Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Isolation and Identification of a Bacillus sp. producing ${\alpha}$-glucosidase Inhibitor 1-deoxynojirimycin

알파글루코시다아제 저해제 1-deoxynojirimycin을 생산하는 Bacillus 균주의 분리 및 동정

  • 김현수 (수원대학교 생명과학과) ;
  • 이재연 ((주)바이오토피아 부설생명과학연구소) ;
  • 황교열 ((주)바이오토피아 부설생명과학연구소) ;
  • 조용석 ((주)바이오토피아 부설생명과학연구소) ;
  • 박영식 ((주)바이오토피아 부설생명과학연구소) ;
  • 강경돈 ((주)바이오토피아 부설생명과학연구소) ;
  • 성수일 ((주)바이오토피아 부설생명과학연구소)
  • Received : 2011.01.05
  • Accepted : 2011.02.19
  • Published : 2011.03.28
Download PDF
⟨ Previous Next ⟩

Abstract

Thirty Streptomyces sp. and 200 Bacillus sp. isolated from Korean soils and traditional foods were screened for their abilities to inhibit ${\alpha}$-glucosidase and produce 1-deoxynojirimycin (DNJ). This screening identified a Bacillus sp. bacterium that strongly inhibited ${\alpha}$-glucosidase and produced high levels of DNJ from Chungkookjang, a Korean traditional food. The bacterium was characterized in terms of its biochemical and molecular biological properties such as sugar utilization, cellular quinone composition, cell wall fatty acid composition, and 16S rDNA sequence. In addition, scanning electron microscopy was used to visualize the morphology of the bacterium. These analyses identified the bacterium as B. subtilis, a bacterium with Generally Recognized as Safe (GRAS) status. The selected strain was named B. subtilis MORI.

토양으로부터 분리한 Streptomyces sp. 30여 균주들과 한국전통식품에서 분리한 Bacillus sp. 200여 균주들로부터 ${\alpha}$-glucosidase 의 활성을 저해하고 동시에 DNJ를 생산하는 유용균주를 선발하였다. 실험결과 한국전통식품인 청국장으로부터 ${\alpha}$-glucosidase 저해능이 높고 DNJ 생산능이 우수한 한개의 균주를 선발하였다. 이 균주의 동정을 위하여 API kit에 의한 균의 당 이용능 분석, HPLC와 GC에 의한 균체의 quinone 및 지방산 분석 등과 함께 균의 16S rDNA 염기서열을 분석하였으며 주사전자현미경에 의해 균주의 형태적 특성을 관찰하였다. 그 결과 본 연구를 통해 선발한 균주는 건강기능성식품 개발 등에 적용할 수 있는 GRAS에 속하는 균주임을 확인하여 B. subtilis MORI로 명명하였다.

Keywords

References

  1. Asano, N., A. Kato, K. Oseki, H. Kizu, and K. Matsui. 1995. Calystegins of Physalis alkekengi var. francheti (Solanaceae). Structure determination and their glycosidase inhibitory activities. Eur. J. Biochem. 229: 369-376. https://doi.org/10.1111/j.1432-1033.1995.0369k.x
  2. Asano, N., R. J. Nash, R. J. Molyneux, and G. W. Fleet. 2000. Sugar-mimic glycosidase inhibitors: natural occurrence, biological activity and prospects for therapeutic application. Tetrahedron 11: 1645-1680. https://doi.org/10.1016/S0957-4166(00)00113-0
  3. Asano, N., T. Yamashita, K. Yasuda, K. Ikeda, H. Kizu, Y. Kameda, A. Kato, R. J. Nash, H. S. Lee, and K. S. Ryu. 2001. Polyhydroxylated alkaloids isolated from mulberry trees(Morus alba L.) and silkworms(Bombyx mori L.). J. Agric. Food. Chem. 49: 4208-4213. https://doi.org/10.1021/jf010567e
  4. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248- 254. https://doi.org/10.1016/0003-2697(76)90527-3
  5. Davis, B. 1964. Disc electrophoresis-II. Method and application to human serum proteins. Ann. New York Acad. Sci. 121: 404-427.
  6. Inouye, S., T. Tsuruoka, T. Ito, and T. Niida. 1966. The structure of nojirimycin, a piperidinose sugar antibiotic. J. Antibiot. Ser. A. 19: 288-292.
  7. Inouye, S., T. Tsuruoka, T. Ito, and T. Niida. 1968. Structure and synthesis of nojirimycin. Tetrahedron. 24: 2125-2144. https://doi.org/10.1016/0040-4020(68)88115-3
  8. Ishida, N., K. Kumagai, T. Niida, T. Tsuruoka, and H. Yumoto. 1967. A new antibiotic, nojirimycin. II. Isolation, characterization and biological activity. J. Antibiot. Ser. A. 20: 66-71.
  9. Kim, J. W., S. U. Kim, H. S. Lee, I. S. Kim, M. Y. Ahn, and K. S. Ryu. 2003. Determination of 1-deoxynojirimycin in Morus alba L. leaves using derivatization with 9- fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography. J. Chromatogr. A. 1002: 93-99. https://doi.org/10.1016/S0021-9673(03)00728-3
  10. Murao, S. and S. Miyata. 1980. Isolation and characterization of a new trehalase inhibitor, S-GI. Agric. Biol. Chem. 44: 219-221. https://doi.org/10.1271/bbb1961.44.219
  11. Olden, K., P. Breton, K. Grzegorzewski, Y. Yasuda, B. L. Gause, J. K. Cha, V. A. Miles, S. A. Newtown, and S. L. White. 1991. The potential importance of swainsonine in therapy for cancers and immunology. Pharmacol. Ther. 50: 285-290. https://doi.org/10.1016/0163-7258(91)90046-O
  12. Schmidt, D. D., W. Frommer, L. Müller, and E. Truscheit. 1979. Glucosidase-inhibtoren aus Bazillen. Naturwissenshaften. 66: 584-585.
  13. Shin, Y. K, J. S. Lee, C. O. Chun, H. J. Kim, and Y. H. Park. 1996. Isoprenoid quinone profiles of the Leclercia adecarboxylata KCTC 1036. J. Microbiol. Biotech. 6: 68- 69.
  14. Taylor, D. L., P. Sunkara, P. S. Liu, M. S. Kang, T. L. Bowlin, and A. S. Tyms. 1991. 6-O-Butanoylcastanospermine (MDL 28,574) inhibits glycoprotein processing and the growth of HIVs. AIDS 5: 693-698. https://doi.org/10.1097/00002030-199106000-00008
  15. Watson, A. A., G. W. Fleet, N. Asano, R. J. Molyneux, and R. J. Nash. 2001. Polyhydroxylated alkaloids-natural occurrence and therapeutic applications. Phytochemistry 56: 265-295. https://doi.org/10.1016/S0031-9422(00)00451-9
  16. Yagi, M., T. Kouno, Y. Aoyagi, and H. Murai. 1976. The structure of moranoline, a piperidine alkaloid from Morus species. Nippon Nogeikagaku Kaishi. 50: 571-572. https://doi.org/10.1271/nogeikagaku1924.50.11_571