Method for determining activity of soil arginine desaminase

A kind of technology of arginine deaminase and soil, applied in the direction of biochemical equipment and methods, determination/inspection of microorganisms, measurement of color/spectral characteristics, etc., can solve the problem that the determination of arginine deaminase activity is not accurate Reliable and simple determination methods, limited application research and other issues, to achieve stable and reliable analysis results, reduce dependence, and stabilize blue compounds

Inactive Publication Date: 2011-06-29
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

So far, there is no accurate, reliable and simple method for the determination of arginine deaminase activity, which greatly limits its application research

Method used

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  • Method for determining activity of soil arginine desaminase

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Embodiment

[0048] The brown soil used in this example was collected from the Shenyang Ecological Experimental Station of the Chinese Academy of Sciences, and the cinnamon soil was collected from the experimental field of the Chaoyang Soil and Fertilizer Station in Liaoning. Both soils were cultivated at 25 ° C for more than 24 hours, and the moisture content was about 15%. They were detected by the above method Arginine deaminase activity in two soils.

[0049] The specific analysis steps for each soil are as follows:

[0050] 1) Weigh 4 parts of 5g fresh soil through a 2mm sieve into a 150mL conical flask, add 2mL of 11.5mol / L arginine substrate solution to the conical flask, mix the soil and substrate solution evenly, and seal the plug rubber stoppers; two of the flasks were incubated in a 37°C incubator at a constant temperature for 3h (sample), and the remaining flasks were incubated at a temperature of -20°C for 3h (control); Next to thaw. At the same time, it was used as reagent ...

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Abstract

The invention relates to a method for determining the activity of soil arginine desaminase, which comprises the following steps: 1) weighting n fresh soil or precultured soil samples, placing in n conical flasks having plugs (wherein n is more than or equal to 4), respectively adding arginine substrate solution into each conical flask, uniformly mixing the arginine substrate solution with the soil, blocking each flask with a rubber plug, and then culturing for 3 hours at constant temperature; 2) after the culturing process is finished, adding a certain volume of potassium chloride solution into the flasks, oscillating, and filtering; 3) determining the quantity of generated ammonium ions by using the indophenol blue colorimetry; and 4) calculating the activity of arginine desaminase. The method for determining the activity of soil arginine desaminase has the following advantages: 1) compared with the traditional biochemical determining method, the method provided by the invention simplifies the operation steps; 2) the dependence on high equipment demand is reduced, and a common spectroscopic instrument can meet the demand; 3) the accuracy is high, and the operation can be easily performed; and 4) the result is stable and reliable, and the repeatability is good.

Description

technical field [0001] The invention relates to the measurement of soil arginine deaminase activity, in particular to a new method for detecting soil arginine deaminase activity. Background technique [0002] Arginine deaminase in soil is a kind of arginine lyase, which is involved in the transformation cycle of arginine. Arginine is a semi-essential or conditional essential basic amino acid, which is hydrolyzed under the action of arginine deaminase to generate ornithine and urea, and the formed urea is released in the soil for plant absorption and utilization. It can be seen that, Arginine is an important nitrogen storehouse, it can use the smallest carbon to combine the most nitrogen, and it is the amino acid with the highest N / C ratio among the 18 essential amino acids. [0003] Therefore, the strength of arginine deaminase activity in soil affects the nitrogen metabolism process of soil urea, and then affects the nitrogen use efficiency. The activity of arginine deami...

Claims

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Application Information

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IPC IPC(8): C12Q1/527G01N21/31G01N21/78
Inventor 隽英华陈利军武志杰
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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