• Journal of fish pathology
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• v.19 no.1
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• pp.83-86
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• 2006

Effect of iron on the extracellular proteolytic activity of live Uronema marium was determined by fluorescence polarization (FP) method. Supplementation of 0.5 and 5.0 μM iron significantly increased caseinolytic activity of live U. marinum. In contrast, supplementation of 50 μM iron showed no significant differences in FP values compared to the control. The present result suggests that iron in cultured water or skin tissue of olive flounder may influence on the penetration and establishment of U. marinum, correlating with modulation of extracellular protease activity of the ciliates.

• Fisheries and Aquatic Sciences
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• v.5 no.3
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• pp.145-149
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• 2002

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.

• KSBB Journal
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• v.14 no.1
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• pp.103-108
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• 1999

A bacterium having strong fibrinolytic activity, S7-16 strain, was isolated from soil. The isolated bacterium was identified and named as Bacillus sp. S7-16. The optimal composition of the medium for the production of fibrinolytic enzyme by Bacillus sp. S7-16 was 0.5%(w/v) polypeptone, 0.5%(w/v) yeast extract, 0.3%(w/v) NaCl, 0.1% (w/v) $KH_2PO_4,\;0.3%(w/v)\;K_2PHO_4,\;and\;0.01%(w/v)\;MgSO_4{\cdot}7H_2O$. The optimal temperature and initial pH of the medium for the production of the enzyme were $35^{\circ}C$ and 7.0, respectively. The maximum production of the fibrinolytic enzyme was obtained after 24 hours of the incubation. Under the above conditions, the culture supernatant had strong fibrinolytic activity. Within pH4~11, the crude fibrinolytic enzyme was stable. The enzyme was stable up to $50^{\circ}C$. The optimum pH and temperature for the enzyme activity were around 7.5 and $40^{\circ}C$, respectively.