Lipolytic 효소를 생산하는 Acinetobacter sp. BD5 균주의 분리 및 특성 = Isolation and Characterization of Acinetobacter sp. BD5 Producing Lipolytic Enzyme

국문 초록 (Abstract)

유용효소를 생산하는 균주를 가정하수로부터 분리하기 위하여 LBM, R2A, M9배지를 이용하여 다수의 균주를 분리하였다. 분리된 균주 중 1% tributyrin이 첨가된 배지에서 생육 활성대의 형성이 우수한 균주 1종을 최종적으로 선별하여 형태학적, 생리학적, 생화학적 특성을 관찰하였다. 16S rDNA 염기서열 분석결과 Acinetobacter Baumannii (99%)로 Acinetobacter 속에 속하는 균주임을 확인하고, Acinetobacter sp. BD5로 명명하였다. Acinetobacter sp. BD5는 $37^{\circ}C$와 $50^{\circ}C$에서 생육하는 것으로 보아 호열성 균주이며, 1% tributyrin과 oilve oil 첨가된 EL과 CE 고체배지와 Tween 20이 첨가된 LB 고체배지에서 생육활성대의 형성을 확인하여 이 균주가 lipolytic 효소를 생산하는 것으로 나타났다. 효소 활성의 최적 배양조건을 검토하고자 배양시간별 효소 활성을 측정한 결과, 배양 6시간 때인 대수 증식기에 가장 높은 효소활성을 나타내어 비교적 빠른 시간 내에 lipolytic 효소를 생성하는 것으로 나타났다. 또한 효소활성의 최적 온도는 $60^{\circ}C$로 $70-80^{\circ}C$에서 70%이상의 잔존활성을 보이는 것으로 나타나 Acinetobacter sp. BD5는 호열성 균주로 이 균주가 생산하는 lipolytic 효소도 내열성을 보이는 것으로 생각된다. 최적 pH는 9.0이며, pH 9.8-10.6범위에서 50% 이상의 활성이 유지되어 alkaline lipolytic 효소인 것으로 생각되어진다. Acinetobacter sp. BD5가 생산하는 lipolytic 효소는 비교적 넓은 pH 범위와 고온에서 활성이 유지되는 것으로 보아 폐유분해, 세제 합성과 유기물질 합성 등 생물공학분야와 산업적으로 잠재적 가치가 있을 것으로 생각된다.

다국어 초록 (Multilingual Abstract)

A bacterium producing novel lipolytic enzyme was isolated from house sewage and identified as Acinetobacter sp. BD5 based on physiological characterization and 16S rDNA sequencing. The lipolytic activity of Acinetobacter sp. BD5 was tested using an EL agar medium and CE agar medium supplemented with 1% tributyrin and olive oil, respectively. The formation of a clear zone around the colony was detected by agar medium supplemented with 1% tributyrin and olive oil, respectively and Acinetobacter sp. BD5 formed powder-like zone around the colony on LB agar medium containing Tween 20. The quantitative lipolytic activity was determined by using p-NP butyrate as substrate. Acinetobacter sp. BD5 secreted the lipolytic enzyme during exponential growth phase, reaching a maximum amount after 6 hours of incubation. The lipolytic enzyme was found to be optimally active at $60^{\circ}C$ and retained more than 70% at $70-80^{\circ}C$. It displayed a high degree of activity in a pH of 7.0 to 10.6, with an optimal pH of 9.0.

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