Isolation and Characterization of Kasugamycin Biosynthetic Genes from Streptomyces kasugaensis KACC 20262

  • JO YOU-YOUNG (Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University) ;
  • LIU JING (Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University) ;
  • JIN YING-YU (Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University) ;
  • YANG YOUNG-YELL (Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University) ;
  • SUH JOO-WON (Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University)
  • Published : 2005.06.01

Abstract

The biosynthetic gene cluster for the aminoglycoside antibiotic kasugamycin was isolated and characterized from the kasugamycin producing strain, Streptomyces kasugaensis KACC 20262. By screening a fosmid library using kasA, the gene encoding aminotransferase, we isolated a 22 kb DNA fragment. The fragment contained seventeen complete open reading frames (ORFs); one of these ORFs, kasD, was identified as the gene for dNDP-glucose 4,6-dehydratase, which catalyzes the conversion of dNDP-glucose to 4-keto-6-deoxy-dNDP-glucose. The enzyme showed a broad spectrum of substrate specificity. In addition, ksR was overexpressed in E. coli BL21 and proved to be a self-resistance gene against kasugamycin. These findings suggest that the isolated gene cluster is highly likely responsible for the biosynthesis of kasugamycin.

Keywords

References

  1. Altschul, S. F. and D. J. Lipman. 1990. Protein database searches for multiple alignments. Proc. Natl. Acad. Sci. USA 87: 5509-5513
  2. Benveniste, R. and J. Davies. 1973. Aminoglycoside antibiotic inactivating enzymes in actinomycetes similar to those present in clinical isolates of antibiotic-resistant bacteria. Proc. Nat. Acad. Sci. USA 70: 2276-2280
  3. Bibb, M. J., P. R. Findlay, and M. W. Johnson, 1984. The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene 30: 157-166 https://doi.org/10.1016/0378-1119(84)90116-1
  4. Cho, J. and R. R. Rando. 1999. Specificity in the binding of aminoglycosides to HIV-RRE RNA. Biochemistry 38: 8548-8554 https://doi.org/10.1021/bi990273a
  5. Higgins, D. G. and P. M. Sharp. 1968. CLUSTAL: A package for performing multiple sequence alignments on a microcomputer. Gene 73: 237-244 https://doi.org/10.1016/0378-1119(88)90330-7
  6. Ichikawa, T. and H. Umezawa. 1967. Kasugamycin in Pseudomonas infections of the urinary tract. J. Urol. 97: 917-925. https://doi.org/10.1016/S0022-5347(17)63149-5
  7. Ikeno, S., D. Aoki, K. Sato, M. Hamada, M. Hori, and K. S. Tsuchiya. 2002. kasT gene of Streptomyces kasugaensis M3380M1 encodes a DNA-binding protein which binds to intergenic region of kasU-kasJ in the kasugamycin biosynthesis gene cluster. J. Antibiot. 55: 1053-1062 https://doi.org/10.7164/antibiotics.55.1053
  8. Ikeno, S., T. Tsuji, K. Higashide, and N. Kinoshita. 1998. A 7.6 kb DNA region from Streptomyces kasugaensis M338M1 includes some genes responsible for kasugamycin biosynthesis. J. Antibiot. 51: 341-351 https://doi.org/10.7164/antibiotics.51.341
  9. Jo, Y. Y., S. H. Kim, Y. Y. Yang, C. M. Kang, J. K. Sohng, and J. W. Suh. 2003. Functional analysis of spectinomycin biosynthetic genes from Streptomyces spectabilis ATCC 27741. J. Microbiol. Biotechnol. 13: 906-911
  10. Jung, Y. G., S. H. Kang, C. G.. Hyun, Y. Y. Yang, C. M. Kang, and J. W. Suh. 2003. Isolation and characterization of bluensomycin biosynthetic genes trom Streptomyces bluensis. FEMS Microbiol. Lett. 219: 285-289 https://doi.org/10.1016/S0378-1097(03)00019-3
