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Testing method of enzyme activity of cysteine sulfinate decarboxylase in marine organism body

A cysteine ​​sulfinate decarboxylase and marine organism technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, measuring devices, etc., can solve the problems of unsuitability for cysteine ​​sulfinate decarboxylase and high requirements, Achieve the effect of low detection cost and universal applicability

Inactive Publication Date: 2017-10-17
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method has specific applicability and is suitable for the determination of the activity of deiodinase, and the determination of the radioactive intensity of iodide ion has high requirements on the instrument, so it is not suitable for the determination of the enzyme activity of cysteine ​​sulfinic acid decarboxylase

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  • Testing method of enzyme activity of cysteine sulfinate decarboxylase in marine organism body
  • Testing method of enzyme activity of cysteine sulfinate decarboxylase in marine organism body

Examples

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Embodiment 1

[0033] A method for measuring the enzyme activity of cysteine ​​sulfinate decarboxylase in marine organisms. The solution containing cysteine ​​sulfinate decarboxylase is isolated from the tissue samples of marine organisms, and the added sulfinic acid decarboxylase is added under suitable reaction conditions. Amylalanine was converted into taurine, the content of the generated taurine was determined, and then the enzyme activity of cysteine ​​sulfinic acid decarboxylase was calculated.

[0034] Determining the content of taurine produced preferably comprises the steps of first adding a suitable derivatizing reagent to the taurine-containing solution, and then measuring the concentration of the derivative by high performance liquid chromatography. Wherein the derivatization reagent is preferably prepared through the following process: dissolve 0.01g of o-phthalaldehyde in 1mL of methanol, then add 0.01mL of ethanethiol and mix, then dilute to 0.4mol / L sodium borate buffer solut...

Embodiment 2

[0048] A method for measuring the enzyme activity of cysteine ​​sulfinate decarboxylase in marine organisms. The solution containing cysteine ​​sulfinate decarboxylase is isolated from the tissue samples of marine organisms, and the added sulfinic acid decarboxylase is added under suitable reaction conditions. Amylalanine was converted into taurine, the content of the generated taurine was determined, and then the enzyme activity of cysteine ​​sulfinic acid decarboxylase was calculated.

[0049] Determining the content of taurine produced preferably comprises the steps of first adding a suitable derivatizing reagent to the taurine-containing solution, and then measuring the concentration of the derivative by high performance liquid chromatography. Wherein the derivatization reagent is preferably prepared through the following process: dissolve 0.01g of o-phthalaldehyde in 1mL of methanol, then add 0.01mL of ethanethiol and mix, then dilute to 0.4mol / L sodium borate buffer solut...

Embodiment 3

[0063] A method for measuring the enzyme activity of cysteine ​​sulfinate decarboxylase in marine organisms. The solution containing cysteine ​​sulfinate decarboxylase is isolated from the tissue samples of marine organisms, and the added sulfinic acid decarboxylase is added under suitable reaction conditions. Amylalanine was converted into taurine, the content of the generated taurine was determined, and then the enzyme activity of cysteine ​​sulfinic acid decarboxylase was calculated.

[0064] Determining the content of taurine produced preferably comprises the steps of first adding a suitable derivatizing reagent to the taurine-containing solution, and then measuring the concentration of the derivative by high performance liquid chromatography. Wherein the derivatization reagent is preferably prepared through the following process: dissolve 0.01g of o-phthalaldehyde in 1mL of methanol, then add 0.01mL of ethanethiol and mix, then dilute to 0.4mol / L sodium borate buffer solut...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a testing method of enzyme activity of cysteine sulfinate decarboxylase in a marine organism body. The method includes steps of separating solution containing cysteine sulfinate decarboxylase from an organization sample of the marine organism body; transforming the added cysteic acid to taurine under a proper reaction condition; adding proper derivatization reagent in the reaction fluid to test the amount of the generated taurine, and further calculating the enzyme activity of the cysteine sulfinate decarboxylase, wherein the derivatization reagent is acquired by dissolving phthalic dicarboxaldehyde in methanol, adding ethanethiol to mix and then fixing capacity by sodium borate buffer fluid; the enzyme activity of cysteine sulfinate decarboxylase in the marine organism body can be accurately, simply and safely tested; the detection cost is low; meanwhile, the method also can be applied to test enzyme activity of cysteine sulfinate decarboxylasein other animal bodies; therefore, the method has universal applicability.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a method for measuring the enzyme activity of cysteine ​​sulfinic acid decarboxylase in marine organisms. Background technique [0002] Cysteine ​​sulfinic acid decarboxylase is an amino acid decarboxylase enzyme used to catalyze the decarboxylation of cysteine ​​sulfinic acid to produce aminoethylsulfinic acid and carbon dioxide. Aminoethylsulfinic acid is dehydrogenated to generate taurine. The enzyme can be seen in the liver, spleen, kidney, small intestine wall, etc. of animals, with pyridoxal phosphate as the coenzyme. Taurine is a sulfur-containing non-protein amino acid that exists in a free state in animals and is an active substance that regulates the normal physiological activities of the body. It has a wide range of physiological functions and can enhance the antioxidant capacity of cells, protect cardiomyocytes, improve Nerve conduction and visual functi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/527G01N30/02G01N30/06
CPCC12Q1/527G01N30/02G01N30/06
Inventor 柳小英毛丽沙鲍敏洁金火喜
Owner ZHEJIANG OCEAN UNIV