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Method for detecting activity of plant superoxide dismutase, catalase and peroxidase

A technology of catalase and peroxidase, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, can solve the problems of difficult batch operation, large sample demand, time-consuming and labor-intensive, etc., and achieve high extraction efficiency , Grinding speed is fast, the effect of reducing dosage

Inactive Publication Date: 2010-10-27
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] The main purpose of the present invention is to detect SOD, CAT and POD three kinds of enzyme activities in the above-mentioned prior art, adopt the ice-bath grinding method step loaded down with trivial details, time-consuming and laborious, batch operation difficulty, and need the phosphate buffer of different concentration and pH value Extraction and detection of the three enzymes in different liquids, the need for more samples, the greater workload, etc., provides a method for detecting the activity of plant superoxide dismutase, catalase and peroxidase, using a The three enzymes of SOD, CAT and POD in plants can be extracted at the same time with the buffer solution, which can simplify the operation, save time and effort, and reduce the amount of samples and reagents, etc.

Method used

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  • Method for detecting activity of plant superoxide dismutase, catalase and peroxidase
  • Method for detecting activity of plant superoxide dismutase, catalase and peroxidase
  • Method for detecting activity of plant superoxide dismutase, catalase and peroxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] This example compares the effects of liquid nitrogen grinding method and ice bath grinding method on the extraction efficiency of SOD, CAT and POD enzymes.

[0053] Taking the detection of SOD, CAT and POD enzyme activities in potato leaves as an example, fresh potato leaves were chopped and mixed, and then ground using ice bath grinding method and liquid nitrogen grinding method to extract the phosphate buffer and phosphate buffer solution of the above three enzymes. Enzyme activity determination was carried out in accordance with traditional methods, and the experiment was set up for 6 repetitions. The specific operations are as follows:

[0054] (1) Sample grinding and enzyme extraction:

[0055] A. Ice bath grinding method: Take 1g leaves in a pre-cooled mortar, add pre-cooled 0.05mol / L, pH 7.8 phosphate buffer, grind the homogenate on ice, dilute the volume to 10 mL, and extract for 30 min. Centrifuge at 12000g for 2 minutes at 4°C, and take the supernatant that is the S...

Embodiment 2

[0062] This example is to compare the effect of a method of simultaneously extracting the above three enzymes with a phosphate buffer solution of the present invention and a traditional method of extracting the above three enzymes with three traditional phosphate buffer solutions on enzyme activity.

[0063] Taking the detection of CAT, POD and SOD enzyme activities in safflower leaves as an example, the fresh safflower leaves are chopped and mixed, and three traditional phosphate buffers are used to extract the above three enzymes and a phosphate of the present invention. The buffer solution extracts the above three enzymes at the same time, and the enzyme activity determination is carried out according to the traditional method. The experiment is set up for 6 repetitions. The specific operation is as follows:

[0064] (1) Sample grinding and enzyme extraction:

[0065] A. The three kinds of traditional phosphate buffer solutions are used to extract the above three enzymes: Take 1g...

Embodiment 3

[0072] This example uses the method of the present invention to detect the effect of low-temperature culture on the enzyme activities of SOD, CAT and POD in Houttuynia cordata leaves. The test is set to be repeated 6 times, and the specific operations are as follows:

[0073] (1) Sample grinding and enzyme extraction:

[0074] Take 1g of Houttuynia cordata leaves cultured (cultivated) at 10°C (low temperature) and 25°C (control) respectively, place them in a pre-cooled mortar, pour into liquid nitrogen and grind, and then add 10 mL of 0.2mol / L , PH 7.0 phosphate buffer solution for 30 min, centrifugation at 4 ℃, 12000g for 2 min, take the supernatant is the mixed enzyme solution of the above three enzymes;

[0075] (2) Determination of enzyme activity:

[0076] The SOD enzyme activity was determined by the nitrotetrazolium light blue method, the CAT enzyme activity was determined by the ultraviolet absorption method, and the POD enzyme activity was determined by the guaiacol method. ...

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Abstract

The invention discloses a method detecting activity of plant superoxide dismutase, catalase and peroxidase, which comprises the steps of grinding a sample by using a liquid nitrogen method, extracting three enzymes simultaneously by adopting one buffer solution, detecting enzyme activity and the like. In the method, SOD, CAT and POD in plants can be simultaneously extracted by adopting one buffer solution, so that the method fulfills the aim of simply and quickly detecting the enzyme activity of the SOD, CAT and POD, simplifies operation, saves time and labor, reduces the using amount of samples and reagent and the like, and contributes to the realization of batch operation, and the detection result is similar to that of the conventional method. Furthermore, the sample is ground by adopting a liquid nitrogen grinding method before enzyme solution is extracted, so that the method has the advantages of high grinding speed, good cell crushing effect and high extraction efficiency.

Description

technical field [0001] The invention belongs to the technical field of detection methods for plant superoxide dismutase, catalase and peroxidase activity, and in particular relates to a detection method for plant superoxide dismutase, catalase and peroxidase activity. Background technique [0002] During the physiological metabolism of plants, part of the electron escapes to produce high-energy reactive oxygen species with toxic effects, including O 2 - 、H 2 o 2 , singlet oxygen, hydroxyl radicals, etc. Chloroplasts, mitochondria, and plasma membranes of plant cells are the main places where reactive oxygen species are produced. The accumulation of these reactive oxygen species can subject plant cells to oxygen stress. In order to reduce the damage of reactive oxygen species to cells and maintain the dynamic balance of reactive oxygen species in the body, plants have gradually formed a set of antioxidant enzymatic systems during evolution, including superoxide dismutase...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/30C12Q1/28
Inventor 吴卫徐应文刘正琼邵金凤赵丹杨文婷
Owner SICHUAN AGRI UNIV
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