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KMID : 1094720000050060418
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Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 6 p.418 ~ p.421
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Direct colorimetric assay of microcystin using protein phosphatase
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Oh Hee-Mock
Lee Seog-June
Kim Jee-Hwan
Park Chan-Sun
Yoon Byung-Dae
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Abstract
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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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KEYWORD
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ascorbic acid method, direct colorimetric assay, microcystin, protein phosphatase inhibition
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