  11. Kennington, A. S., C. R. Hill, J. Craig, C. Bogardus, I. Raz, H. K. Ortmeyer, B. C. Hansen, G. Romero, and J. Larner. 1990. Low urinary chiro-inositol excretion in non-insulindependent diabetes mellitus. N Engl. J. Med. 323: 373-378 https://doi.org/10.1056/NEJM199008093230603
  12. Kharel, M. K., S. Bimala, H. C. Lee, K. K. Liou, J. S. Woo, D. H. Kim, and J. K. Sohng. 2003. Identification of 2-deoxy-scyllo-inosose synthase in aminoglycoside producer Streptomyces. J. Microbiol. Biotechnol. 13: 828-831
  13. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0
  14. Lee, H. C., J. Y. Sohge, H. J. Kim, D. Y. Nam, C. N. Seong, J. M. Han, and J. C. Yoo. 2004. Cloning, expression, and biochemical characterization of dTDP-glucose 4,6-dehydratase gene (gerE) trom Streptomyces sp. GERI-155. J. Microbiol. Biotechnol. 14: 576-583
  15. Litovhick, A., A. G. Evdokimov, and A. Lapidot. 2000. Aminoglycoside-arginine conjugates that bind TAR RNA: Synthesis, characterization, and antiviral activity. Biochemistry 39: 2838-2852 https://doi.org/10.1021/bi9917885
  16. Neuwald, A. F. and D. Landsman. 1997. GCN5-related histone N-acetyltransferases belong to a diverse superfamily that includes the yeast SPT 10 protein. Trends Biochem. Sci. 22: 154-155 https://doi.org/10.1016/S0968-0004(97)01034-7
  17. Ohta, T., T. Dairi, E. Hashimoto, and M. Hasegawa. 1993. Use of a heterologous gene for molecular breeding of actinomycetes producing structurally related antibiotics: Self-defense genes of producers of fortimicin-A (astromicin)group antibiotics. Actinomycetologica 7: 145-155 https://doi.org/10.3209/saj.7_145
  18. Piepersberg, W. 1995. Streptomycin and related aminoglycosides, pp. 531-570. In Vining, L. and Stuttard, C. (eds.), Biochemistry and Genetics of Antibiotic Biosynthesis. Butterworth-Heinemann, Boston
  19. Pipersberg, W. 1997. Molecular biology, biochemistry, and fermentation of aminoglycoside antibiotics, pp. 81-163. In Strohl, W. R. (eds.). Biotechnology of Antibiotics. Marcel-Dekker Inc., New York, U.S.A.
  20. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, U.S.A.
  21. Schoner, G., L. F. Ellis, and B. E. Schoner. 1985. Isolation and purification of protein granules from Escherichia coli overexproducing biosynthetic human insulin proteins. Bio/Technology 3: 151- 154 https://doi.org/10.1038/nbt0285-151
  22. Suhara, Y., F. Sasaki, G. Koyama, K. Maeda, H. Umezawa, and M. Ono. 1972. The total synthesis of kasugamycin. J. Am. Chem. Soc. 94: 6501-6507 https://doi.org/10.1021/ja00773a038
  23. Tanaka, N., T. Nishimura, H. Yamaguchi, C. Yamamoto, Y. Yoshida, K. Sashikata, and H. Umezawa. 1965. Mechanism of action of kasugamycin. J. Antibiot. 18: 139-144
  24. Tanaka, N., Y. Yoshida, K. Sashikata, H. Yamaguchi, and H. Umezawa. 1966. Inhibition of polypeptide synthesis by kasugamycin, an aminoglycosidic. J. Antibiot. 19:65-68
  25. Umezawa, H. and I. R. Hopper. 1982. Aminoglycoside Antibiotics. pp. 171-176. Springer-Verlag Berlin Heidelberg New York
  26. Umezawa, H., Y. Okami, T. Hashimoto, Y. Suhara, M. Hamada, and T. Takeuchi. 1965. A new antibiotic, kasugamycin. J. Antibiot. Ser. A 18: 101-103
  27. Vara, J. A. and C. R. Hutchinson. 1988. Purification of thymidine-diphospho-D-glucose 4,6-dehydratase from an erythromycin producing strain of Saccharopolyspora erythraea by high resolution liquid chromatography. J. Biol. Chem. 263: 14992-14995
  28. Zhao, X. Q., K. R. Kim, L. W. Sang, S. H. Kang, Y. Y. Yang, and J. W. Suh. 2005. Genetic organization of a 50-kb gene cluster isolated from Streptomyces kanamyceticus for kanamycin biosynthesis and characterization of kanamycin acetyltransferase. J. Microbiol. Biotechnol. 15: 346-353