Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Proteases in Cell Lysate of Uronema marinum (Ciliata: Scuticociliatida), an Opportunistic Pathogen of Cultured Olive Flounder (Paralichthys olivaceus)

  • Kwon Se Ryun (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kim Chun Soo (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Ahn Kyoung Jin (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Cho Jae Bum (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 : 2002.09.01
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Abstract

The effects of pH, temperature and various inhibitors on the proteolytic activity of the cell lysate of Uronema marium were investigated using colorimetric and substrate gel electro­phoretic methods. The cell lysate of U. marinum showed proteolytic activity over a wide range of pH, and pH optima ranged from pH 5 to 7. The proteolytic activity was increased according to a rise of temperature but decreased at $40^{\circ}$. The proteolytic activity of the parasite lysate was significantly inhibited by protease inhibitors including trans-epoxysuccinyl -L-leucylamido-(4-guanidino) butane (E-64), pepstatin A, phenyl-methanesulfonyl fluoride(PMSF), and ethylenediamine-tetraacetic acid (EDTA). Preincubation of the lysate with E-64 showed the maximum inhibition of the caseionolytic activity. Four protease bands (152, 97, 67 and 40 kDa) were detected by gelatin SDS-PAGE. Significant inhibition of caseinolytic activity and complete abolition of a 152 kDa band in gelatin SDS-PAGE by EDTA indicated that the cell lysate of U. marinum had a metalloprotease Another three proteolytic bands were inhibited by E64, a cysteine protease inhibitor. Preincubation of the cell lysate with pepstatin or PMSF had no effects on the protease bands.

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References

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