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An analytical method for detecting soil cellulase activity

An analysis method, cellulase technology, applied in the direction of material stimulation analysis, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem 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: 2012-02-08
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • An analytical method for detecting soil cellulase activity
  • An analytical method for detecting soil cellulase activity

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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 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 analysis method for detecting cellulase activity in soil: 1) Take n parts of sieved soil air-dried samples in n thick test tubes, add acetic acid buffer solution to each test tube, vibrate with a vortex oscillator, Take the soil suspension to a 96-microwell plate (occupying n microwells) under shaking conditions, add 4-MUB-7-β-D-cellobioside substrate solution to n-1 wells, and add 4-MUB-7-β-D-cellobioside substrate solution to another well Add an equal amount of water as a no-substrate control, add an equal amount of substrate solution and an equal amount of water into the n+1th well as a no-soil control, and culture with shaking under constant temperature conditions; 2) Add NaOH terminates the reaction; 3) Fluorescent determination of the reaction product by a multifunctional microplate reader; 4) Calculation of cellulase activity. The advantages of the present invention are: 1) Compared with the traditional method, the culture time is shortened, operating procedures such as filtration are omitted, and the operation steps are simplified; The measurement data can be obtained at the same time in the same time, allowing a large number of samples to be measured at the same time; 3) High accuracy and easy operation; 4) The results are stable and reliable, and the reproducibility is good.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34G01N21/64
Inventor 张丽莉武志杰陈利军
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI