DOI QR코드

DOI QR Code

Bacillus atrophaeus DYL-130이 생산하는 biosurfactant의 분리 및 특성

Isolation and Characterization of Biosurfactant from Bacillus atrophaeus DYL,-130

  • 김선희 (동아대학교 응용생명공학부) ;
  • 이상철 (동아대학교 용용생명공학부) ;
  • 박인혜 (동아대학교 응용생명공학부) ;
  • 유주순 (동아대학교 응용생명공학부) ;
  • 주우홍 (창원대학교 생물학과) ;
  • 황철원 (한동대학교 생명공학 연구소) ;
  • 최용락 (동아대학교 응용생명공학부)
  • Kim Sun-Hee (Department of Biotechnology, Faculty of Natural Resources and Life Science Dong-A University) ;
  • Lee Sang-Cheol (Department of Biotechnology, Faculty of Natural Resources and Life Science Dong-A University) ;
  • Park In-Hye (Department of Biotechnology, Faculty of Natural Resources and Life Science Dong-A University) ;
  • Yoo Ju-Soon (Department of Biotechnology, Faculty of Natural Resources and Life Science Dong-A University) ;
  • Joo Woo-Hong (Dept. of biology, Chang Won University) ;
  • Hwang Cher-Won (Institute of Bioscience and Technology, Handong Global University) ;
  • Choi Young-Lark (Department of Biotechnology, Faculty of Natural Resources and Life Science Dong-A University)
  • 발행 : 2005.10.01

초록

원유 분해능이 강력한 균주를 얻고자 덕유산의 토양으로부터 crude oil을 탄소원으로 이용하는 수십 종을 분리하였다. 분리된 균주 중 원유분해능 및 biosurfactant 생성능이 우수한 균주를 선별하여, 형태학적 특성을 관찰하고 165 rDNA sequence와 』gyrA gene sequence를 통하여 Bacillus atrophaeus DYL-130으로 동정하였다. 동정된 균주 배양액의 표면장력은 최저 28 mN/m까지 감소되었다. 또한 Bacillus atrophaeus DYL-130이 생산하는 biosur-factant가 column chromatography에 의하여 분리 되었으며, 분리된 biosurfactant의 toluene 유화능 및 crude biosurfactant의 유화활성과 안정성이 시험 되었다. Crude biosurfactant의 유화활성은 kerosene에서 최대였으며, soybean oil에서도 높은 편이였으나 유화안정성의 경우 soybean oil을 기질로 하였을 경우가 kerosene을 기질로 하였을 때 보다 우수하였다. 또한 crude biosurfactant의 유화안정성을 합성계면활성제와 비교 한 결과 DYL-130이 생산하는 biosurfactant의 유화안정성이 합성계면활성제와 유사하거나 뛰어남을 확인하였다. Column chroma-tography에 의하여 분리된 biosurfactant는 drop-collapsing method에 의하여 surface-activity가 확인 되었으며 또한 toluene에 대한 유화력이 매우 뛰어난 것을 확인 하였다. 따라서 DYL-130에서 추출한 biosurfactant는 합성계면활성제를 대체할 수 있는 환경친화적인 생물 계면활성제로 사용될 수 있는 가능성을 보여주고 있다. 산업적으로 이 물질을 이용하기 위해서 Bacillus atrephaeus DYL-130이 생산하는 biosurfactant에 대한 구조분석과 물리 화학적 특성, 생분해도 및 환경독성 등의 조사를 수행하여야 할 것으로 생각된다.

The objective of this study was investigate the characteristic of biosurfactant produced from the iso-lated strain. The strain was isolated from soli samples of Duck-Yu Mountain and it was identified as Bacillus atrophaeus DYL-130 by 16S rDNA and gyrA gene nucleotide sequence analysis. The surface ten-sion of culture filtrate of Bacillus atrophaeus DYL-130 decreased to 28 mN/m and its biosurfactant con-centration was determined by diluting the culture filtrate until the critical micelle concentration (CMC). The emulsifying activity and stability of crude biosurfactant was measured by using water-immiscible hydrocarbons and oils as substrate. The biosurfactant was purified by affinity chromatography and the surface activity of purified biosurfactant was measured by drop-collapsing method and it could be effectively emulsify toluene.

