39 results about "Oxalate oxidase" patented technology

The invention discloses a reagent, a kit and a method for detecting the content of oxalic acid in urine and blood and relates to the field of scientific research and detection for medical oxalic acid. The reagent comprises a buffer solution of which the pH value is 4.0-7.5, Amplex red fluorescent dye of which the concentration is 1-50[mu]mol/L, horse radish peroxidase (HRP) of which the concentration is 0.1-2U/ml and oxalate oxidase of which the concentration is 0.01-1.0U/ML. The method comprises the following steps of collecting and purifying a sample to be detected to obtain a purified liquid to be detected; dividing the liquid to be detected into two parts; preparing the liquid to be detected into a calibrated liquid to be detected; dividing the liquid to be detected of each part into two groups, namely a determining group and a comparing group; adding an oxalic acid determining reagent in the determining group to obtain a fluorescent group to be detected; adding an oxalic acid background reagent in the comparing group to obtain a determined fluorescent value after comparing the groups to be detected, and obtaining the detection limit of the oxalic acid, namely 1.5-3mol/L. The reagent, the kit and the method disclosed by the invention, is used for scientific research and clinical medicine, and high throughout detection can be realized; the method is high in sensitivity and better in reliability.

The invention discloses a reagent, a kit and a piece of test paper for determining the concentration of oxalic acid, and a preparation method for the test paper, and relates to the field of early diagnosis and prevention of kidney stone. The reagent comprises oxalate oxidase with the concentration of 10 to 2,000 U/L and the optimal pH value of 4.0 to 7.5, horseradish peroxidase with the concentration of 100 to 2,000 U/L, and a reactant with the concentration of 10 to 200 mmol/L, wherein the reactant is an oxidant or a developing agent; the oxidant is benzoquinone or potassium ferricyanide; the developing agent is 3-methyl-benzothiazolinone hydrazone hydrochloride or sodium 3,5-dichloro-2-hydroxybenzenesulfonate. Electrochemical test paper and light reflecting test paper disclosed by the invention are convenient to use, easy to operate, short in determination time, high in accuracy and anti-jamming capability, and long in storage time, and can be used in a hospital, a clinic or a family.

A process for culturing the antifungal transgenic rape with dual-gene specific expression includes using RCR or restriction endonuclease to separate the amino acid coding sequences of chitinase gene and oxalate oxidase inplant and the promoter sequence specifically expressing the senile leaf, configuring the recombinant dual-gene expression carrier, introducing the exogenous chitinase gene and oxalate oxidase gene to rape receptor to obtain transgenic plant, and molecular auxiliary selection while conventional culturing.

The invention is a method for extracting oxalate oxidase from wheat bran and an enzyme preparing technique, washing the wheat bran by water, then homogenizing by buffer solution, and centrifuging and collecting deposit; using the same buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting the deposit; successively suspending the deposit with buffer solution containing 1mol/L NaCl, preserving the heat during water bath for 1-1.5 h and then centrifuging and collecting deposit; using buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting deposit; adding butter solution in the deposit, making boiling water bath for 15-20 minutes and centrifuging and collecting deposit, using buffer solution or distilled water to drop-wash the deposit until the eluent is clear and then collecting deposit; then suspending the deposit by water solution containing pepsin, preserving the heat during water bath for 1-1.5 h and then centrifuging and collecting deposit, using buffer solution or distilled water to drop-wash the deposit until the eluent is clear; and finally using vacuum condense-drying system to eliminate the water in the deposit, thus obtaining the oxalate oxidase.

The invention relates to a simple method for determining oxalic acid concentration. The method comprises the following steps: (1) reacting synthetic 3NO ligand with manganese perchlorate to prepare a manganese complex; (2) reacting the prepared metal manganese complex with standard hydrogen peroxide sample with concentration being 1-400mum, determining absorbance of the reaction product at the wavelength being 450 nm, and making a standard curve of absorbance of the standard hydrogen peroxide sample at the wavelength being 450 nm; (3) reacting oxalic acid sample having unknown concentration with excessive oxalate oxidase, and decomposing the oxalic acid into equal amount of hydrogen peroxide; and (4) reacting the product of the step (3) with the metal complex prepared in the step (1), determining the absorbance of the product at the wavelength being 450 nm, and comparing with the standard curve of the absorbance of the standard hydrogen peroxide sample at the wavelength being 450 nm to obtain the concentration of the oxalic acid in the sample. The method has the advantages of simple step, high flexibility, low cost and short time.

Methods for identifying one or more amino acid substitutions in an oxalate oxidase (OXOX) variant polypeptide that confer maintained or increased OXOX activity are described herein. Methods and compositions for increasing a plant's resistance to a pathogen using the modified OXOX variant polypeptides are provided. Transformed plants, plant cell, tissues, seed, and expression vectors are also provided.

