Method for detecting activity of catalase

A technology of catalase and detection method, which is applied in the detection of catalase activity and the field of enzyme activity detection, and achieves the effect of simple instrument requirements and strong practicability

Inactive Publication Date: 2015-01-21
XIAN MIYI BIOTECH
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AI Technical Summary

Problems solved by technology

[0004] At present, the most widely used determination methods are titration method, ultraviolet rate method, etc. These methods are subject to certain conditions in general operation and application.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Preparation of standard enzyme solution: Dilute the catalase standard product into a series of standard solutions with enzyme activity concentrations for later use.

[0015] Hydrogen peroxide phosphate buffer preparation: take 1mL of 30% hydrogen peroxide, dilute it to 1000mL with 0.01mol / L, pH6.8 phosphate buffer, and set aside.

[0016] Preparation of peroxidase-o-dianisidine solution: Dissolve 0.3 g of peroxidase and 10 g of o-dianisidine in 1000 mL of methanol solution and set aside.

[0017] Take 10mL chlorine peroxide phosphate buffer solution in a 100mL iodine measuring bottle, put it in a 25°C water bath to keep warm, then add 2mL enzyme standard solution, keep warm for 3min accurately, add 2mL0.5mol / sulfuric acid immediately to stop the reaction, add the control reaction solution to the control tube first 2mL of 0.5mol / L sulfuric acid was added to terminate the reaction and then add enzyme solution. Then take 0.1 mL of the above reaction solution plus 2.9 mL o...

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Abstract

The invention relates to a method for detecting activity of catalase, belonging to the field of enzymatic activity detection methods. The method for detecting activity of catalase comprises the following steps: diluting a catalase standard substance into a standard solution with a series of catalase activity concentrations, adding 10mL of chlorine peroxide phosphate buffer solution in a 100mL iodine measuring flask, putting the iodine measuring flask in water bath at 25 DEG C and carrying out heat preservation, then adding 2mL of a standard enzyme solution, accurately carrying out heat preservation for 3 minutes, instantly adding 2mL of 0.5mol/L sulfuric acid to end reaction, adding 2mL of 0.5mol/L sulfuric acid into a contrast tube of a contrast reaction solution to end reaction, and then adding the enzyme solution; then extracting 0.1mL of the reaction solution, adding 2.9mL of a peroxidase-dianisidine solution in a test tube and shaking to be uniform, firstly carrying out zero adjustment by a blank test tube in 436microns by virtue of a spectrophotometer at 25 DEG C, then detecting absorbancy of each reaction solution test tube to obtain a curve about the relation between the standard enzymatic activity and the absorbancy. The test enzyme is processed by the same method, so that the enzymatic activity of the test enzyme can be obtained according to the absorbancy and the standard curve. The detection method has the characteristics of being accurate, high in practicability, simple in instrument requirement and suitable for operation in the conventional condition; the detection method belongs to one reliable method of color rendering methods for detecting the activity of the catalase.

Description

technical field [0001] The invention relates to a method for detecting catalase activity, belonging to the field of enzyme activity detection methods. Background technique [0002] With the development of science and the progress of society, the use of catalase (ECl.11.1.6) is becoming more and more extensive. It is mostly distributed in animal and plant cells, and a large amount of catalase will also be produced in the metabolic process of aerobic microorganisms, and its production and existence are a dynamic process. Animal liver cell metabolism produces a lot of hydrogen peroxide, and this enzyme also has a large amount of savings. Extracting from animal liver is an important way to prepare catalase; bacteria and molds will also release a large amount of catalase in the fermentation process. Extracellular, accumulated in the fermented liquid, through the biochemical preparation method to obtain catalase with higher purity, which can be used in industry and medicine. [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
Inventor 李勇刘文斌
Owner XIAN MIYI BIOTECH
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