Kit and method for detecting activity of sucrose phosphate synthase

Abstract

The invention discloses a kit and method for detecting the activity of sucrose phosphate synthase. The kit comprises an extracting solution prepared from Tris-HCl, magnesium chloride, EDTA and 2-mercaptoethanol, a reagent I prepared from Tris-HCl, magnesium chloride, 6-phosphofructose and UDPG, a reagent II prepared by dissolving saccharose in distilled water, a reagent III prepared by dissolving sodium hydroxide in distilled water, a reagent IV prepared by dissolving hydrochloric acid in distilled water and a reagent V prepared by dissolving resorcinol in ethanol and distilled water. According to the method, the influence of saccharose in a sample can be eliminated, so that the detection result is accurate, and the kit is suitable for detecting the activity of sucrose phosphate synthase in various samples. According to the kit and method, the number of reagent types can be greatly reduced, the operation can be simple and convenient, the total volume of the reaction is only 300mu L, the concentration of UDPG can be reduced on the base of not influencing the accuracy, and the reagent cost can be greatly reduced.

Description

technical field [0001] The invention belongs to the field of life sciences and relates to a kit, in particular to a kit for measuring the activity of sucrose phosphate synthase and a method thereof. Background technique [0002] Sucrose is not only an important photosynthetic product, but also the main substance transported in plants and one of the storage forms of carbohydrates. Sucrose phosphate synthase (SPS) accepts fructose-6-phosphate, and the formed sucrose phosphate is converted into sucrose under the action of sucrose phosphatase. Generally, the sucrose phosphate synthase-sucrose phosphatase system is regarded as the main pathway of sucrose synthesis. [0003] The currently available method for detecting the activity of sucrose phosphate synthase is the resorcinol colorimetric method. The principle is that sucrose phosphate synthase catalyzes fructose-6-phosphate to form sucrose phosphate in the sample, and the depth of the color change presented by the reaction b...

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Problems solved by technology

[0004] 1. It is not suitable for the detection of samples with high sucrose content. The sucrose contained in the sample will have a great impact on the determination of the enzyme activity, resulting in inaccurate determination results

[0005] 2. The total reaction volume is 2-3mL, and the amount of various reagents added is relatively large, especially the concentration of UDPG is above 10mM. The reagent is expensive and the overall cost is high

[0006] 3. There are too many types of reagents and the operation process is too cumbersome

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