The invention relates to an electrochemical biosensor for detecting oxalic acid concentration, and preparation and application thereof. Oxalic acid oxidase is fixed on the surface of part of reduction-oxidation grapheme, a dispensing method is adopted to enable immobilized oxalic acid oxidase to be decorated on the surface of a glassy carbon electrode, and the electrochemical biosensor based on decoration of oxalic acid oxidase and part of reduction-oxidation grapheme is formed. The glassy carbon electrode serves as a working electrode, and a difference volt-ampere pulse method is adopted to detect the oxalic acid concentration. The biosensor has good electro-catalysis activity on oxalic acid. Along with increase of the immobilization amount, the electro-catalysis activity is reinforced. The detection range of the electrode is wide, and the electrode has low detection lower limit and responds rapidly.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme multiplication method, an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxalic acid, acetyl coenzyme A, ammonium chloride, oxalate oxidase, pyruvate dehydrogenase, alanine dehydrogenase, and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention discloses an Escherichia coli expression system for producing oxalate oxidase, a production method of oxalate oxidase and application thereof. The recombinant expression plasmid in the E. coli expression system comprises: an oxalate oxidase gene, a chaperone gene, and a gene that promotes to pump manganese ions into E. coli cell-associated protein. By screening and testing the genesof different manganese ion-related proteins and their copy number, the type of a promoter for controlling expression of the molecular chaperone and the type of a culture medium used in the productionprocess of oxalate oxidase, an optimized expression combination of oxalate oxidase is obtained, and the expression activity of oxalate oxidase in E. coli exceeds 80 U/mL. The specific activity of theoxalate oxidase preliminarily purified by a supernatant of a cell disrupted liquid is greater than 20 U/mg, and the molecular weight of the monomer is about 25 kDa. The technical scheme of the invention has the advantages of simple production and purification process, high expression level and specific activity, easy industrial amplification and low cost, and is advantageous for industrial production and application of such enzymes.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme multiplication method, an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxalic acid acetyl coenzyme A, oxalate oxidase, pyruvate dehydrogenase, lactate dehydrogenase, and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention relates to a kit for diagnosing/mensurating manganese ions 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 manganese ions, and belongs to the technology field of medical/food/environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, oxalic acid, pyruvic acid, oxalate oxidase, malate 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 340nm of the dominant wavelength is detected, thereby mensurating the concentration of the manganese ions.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxalic acid, acetic acid, ferrocytochrome b1, oxalate oxidase, pyruvate dehydrogenase, lactate dehydrogenase, and a stabilizer. The method comprises the following steps: mixing samples and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxalic acid, acetic acid, ferrocytochrome b1, oxalate oxidase, pyruvate dehydrogenase, malic dehydrogenase, and a stabilizer. The method comprises the following steps: mixing samples and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention relates to an application of a photorespiratory branch protein in regulation and control of plant traits. Specifically, the invention provides a reagent combination and a fusion protein, and the reagent combination or the fusion protein comprises glycollic acid oxidase or coding nucleic acid thereof and/or oxalate oxidase or coding nucleic acid thereof and/or catalase or coding nucleic acid thereof. The agronomic traits of plants can be remarkably regulated and controlled by introducing the reagent combination or the fusion protein disclosed by the invention into plant cells.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, coenzyme, oxalic acid, oxalate oxidase, NAD(P)H oxidase and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the ascending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention relates to a manganese ion diagnosis/determination kit using the technology of an enzyme multiplication method, an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxalic acid, acetyl coenzyme A, oxalate oxidase, pyruvate dehydrogenase, malic dehydrogenase, and a stabilizer. The method comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.

The invention discloses a urine oxalic acid content detection system and detection method. The detection system comprises a pretreatment kit and a detection kit; the pretreatment kit comprises activated carbon and a sample treatment solution, wherein the detection kit comprises a reagent R1 and a reagent R2; the R1 reagent is prepared from the following components: citric acid, DMAB and MBTH; the reagent R2 comprises the following components: a buffer solution with the pH value of 5.6, oxalate oxidase and peroxidase; the concentration of the buffer solution in the R2 reagent is 0.1 M, the concentration of the oxalate oxidase in the R2 reagent is greater than 50 U/L, and the concentration of the peroxidase in the R2 reagent is greater than 5000 U/L; and the R2 reagent is in a dry powder state and is redissolved by water when being used. The method has a high detection limit, can be used for detecting the content of oxalic acid in urine, has high detection sensitivity, and can eliminate interference of substances such as vitamin C in urine, so that the detection result is more accurate.

The invention relates to a manganese diagnosis/determination reagent kit which adopts the techniques of enzymatic colorimetric method and enzyme couple reaction, belonging to the technical field of medicine/food/environment test and determination. The invention also relates to the method principle of determining the manganese concentration, and the reagent composition and components. The component of the reagent kit mainly comprises buffer solution, oxalic acid, oxalate oxidase, peroxidase, reduced chromogen combination, and stabilizer. The method for determining the manganese concentration is as follow: oxidizing the colorless reduced chromogen combination into colored dye through a series of enzymatic reactions; determining the dye content at the 400 to 700 nm wavelength with a visible light analyzer; finally determining the manganese concentration directly.