Method for detecting and analyzing activity of key enzyme in sugar metabolism of fruits

Abstract

The invention provides a method for detecting and analyzing activity of key enzymes in sugar metabolism of fruits. The method is used for determining the activity of fructose synthetase, glucose synthetase, acid invertase and neutral invertase in the fruits. The method comprises the following steps: firstly, preparing a sample, selecting fruits in a specific growth stage, cleaning, grinding and centrifuging to obtain a crude enzyme extracting solution; and then, designing a specific reaction system aiming at different enzymes, simulating enzymatic reaction conditions, utilizing a substrate and an enzyme reaction product, and quantitatively determining the product by adopting a high-sensitivity method. And finally, carrying out statistical analysis on enzyme activity data, calculating an average value and a standard deviation, and carrying out comparison. According to the method, reaction system parameters are optimized, the fruit physiological environment is simulated, quality control is strict, and the detection accuracy is remarkably improved; the method is simple and convenient to operate, low in cost and wide in application range, can simultaneously detect activity of various enzymes, and provides powerful support for comprehensive understanding of a fruit sugar metabolism mechanism, regulation and control of fruit quality and variety improvement.

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