| Project/Area Number |
11558076
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
環境保全
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| Research Institution | Toyohashi University of Technology |
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Principal Investigator |
NISHIMURA Kazuyuki Toyohashi University of Technology, Dep.of eng., Associate Professor, 工学部, 助教授 (00261595)
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| Co-Investigator(Kenkyū-buntansha) |
宮本 信一 新日本気象海洋株式会社, 環境創造研究所・バイオ技術グループ, 職員(研究職)
KUNIKANE Shoichi National Institute of Public Health, Dep.of Water supply eng., chief director, 水道工学部, 部長(研究職) (90083740)
MIYAMOTO Nobukazu METOCEAN Consultant Co.Ltd, Inst.of Environ.Ecology, Researcher
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 2000: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Microcystin / protein phosphatase / Immobilization of protein phosphatase / Monitoring system / Rapid Analysis / Monitoring / モニタリング装置 |
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| Research Abstract |
The fast analysis of microcystin in environmental water was examined by using a protein phosphatase 2A inhibition assay. The heat treatment method was developed for rapid extraction of microcystin from Microcystis cells.
Quantitative analysis of microcystin by the protein phosphatase 2A inhibition assay required two hours. The assay was accurate and reproducible. The cultured Microcystis suspension was submitted to the protein phosphatase 2A inhibition assay as well as commercial ELISA assay kit with similar results. But sensitivity of this assay was not so good as the ELISA assay kit. The microcystins were extracted from Microcystis cells by heat conditions such as temperature of 50-60℃ and heating time of more than 20 minutes. The heat treatment with-amylase, cellulase and lysozyme was slightly facilitated to extraction of it.
Uses of the protein phosphatase 2A inhibition assay with the heat treatment method facilitate investigation of microcystin in natural water.
The Monitoring system for Microcystin was made in three parts, such as extraction, concentration and detection equipment.
The microcystins were extracted from Microcystis cells by heat treatment in extraction part. Extracted microcystins were concentrated by ultra-filtration in concentration part. And, those were determined by inhibition assay use of immobilization of protein phosphatase in detection part.
In this study, any type microcystins was not detected in samples of resource water for drinking and samples of tap water. And, high concentration of microcystins was detected in samples of pond for agriculture. Concentration of microcystins was not related with Microcystis cell Numbers and/or water temperature.
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