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A quantitative assay for agarase activity determination using agarose-iodine complex
Sug-Young Yoon1, Gi-Hyeon Chae1, Si-Wook Jang1
1Biomolecular Chemistry Laboratory, Department of Chemistry, College of Natural Sciences, Chung-Ang University, 84, Heukseok-Ro, Dongjak-Gu, Seoul, 06974, South Korea.
Abstract:
Rapid and simple spectrophotometric methods are required to detect various oligosaccharides produced by agar-hydrolysing enzymes. Herein, we present a quantitative agarose-iodine assay for agarase activity determination via the detection of the extent of agarose degradation. The agarose-iodine complex becomes reddish orange upon the addition of Lugol solution, and the enzymatic activity can be detected with ultraviolet-visible spectroscopy at 600 nm. The main advantages of this modified Lugol assay are high sensitivity, simple detection, and cost effectiveness. A novel definition of the unit to measure and compare the activities of agarases is also suggested.
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