Analyzing method for detecting activity of soil xylanase

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 under the oscillation condition, adding 4-MUB-7-Beta-D-xyloside 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 steps; 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 determination of xylanase activity in soil, in particular to an analysis method for detecting xylanase activity in soil. Background technique [0002] Carbohydrates entering and accumulating in the soil are involved in the carbon cycle under the action of soil sugar enzymes. Xylanase is directly involved in the decomposition and transformation of major polymers in organic components such as plant litter and leaves. It can be used as organic matter to reduce An effective indicator of xylanase activity, usually the measurement of xylanase activity is the sum of intracellular xylanase and extracellular xylanase activities, which decompose xylan into stable xylose that can be absorbed and utilized by plants or other carbohydrates. Xylanase activity is often used as an indicator of soil fertility, because it participates in the decomposition of soil organic matter. In addition, there is a correlation between the activity of xyl...

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Problems solved by technology

[0003] The determination of relevant soil xylanase activity basically all refers to the method (Li Fudi, Yu Ziniu, He Shaojiang. Agricultural Microbiology Experimental Technology. Beijing : China Agricultural Press, 1996.36: 305-308) and the method described in the monograph Methods in Soil Biology written by Schinner and van Mersi in 1996, these two methods all use spectrophotometry to measure the production amount of the final product to characterize Enzyme activity, the detection sensitivity of spectrophotometry is relatively low, so the requirements for the concentration of the analyte are relatively high. Both of these two methods have a long incubation time for the reaction system and require more complicated determination steps such as filtration (centrifugation), Li Fu The chromogenic substances obtained by the method used in Di’s paper have low stability and poor measurement reproducibility. Although the method described by Schinner and van Mersi overcomes this defect, the measurement process is also complicated and takes a long time. , and the reagent potassium cyanide required in the color development process is a highly toxic reagent, which is difficult to obtain and increases the danger to the operator during the determination process

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