Bacillus sp. DYL130 균주의 Biosurfactant의 정제 및 특성

Purification and Characterization of Biosurfactant from Bacillus sp. DYL130

  • 박인혜 (동아대학교 응용생명공학부) ;
  • 김선희 (동아대학교 응용생명공학부) ;
  • 이상철 (동아대학교 응용생명공학부) ;
  • 하순옥 (동아대학교 응용생명공학부) ;
  • 이용석 (동아대학교 응용생명공학부) ;
  • 유아름 (동아대학교 응용생명공학부) ;
  • 김근기 (부산대학교 생명응용과학부) ;
  • 최용락 (동아대학교 응용생명공학부)
  • Park, In-Hye (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Kim, Sun-Hee (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Sang-Cheol (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Ha, Soon-Ok (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Lee, Yong-Seok (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Ryu, Ah-Reum (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University) ;
  • Kim, Keun-Ki (Department of Life Science and Environmental Chemistry, Pusan National University) ;
  • Choi, Yong-Lark (Department of Biotechnology, College of Natural Resources and Life Science, Dong-A University)
  • 발행 : 2006.12.31

초록

해양 원유 오염원의 제거 및 항진균 활성을 가진 균주를 분리하기 위해 1% crude oil을 탄소원으로 이용하여 생육하는 수십 종의 균주를 분리하였다. 분리된 균주 중 biosurfactant를 생성하는 균주를 1% tributyrin에서 투명환 생성 유무를 통해 선별되었으며, 이를 Bacillus sp. DYL130으로 명명하였다. Bacillus sp. DYL130의 표면장력은 최저 28 mN/m까지 감소되었다. 이 균주로부터 biosurfactant를 정제하기 위해 배양 상등액을 친화성 크로마토그래피와 TLC를 이용하여 정제된 biosurfactant를 얻었다. TLC 결과 0.78정도의 Rf값을 확인하였다. 정제된 biosurfactant의 CMC 값은 35 mg/l인 것으로 나타났으며, 유화활성과 안정성은 soybean oil을 기질로 사용하였을 경우 최대인 것으로 나타났다. Botrytis cineria에 대한 길항작용도 우수한 것으로 보인다.

Bacillus sp. DYL130 producing biosurfactant was isolated from soil samples in the Duck-yu mountain and identified as Bacillus sp. by analysis of 16S rDNA sequence. Purification of the biosurfactant was performed by using affinity chromatography and TLC. The biosurfactant of culture medium from Bacillus sp. DYL130 was eluted with 100% methanol using affinity chromatography. To remove methanol, a rotary evaporator was used and enrichment sample was dissolved in alkaline water(pH 10). The purified biosurfactant was identified by TLC. It was confirmed that the Rf value of the biosurfactant was 0.78. Antifungal activity against Botrytis cineria was showed the strongly activity as active antagonist. Maximum emulsification activity and stability were obtained from soybean oil. The critical micelle concentration (CMC) of purified biosurfactant was 35mg/l and the purified biosurfactant inhibited biofilm forming by Bacillus sp..

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