Enzyme catalysis detection method of selenium monosaccharide

A monosaccharide and monooxygenase technology, which is applied in the direction of material analysis by chemical reaction of materials, material analysis by observing the influence on chemical indicators, measurement devices, etc., to save labor costs, simplify the identification process, save Effects of hardware devices

Active Publication Date: 2020-05-26
迈科若(苏州)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, how to use the above reaction to detect selenium monosaccharide GalNAc1SeMe, people have not yet developed a detection method with simple process and high detection efficiency.

Method used

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  • Enzyme catalysis detection method of selenium monosaccharide
  • Enzyme catalysis detection method of selenium monosaccharide
  • Enzyme catalysis detection method of selenium monosaccharide

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Embodiment

[0036]In order to verify the ability of cytochrome P450 monooxygenase, ferredoxin, ferredoxin reductase and reduced tetramethylbenzidine to detect selenomonosaccharide GalNAc1SeMe, the selenomonosaccharide GalNAc1SeMe solution was used as the substrate to Cytochrome P450 monooxygenase, ferredoxin, ferredoxin reductase and reduced tetramethylbenzidine were added to the solution, and if the solution changed color, it indicated that selenium monosaccharide could be detected.

[0037] The preparation method of selenium monosaccharide GalNAcSeMe solution is as follows:

[0038] The powder sample standard of GalNAc1SeMe was vortex-dissolved with 50 mM pH 7.4 sodium phosphate buffer solution, and prepared into a 100 μM stock solution, which was left to mutate overnight in a refrigerator at 4°C to obtain a substrate solution.

[0039] Cytochrome P450 monooxygenase is recombinantly expressed by Escherichia coli (E.coli for short), and is derived from the bacteria Formosa agariphila. Th...

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Abstract

The invention relates to an application of cytochrome P450 monooxygenase (CYP236A20), ferredoxin (FoX), ferredoxin reductase (FoR) and tetramethyl benzidine (TMBred) in detection of selenium monosaccharide or preparation of a selenium monosaccharide detection reagent. The detected selenium monosaccharide is 1beta-methylseleno-2-N-acetyl-D-galactosamine (GalNAc1SeMe), and the detected selenium monosaccharide is a metabolic excretion in urine of a human body. The invention further provides a new detection method of the selenium monosaccharide, namely, CYP236A20 is used as a catalyst and a receptor of a substrate, FoX and FoR are used as electron transfer media, and reduced TMBred is used as a proton donor and a color developing agent; the ligand is removed by utilizing the recognition of enzyme and a methyl selenium functional group in a to-be-detected object, and the color of the solution is changed from colorless transparency to bluish violet along with tetramethyl benzidine oxidation(TMBox), so that the sensitive indication of the selenium monosaccharide is realized.

Description

technical field [0001] The invention relates to a detection method of selenium monosaccharide, in particular to an enzyme-catalyzed detection method of selenium monosaccharide. Background technique [0002] Selenium is an essential trace mineral for the human body, and "Keshan disease" caused by selenium deficiency occurs commonly in remote areas in northern my country and central and western China; modern clinical medicine has further confirmed that the content of selenium in the human body is related to many disease-related molecules (such as cardiovascular disease). The metabolic expression of the marker-homocysteine) is closely related. Therefore, it is extremely necessary to establish biochemical indicators to evaluate the nutritional level of selenium. However, according to the known selenium synthesis and metabolism, the structure of selenoprotein P, which is the selenium pool in human plasma, is poorly conserved and has an expression threshold, while the expression o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/78G01N21/3103
Inventor 邓盛元王丽娟黄洁吴希郭睿李臻怡宋美燕王健郭仁妹
Owner 迈科若(苏州)医疗科技有限公司
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