Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Culture Conditions for Improving Extracellular Lipolytic Enzyme Production by a Novel Thermophilic Geobacillus sp. AR1

신규 고온성 Geobacillus sp. AR1의 extracellular 지질분해효소 생산을 위한 배양조건

  • Park, Su-Jin (Department of Biotechnology & Bioengineering, Dong-Eui University) ;
  • Jeon, Sung-Jong (Department of Biotechnology & Bioengineering, Dong-Eui University)
  • Received : 2012.11.14
  • Accepted : 2013.01.04
  • Published : 2013.01.30
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Abstract

A microorganism (strain AR1) producing an extracellular lipolytic enzyme was isolated from hot springs located in Beppu, Japan. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that AR1 belongs to the genus Geobacillus. This study focused on novel strategies to increase extracellular lipolytic enzyme production by this novel Geobacillus sp. AR1. Cultures of the AR1 strain grew within a wide temperature range (from 35 to $75^{\circ}C$); the optimum temperature was $65^{\circ}C$. The pH for optimal growth was 6.5, whereas the optimum pH for lipolytic enzyme production was 8.5. The presence of oils in the culture medium led to improvements in lipolytic enzyme activity. Soybean oil was the most efficient inducer, and it yielded better results when added in the exponential phase. On the other hand, the addition of chemical surfactants led to lipolytic enzyme production. Their addition to the culture could affect the location of the enzyme activity. The addition of Tween 20 in the stationary phase significantly increased the proportion of the extracellular enzyme activity. According to the results, following the addition of soybean oil and Tween 20 in the exponential and stationary phases, the extracellular lipolytic activity was increased 2.4-fold compared with that of a control.

Extracellular 지질분해효소를 생산하는 균주 AR1은 일본 벳부 온천수에서 분리하였다. 분리된 균주의 16s rRNA 염기서열을 분석하고 계통학적으로 분류한 결과, AR1 균주는 신규 Geobacillus sp.에 속하는 것으로 동정되었다. 본 연구는 Geobacillus sp. AR1 균주의 extracellular 지질분해효소 생산을 향상시키기 위한 새로운 방법에 초점을 맞추었다. AR1 균주는 $35{\sim}75^{\circ}C$의 넓은 온도 범위에서 생육하였고 최적온도는 $65^{\circ}C$이었다. 생육을 위한 최적 pH는 6.5인 반면, 효소 생산을 위한 pH는 8.5로 차이점을 보였다. 배양 중에 지질 화합물의 첨가는 지질분해효소 생산을 유도하였고, soybean oil을 대수증식기에 첨가 했을 때 가장 효율적인 유도 효과를 나타내었다. 한편, 계면활성제는 지질분해효소의 생산을 유도하고 세포 내외의 위치에 영향을 줄 수 있다. AR1 균주는 정지기에 Tween 20을 첨가할 경우, 효소의 세포 외 분비 효율이 크게 증가하였다. 이들 결과를 바탕으로 soybean oil과 Tween 20을 각각 대수증식기와 정지기에 첨가함에 따라 extracellular 효소 생산이 대조구에 비해 2.4배 증가하는 것으로 확인 되었다.

Keywords

References

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