키워드

참고문헌

  1. Arima, K., A. Kakinuma and G. Tamura. 1968. surfactin, a crystalline peptide lipid surfactant produced by Bacillus subtilis : isolation, characterization. Biochem. Biophys. Res. Comm. 31, 488-49 https://doi.org/10.1016/0006-291X(68)90503-2
  2. Banat, I. M., R. S. Makkar, S. S. Cameotra. 2000. Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol., Rev. May. 53(5), 495-508 https://doi.org/10.1007/s002530051648
  3. Cha, J. Y., B. G. Kim, S. Y. Chung, Y. S. Cho, Y. J. Choi and Y. C. Lee. 1999. Characterization of crude oil degradation by Klebsiella sp. KCL-1 isolated from sea water. Kor. J. Appl. Microbiol. Biotechnol. 27, 452-457
  4. Cirigliano, M. C. and G. M. Carman. 1984. Isolation of a bioemulsifier from Candida lipolytica. Appl. Environ. Microbiol. 48, 747-750
  5. Cirigliano, M. C. and G. M. Carman. 1985. Purification and Characterization of Uposan, a bioemulsifier from Candida lipolytica. Appl. Environ. Microbiol. 50, 846-850
  6. Cooper, D. G. and J. E. Zajic. 1980. Surface-active compounds from microorganism. Adv. Appl. Microbiol. 26, 229-256 https://doi.org/10.1016/S0065-2164(08)70335-6
  7. Desai, J. D. and I. M. Banat. 1997. Microbial production of surfactants and their commercial potential. Microbiol. Mol. BioI. Rev. 61, 47-64
  8. Fiechter, A. 1992. Biosurfactant : moving towards industrial application. Trends Biotechnol. 10, 208-217 https://doi.org/10.1016/0167-7799(92)90215-H
  9. Fiedler, H. P. and W. Umbach. 1987. Cosmetics and toiletries. In 'surfactants in Consumer Products: Theory, Technology and Application', pp.350-398, eds., Falbe, J., Springer Verlag, Heidelberg
  10. Greek, B. F. 1991. Sales of detergents growing despite recession. Chem. Eng. News. 69, 25-52
  11. Greek, B. F. 1990. Detergent industry ponders products for new decade. Chem. Eng. News. 68, 37-38
  12. Kuiper J., E. J. Lagendijk, R. Pickford, J. P. Derrick, G. E. Lamers, J. E. Thomas-Oates, B. J. Lugtenberg, G. V. Bloemberg. 2004. Characterization of two Pseudomonas putida lipopeptide biosurfactants, putisolvin I and II, which inhibit biofilm formation and break down existing biofilms. Mol Microbiol. 51, 97-113 https://doi.org/10.1046/j.1365-2958.2003.03751.x
  13. Kim, H. J., B. J. Kim, S. H. Hwang, D. J. Kim, H. W. Lee and J. Y. Kong. 1997. Degradation of crude oil and purification of biosurfactant from marine bacterium Aeromonas sp. BES-741. Kor. J. Biotechnol. Bioeng. 24, 443-448
  14. Kim, S. H., E. J. Urn, S. O. Lee, J. D. Lee and T. H. Lee. 2000. Purification and characterization of biosurfactants from Nocardia sp. L-417. Biotech. Appl. Biochem. 31, 249-253 https://doi.org/10.1042/BA19990111
  15. Kim, S. H., B. D. Yoon, C. H. Lee, H. Y. Seo, H. M. Oh, T. Katsuragi and Y. Tani. 1997. Production and properties of a lipopeptide biosurfactant from Bacillus sutilis C9. J. Ferment. and Bioengin. 18, 71-75
  16. Kosaric, N. 1993. Biosurfactants : production, property, application. In 'Surfactant Science Series' vol. 48, Marcel Dekker, New York
  17. Lee, S. c., Y. J. Jung, J. S. Yoo, Y. S. Cho, J. H. Cha and Y. J. Choi. 2002. Characteristics of biosurfactants produced by Bacillus sp. LSC11. Kor. J. Life Science. 12, 6, 745-751 https://doi.org/10.5352/JLS.2002.12.6.745
  18. MacFaddin, J. F. 1984. Biochemical Tests for Identification for Medical Bacteria, 2nd ed. Williams and Wilkins Co., Baltimore, USA
  19. Muller-Hurtig, R, R Blaszczyk, F. Wagner and N. Kosaric. 1993. Biosurfactants for environmental control. In 'Biosurfactants for environmental control. In 'Biosurfactants : Production, Properties Applications', pp.447-470, eds., Kosaric, N., Marcel Decker, New York
  20. Schulz, D., A. passeri, M. Schmidt, S. Lang, F. Wanger, V. Wray and W. gunkel. 1991. Marine biosurfactants, J. Screening for biosurfactants among crude oil dergading marine microorganism from the North Sea. Z. Naturforsch. 46, 167-203
  21. Zhajic, J. E., H. Gingnard, D. F. Gerson. 1977. Properties and biodegradation of a bioemulsifier from Corynebacterium hydrocarboclastus. Biotechnol. Bioeng. 19, 1303-1320 https://doi.org/10.1002/bit.260190905

피인용 문헌

  1. A halotolerant, thermotolerant, and facultative biosurfactant producer: Identification and molecular characterization of a bacterium and evolution of emulsifier stability of a lipopeptide biosurfactant vol.16, pp.1, 2011, https://doi.org/10.1007/s12257-010-0148-2
  2. Isolation, purification, and characterization of novel fengycin S from Bacillus amyloliquefaciens LSC04 degrading-crude oil vol.15, pp.2, 2010, https://doi.org/10.1007/s12257-009-0037-8