Direct colorimetric assay of microcystin using protein phosphatase |
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Authors: | Hee-Mock Oh Seog June Lee Jee-Hwan Kim Chan-Sun Park Byung-Dae Yoon |
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Institution: | (1) Environmental Bioresources Laboratory, Korea Research Institute of Bioscience and Biotechnology, Yusong, P.O. Box 115, 305-600 Taejon, Korea |
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Abstract: | A new direct colorimetric assay of microcystin in water and algal samples is proposed consisting of two procedures as follows:
1) the elimination of phosphorus in the sample and concentration of microcystin using a C18 cartridge, 2) the detection of the released phosphorus by the ascorbic acid method and determination of protein phosphatase
(PP) inhibition by microcystin. The optimum amounts of phosphorylase α and PP-1 in 50 μL concentrated sample were 50 μg/50
μL buffer and 1.0 unit/50 μL buffer, respectively, for the best assay. The pH for the maximum activity of PP-1 was 8. The
minimum detectable concentration for this method was about 0.02 μg/L, which is sufficient to meet the proposed guideline level
of 1 μg microcystin/L in drinking water. Consequently, it would seem that the proposed direct colorimetric assay using PP
is a rapid, easy, and convenient method for the detection of microcystin in water and algal samples. |
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Keywords: | ascorbic acid method direct colorimetric assay microcystin protein phosphatase inhibition |
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