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. 2018 May 20;8(10):e2856.
doi: 10.21769/BioProtoc.2856.

Enzymatic Activity Assay for Invertase in Synechocystis Cells

Xiaoming Tan  1 , Kuo Song  1 , Xuefeng Lu  1   2
Affiliations

Enzymatic Activity Assay for Invertase in Synechocystis Cells

Xiaoming Tan et al. Bio Protoc. .

Abstract

Invertase can catalyze the hydrolysis of sucrose, and is widely distributed in cells of cyanobacteria and plants. Being responsible for the first step for sucrose metabolism, invertase plays important physiological roles and its enzymatic activity is frequently needed to be determined. All the methods for determination of the invertase activity are dependent on detection of the glucose product generated by the invertase. Here we describe an ion chromatography based protocol of our laboratory for determination of cyanobacterial intracellular invertase activity.

Keywords: Cyanobacteria; Invertase enzymatic activity; Ion chromatography; Sucrose; Synechocystis.

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Figures

Figure 1.
Figure 1.. Ion chromatography profiles of standards and the in vitro invertase reaction mixtures.
A. Chromatogram of the glucose-fructose standard mixture. 100 mg/L of the glucose and fructose standards were mixed together and analyzed by ion chromatography. B. Chromatogram of the in vitro invertase reaction mixture without the cell crude extract (negative control). C. Chromatogram of the in vitro invertase reaction mixture. Glu, Glucose; Fru, Fructose; Suc, Sucrose.
Figure 2.
Figure 2.. The glucose standard curve
See this image and copyright information in PMC

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