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Analysis method for detecting activity of beta-D-glucosidase in soil

A technology of glucosidase and analysis method, which is applied in the analysis field of detecting β-D-glucosidase activity in soil, and can solve problems such as the influence of result accuracy

Pending Publication Date: 2019-04-02
JILIN ACAD OF AGRI SCI
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Problems solved by technology

The widely used fluorescence detection technology of enzyme activity nowadays excludes the background value of soil and substrate only through the no-soil control and no-substrate control. For fluorescence detection, which is a very sensitive detection method, any background interference will affect the accuracy of the results. sex has considerable influence

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  • Analysis method for detecting activity of beta-D-glucosidase in soil
  • Analysis method for detecting activity of beta-D-glucosidase in soil
  • Analysis method for detecting activity of beta-D-glucosidase in soil

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Embodiment Construction

[0021] A specific embodiment of the present invention will be described in detail below, but it should be understood that the protection scope of the present invention is not limited by the specific embodiment.

[0022] Next, the β-D-glucosidase activity of the three treatments of corn continuous cropping (A), soybean rice rotation (B) and soybean rice rice rotation (C) were measured. The soils of the three different cropping systems were all over Fresh soil after 2mm sieve.

[0023] The specific analysis steps for each soil are as follows:

[0024] 1. Preparation and determination of standard song: prepare standard solution, weigh 1.762 mg of 4-methylumbelliferone and 30 μl dimethyl sulfoxide to dissolve, and then adjust the volume to 1000 ml to obtain a 10 μM standard solution, and mix 10 μM of Dilute the standard solution with distilled water into standard solutions with concentrations of 0, 0.025, 0.05, 0.125, 0.25, and 0.5 μM to prepare the standard solution, pipette 50 ...

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Abstract

The invention discloses an analysis method for detecting activity of beta-D-glucosidase in soil, and belongs to the technical field of enzyme activity detection. According to the method, the enzyme activity is equal to [(RS-BU)*(SA-SC) / (QS-SC)]-(NC-BU)*B*2*300 / (0.3*W*200), wherein SC is a fluorescence intensity value measured by supernatant and 50 ul of acetic acid buffer solution, QS is a fluorescence intensity value measured by supernatant and 50 ul of master standard solution, SA is a fluorescence intensity value measured by supernatant and 50 uL of 4-methylumbelliferone-beta-D-glucosidasesubstrate solution, BU is a fluorescence intensity value measured by 200 ul of acetic acid buffer solution and 50 ul of acetic acid buffer solution, RS is a fluorescence intensity value measured by 200 ul of acetic acid buffer solution and 50 ul of standard substance solution, and NC is a fluorescence intensity value measured by 200 ul of acetic acid solution and 50 ul of 4-methylumbelliferone-beta-D-glucosidase substrate solution. The method is capable of decreasing the background values, eliminating the background values interacted by samples and fluorophores, eliminating the background values, for standard substances, of buffer solution, improving the correctness of measurement results, and making the sample and drug dosages less.

Description

technical field [0001] The invention relates to the technical field of enzyme activity detection, in particular to an analysis method for detecting β-D-glucosidase activity in soil. Background technique [0002] Soil extracellular enzymes play an important role in the ecosystem, and their activity reflects the metabolism of soil microorganisms, which is extremely important for maintaining soil fertility and plant productivity. β-D-glucosidase, a more accurate analytical method for detecting β-D-glucosidase activity in soil, is currently the most widely studied soil enzyme, deeply involved in the transformation of soil organic matter, mainly composed of fiber The secretion of microorganisms that use carbon as substrate, hydrolyze cellobiose and other water-soluble cellodextrins to produce glucose, which can be used by microorganisms for their own growth, and can indicate the intensity of heterotrophic respiration using organic or inorganic carbon in soil as substrates. play ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 孟凡钢张伟闫晓艳赵婧邱强张鸣浩于德斌饶德民闫日红丛博韬颜秀娟李明姝王新风
Owner JILIN ACAD OF AGRI SCI
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