Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Real Time Measurement of Protease Activity of Live Uronema marinum (Ciliata: Scuticociliatida) by Fluorescence Polarization Assay

  • Lee Eun Hye (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kwon Se Ryun (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kim Chun Soo (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Chung Joon Ki (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Lee Hyung Ho (Faculty of Food Science and Biotechnology, Pukyong National University) ;
  • Kim Ki Hong (Department of Aquatic Life Medicine, Pukyong National University)
  • Published : 2002.12.01
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Abstract

Proteolytic activity of live Uronema manum was analyzed by fluorescence polarization (FP) technique. Protease activity was measured by a decrease in FP value using fluorescein isothiocynate (FITC)-casein as a protein substrate. The results demonstrated an inverse linear relationship between fluorescence polarization (FP) values and live ciliate concentration over the range $1\times10^4\;to\;2\times10^5$ cells/well. However, the FP values of $10-10^3$ live parasites were not different significantly from that of control. Time-dependent decrease in FP value was shown in the wells containing live U. marinum. In the present study, FP assay had the benefit to provide measurements of substrate hydrolysis by live parasites in real-time, and did not require separations, precipitations, or transfers of reaction mixture.

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References

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