Analyzing method for detecting activity of soil xylanase

A technology of cellulase and analysis method, applied in the direction of material stimulation analysis, biochemical equipment and methods, measurement/inspection of microorganisms, etc., which can solve the problems of increasing the risk of operators, instability of chromogenic substances, poor measurement reproducibility, etc. problems, to achieve stable and reliable analysis results, fewer reagent varieties, and good reproducibility

Inactive Publication Date: 2009-11-25
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

The steps of this method are cumbersome, and the chromogenic substance is unstable, and the measurement reproducibility is poor. Then Schinner and van Mersi improved the assay method in the monograph Methods in Soil Biology written by Schinner and van Mersi in 1996, and the chromogenic substance is relatively stable. Reproducibility is better, but similarly, its determination process is more complicated, takes a long time, and the required reagent potassium cyanide in the color development process is highly toxic reagent, is difficult to obtain, and has increased in the determination process danger to the operator

Method used

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  • Analyzing method for detecting activity of soil xylanase
  • Analyzing method for detecting activity of soil xylanase

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Experimental program
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Embodiment 1

[0037] The soil used in this example was collected from Nianyu Farm (31°37’N, 120°28’E) in Anzhen Town, Wuxi City, Jiangsu Province, a rice-wheat rotation FACE (Free-air Carbon Elevation open CO 2 Concentration increased) system platform, the platform has 3 FACE test circles and 5 control circles, and the FACE test circle keeps CO 2 The concentration is 200μmol·mol higher than the control circle -1 . The soil type is hydroponic man-made soil (commonly known as yellow soil). Two treatments are CO 2 Increased concentration and control, CO 2 The treatment with increased concentration was repeated 3 times, and the control was repeated 5 times. Samples were taken at the mature stage of wheat growth and the soil cellulase activity was detected by the above method.

[0038] The specific analysis steps for each soil are as follows:

[0039] 1) Weigh 3 parts of 3g air-dried soil samples passed through a 2mm sieve into rough test tubes, and add 40ml of 0.5mol L to each test tube -...

Embodiment 2

[0052] The three kinds of black soils used in this example were all collected in Hailun County, Suihua City, Heilongjiang Province: the soil in treatment 1 was continuous cropping of soybeans, the soil in treatment 2 was continuous cropping of rice, and the soil in treatment 3 was continuous cropping of corn. The soils of three different cropping systems were tested after passing through a 2mm sieve. Concrete steps are with embodiment 1.

[0053] The test results are as follows:

[0054]

[0055] The data in the table also shows the stability and high precision of the analysis results.

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Abstract

The invention relates to an analyzing method for detecting activity of soil xylanase, which comprises the following steps: firstly, weighting n sieved air dry soil samples into n thick test tubes, adding acetic acid buffer solution into each test tube, oscillating by a vortex oscillator, getting soil suspension into 96 micropore plates (occupies n micropores) under the oscillation condition, adding 4-MUB-7-Beta-D-cellobioside substrate solution into n-1 holes, and adding water with equal quantity into the other one hole so as to be used as non-substrate contrast, adding substrate solution with equal quantity and water with equal quantity into the (n+1)th hole so as to be used as soli-free contrast, oscillating and culturing under constant temperature; secondly, adding NaOH into the micropore plates to terminate the reaction after the culture is finished; thirdly, performing the fluorimetric determination to resultant of reaction by a multifunctional microplate reader; and fourthly, calculating the activity of the xylanase. Compared with the traditional method, the invention shortens the culture time, omits the operation procedures of filtration, and the like and simplifies the operation step; meanwhile, the determination data of fluorescent materials in the micropore plate can be obtained within 15s through the multifunctional microplate reader, huge samples are allowed to be simultaneously determined; and moreover, the invention has high accuracy and easy operation, stable and reliable result and good reproduction quality.

Description

technical field [0001] The invention relates to the measurement of cellulase activity in soil, in particular to an analysis method for detecting cellulase activity in soil. Background technique [0002] Carbohydrates entering and accumulating in the soil participate in the carbon cycle under the action of soil sugar enzymes. Cellulose and hemicellulose are the main types of carbohydrates. The hydrolysis of cellulose to its monomer-glucose is in Under the action of different enzymes of the multi-enzyme system of the cellulase complex, it proceeds through several stages. The cellulase complex in soil is composed of four carbohydrases: 1,4-β-glucanase, 1,4-β-glucanase, 1,4-β-glucoside exonuclease and cellobiase. They are involved in different stages of cellulose decomposition. The soil cellulase activity usually measured is the sum of the effects of the enzyme complex. Cellulase activity is often used as an indicator of soil fertility because it participates in the decompos...

Claims

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Application Information

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IPC IPC(8): C12Q1/34G01N21/64
Inventor 张丽莉武志杰陈利军
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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