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Assessing the microcystins concentration through optimized protein phosphatase inhibition assay in environmental samples
Kyoung-Hee Oh , Kung-Min Beak , Yuna Shin , Young-Cheol Cho
J. Microbiol. 2022;60(6):602-609.   Published online April 30, 2022
DOI: https://doi.org/10.1007/s12275-022-2020-4
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AbstractAbstract
Protein phosphatase (PPase) inhibition assay (PPIA) is widely used to analyze the concentration of microcystins (MCs) because it is comparatively less expensive and faster than other assays. This study aimed to optimize the PPIA by determining a suitable reaction terminator and an optimal methanol concentration in the sample. The most suitable reaction time was 90 min, with the corresponding methanol concentration in the sample being 15% or less. When p-nitrophenyl phosphate (pNPP) was used as a substrate, copper chloride solution was suitably used as a reaction terminator, and when 4- methylumbelliferyl phosphate (MUP) was used, a glycine buffer not only increased the measurement sensitivity of the reaction product but also terminated the enzymatic reaction. When PPase 1 and MUP were used as an enzyme and a substrate, respectively, the limit of quantitation for MC-leucine/ arginine (LR) was 0.02 μg/L, whereas it was 0.1 μg/L when pNPP was used as a substrate. The proposed method facilitated the measurement of MC-LR concentration without additional pretreatments, such as concentration or purification; therefore, this method was suitable and feasible for the continuous monitoring of MCs in drinking water.

Citations

Citations to this article as recorded by  
  • Analyzing MC-LR distribution characteristics in natural lakes by a novel fluorescence technology
    Xiangyu Hu, Zhaomin Wang, Xiao Ye, Ping Xie, Yong Liu
    Environmental Pollution.2024; 342: 123123.     CrossRef
  • Magnetic solid phase extraction coupled with high-performance liquid chromatography-diode array detection based on assembled magnetic covalent organic frameworks for selective extraction and detection of microcystins in aquatic foods
    Tianliang Wang, Hongzhen Xie, Yuting Cao, Qing Xu, Ning Gan
    Journal of Chromatography A.2022; 1685: 463614.     CrossRef

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