Method for extracting and determining activity of glutathione peroxidase in tea trees

A technology of glutathione peroxide and enzyme activity, applied in the field of enzyme detection, can solve the problem that non-enzymatic reactions cannot be deducted, and achieve the effects of reliable measurement results, easy materials and easy control

Inactive Publication Date: 2012-07-18
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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AI Technical Summary

Problems solved by technology

In "Determination of Glutathione Peroxidase Activity in Several Plants", distilled water is used as the non-enzymatic reaction control

Method used

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  • Method for extracting and determining activity of glutathione peroxidase in tea trees
  • Method for extracting and determining activity of glutathione peroxidase in tea trees
  • Method for extracting and determining activity of glutathione peroxidase in tea trees

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Test material: leaves of tea tree variety Longjing 43.

[0026] Prepare the reagents required for the test:

[0027] Sodium azide-phosphate buffer (NaN3-PBS, pH7.0): disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 7.16g, sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O) 3.12g, disodium ethylenediaminetetraacetic acid (EDTA-2Na) 7.44mg, sodium azide (NaNa 3 ) 16.3 mg, dissolved in double distilled water, diluted to 100 ml, and then corrected to pH 7.0 with NaOH solution.

[0028] 10% trichloroacetic acid (TCA) precipitation solution: 10 g of trichloroacetic acid was dissolved in 90 ml of double distilled water.

[0029]0.32mol / L Na 2 HPO 4 : 11.456g disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) Dissolve in distilled water and set the volume to 100ml.

[0030] DTNB chromogenic solution: 20mg DTNB was dissolved in 1% trisodium citrate to 50ml.

[0031] 4mol / L NaOH solution: Dissolve 16g of sodium hydroxide (NaOH) in distilled water to 100ml.

[0032] 1.0mm...

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Abstract

The invention belongs to the field of enzyme detection and discloses a method for extracting and determining the activity of glutathione peroxidase in tea trees. The method adopts leaves of the tea trees as an extracting material and comprises the steps of making a standard curve and extracting crude GSH-Px enzyme liquid, wherein 0.2 mol/L phosphate buffer liquid containing 1 mmol/L of EDTA-2Na and 4% of PPVP and having 6.2 of pH is used for extraction, and the measured activity of GSH-Px is largest. 10% TCA is easy to prepare and has good precipitation effect. The color development reaction reaches the maximum value at 6 min, and 5-thio-2-nitrobenzene anions have maximum light absorption value at 412 nm. The experiment method has the characteristics of easiness in material acquirement, simplicity for operation, easiness in control and reliable determination results.

Description

technical field [0001] The invention belongs to the field of enzyme detection and relates to a method for extracting and measuring glutathione peroxidase activity in tea trees. Background technique [0002] Glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) are important antioxidant enzymes. It is generally believed that glutathione peroxidase (GSH-Px) is a selenium-containing enzyme in mammals. There are many studies on GSH-Px in animals but few studies on GSH-Px in plants. At the same time, with the rapid changes in the world environment, plants are often subjected to various abiotic stresses, and tea trees are no exception. Antioxidant enzymes are an important defense system in plants against abiotic stress. They can remove reactive oxygen species through degradation to resist the oxidative damage induced by abiotic stress, thereby reducing the damage to plants. The determination of the extraction and determination method of GSH-Px in tea tre...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N1/38
Inventor 房婉萍周琳陈暄王玉花张彩丽黎星辉
Owner NANJING AGRICULTURAL UNIVERSITY
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