Improved determination method of nitrite reductase

Abstract

The invention discloses an improved determination method of nitrite reductase in soil. In the presence of a hydrogen donor and under anaerobic conditions, nitrite reacts with a sulfanilic acid-alpha-naphthylamine reagent to determine the variation of NO2<->-N before and after enzymatic reaction so as to represent the activity of nitrite reductase. A vacuum dryer is adopted to replace a decompression triangular flask to serve as a culture container, so that the culture environment is anaerobic, the operation process can be simplified, and data errors caused by culture condition differences arereduced. Meanwhile, a certain amount of KClO3 is added during testing and standard curve preparation, so that NO2<-> can be inhibited from being oxidized into NO3<->, the loss of NO2<-> is reduced, and the data accuracy is improved. The method disclosed by the invention is high in reproducibility, simple and feasible, and capable of reducing errors, so that the stability of a measurement result between parallel processing is higher, and the measurement result is closer to a real value.

Description

technical field [0001] The invention relates to the technical field of measuring soil enzyme activity, in particular to an improved measuring method for nitrite reductase. Background technique [0002] Under the action of nitrate reductase, nitrite reductase and hydroxylamine reductase, nitrate nitrogen in soil can be reduced to ammonia. Determination of the activity of these enzymes in soil can understand the intensity of deamination in soil nitrogen transformation. In addition, studying the activity of nitrite reductase in soil is of great significance for rational fertilization and reducing the loss of soil nitrogen. [0003] In the traditional nitrite reductase assay method, in order to ensure that the soil culture condition is an anaerobic environment, the soil is cultivated in a decompression triangular flask, and the decompression triangular flask is pumped during cultivation. In the actual operation process, when the number of samples is large, the pumping process ...

AI Technical Summary

Problems solved by technology

In the actual operation process, when the number of samples is large, the pumping process takes a long time, which not only delays the progress of the experiment, but also causes errors in the experimental results due to asynchronous sample processing.

And it is difficult to guarantee the tightness of the decompression triangular flask, which is also one of the reasons for the experimental error

At the same time, the measured enzyme is NO in soil 2 - Convert to NH 2 Catalytic enzyme for OH, NO 2 - The amount of loss represents the activity of nitrite reductase, while NO 2 - easily oxidized to NO 3 - (NO 2 - +0.5O 2 →NO 3 - ), resulting in a large discrepancy between the measured results

Therefore, the repeatability of the final test results is poor, and it is necessary to improve the assay method of nitrite reductase, reduce the experimental error, reduce the deviation between the measured results, and increase the repeatability of the experiment.

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