract cancers, pancreatic cancers, biliary tract obstruction, cholangitis, acute and chronic hepatitis, hepatocirrhosis, medicinal hepatic injury and the like by measuring the activity of the 5'-nucleotidase and is suitable for popularization and use in hospitals.

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, nucleoside monophosphate, adenosine diphosphate, an oxaloacetic acid, a ketoglutaric acid, pyruvate carboxylase, isocitrate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, coenzyme, nucleoside monophosphate, glutamine, adenosine diphosphate, glutamine synthetase, glutamate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the increase in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The invention relates to a 5'-nucleotidase from Bailing production bacterium Cordyceps Chinese Hirsutella for synthesizing metabolic pyrimidine from pyrimidine nucleotide, a coding gene and application thereof. The amino acid sequence of the 5'-nucleotidase has more than 90% of homology with sequence disclosed as SEQ ID NO.1; and the coding gene has more than 90% of homology with nucleotide sequence disclosed as SEQ ID NO.2. The invention researches the metabolic pathway of pyrimidine nucleotide synthesized pyrimidine in details in principle. The cloned DNA (deoxyribonucleic acid) comprising the nucleotide sequence provided by the invention can be transformed into engineering bacterium by transduction, conversion and conjugal transfer. By adjusting the expression of the pyrimidine biosynthesized gene, the host pyrimidine is endowed with high expressivity, thereby providing an effective way for enhancing the yield of the pyrimidine and having great application prospects.

The invention is about a method of measuring the activation of 5í»-phosphonuclease, and it also concerns the reagent box of 5í»-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, carnine monophosphate, oxidized coenzyme, carnine nucleotide, ribose1-dehyddrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5í»-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, coenzyme, nucleoside monophosphate, sucrose, sucrose phosphorylase, fructose dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the increase in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, nucleoside monophosphate, an oxaloacetic acid, acetylcoenzyme A, phosphoenolpyruvate carboxykinase, pyruvate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The present invention relates to a microorganism producing inosine, which is one of purine nucleoside, an important material for 5′-inosinic acid synthesis, and method for producing inosine using the same. More particularly, the present invention relates to a recombinant microorganism of Corynebacterium genus producing inosine at high concentration by inactivating the gene encoding nucleoside hydrolase II and by enhancing the expression of the gene encoding 5′-nucleotidase, which still retains the characteristics of Corynebacterium ammoniagenes CJIP2401 (KCCM-10610).

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, nucleoside monophosphate, adenosine diphosphate, an oxaloacetic acid, phosphoenolpyruvate carboxykinase, formaldehyde deoxygenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The invention is about a method of measuring the activation of 5í»-phosphonuclease, and it also concerns the reagent box of 5í»-phosphonucleasediagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, uridine monophosphate, reduced coenzyme, uridine phosphorylase, dihydrothymine dehyddrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get theactivation of 5í»-phosphonuclease. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

The invention relates to a kit for diagnosing 5'-nucleotidase by utilizing the technologies of the enzymic colorimetry and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for measuring the active concentration of 5'-nucleotidase, and belongs to the technical field of medical inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, nucleoside monophosphate, adenosine diphosphate, an oxaloacetic acid, ammonium chloride, pyruvate carboxylase, alanine dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the velocity of the decrease in absorbance at 340 nm of the dominant wavelength is detected, thereby measuring the active concentration of 5'-nucleotidase.

The invention relates to a kit for diagnosing / mensurating inorganic phosphorus by utilizing the technologies of the enzymic colorimetric method and the enzyme linked immunosorbent assay. The invention further relates to a method, a principle and the composition and the components of a reagent for mensurating the concentration of the inorganic phosphorus, and belongs to the technology field of medical / food / environmental inspection and measurement. The main components of the kit include a buffer solution, coenzyme, inosine, 5-nucleotidase, inosine monophosphate dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet / visible light analyzer, and the degree / velocity of the increase in absorbance at 340nm of the dominant wavelength is detected, thereby mensurating the concentration of the inorganic phosphorus.