• 한국어병학회지
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• 제19권1호
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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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• 제5권3호
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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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• 제14권1호
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• pp.103-108
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• 1999

혈전 용해효소를 생산하는 미생물을 탐색하기 위하여 장류 및 토양사료로부터 미생물을 분리하여 그 중 fibrin 용해활성이 높고 기질특이성이 우수한 S7-l6 균주를 선발하였다. 선발된 미생물은 Gram 양성의 간균으로 운동성이 있고, catalase 양성이었으며 포자를 형성하는 것으로 관찰되어 BClcilius sp. S7-16으로 명명하였다. $\alpha$cill$\mu$ sp S7-16의 효소생산 조건을 검토한 결과,0.5% (w/v) polypeptone, 0.5% (w/v) yeast extract, 0.3% (w/v) NaCI, 0.l%(w/v) $KH_2P0_4$, 0.3% (w/v) $KH_2P0_4$, 0.01% (w/v) $MgSO_4.7H_20$(pH7.0)의 배지조성하에서 $35^{\circ}C$애서 180 rpm으로 24시간 회전진탕배양하였을 때 fibrin 용해효소 생산이 최대를 보였으며, 이때 casein 용해활성은 나타나지 않았다. Fibrin 용해효소의 효소적 특성을 조사하기 위해 이 조건하에서 얻어진 배양상동액에 20~60% ammomum sulfate 분획침전을 행하여 조효소액을 조제하였으며, 얻어진 조효소액은 pH4~11의 범위에서 안정하였으며, $50^{\circ}C$까지 비교적 활성이 유지되는 열안 정성을 보여주었다. 조효소액의 최적 pH와 온도는 각각 7.5, $40^{\circ}C$부근에서 최적활성을 보였다.