Folia Microbiologica

, Volume 56, Issue 6, pp 555–561 | Cite as

Mutational biosynthesis of neomycin analogs by a mutant of neomycin-producing Streptomyces fradiae

  • Guanying Shi
  • Xingang Zhang
  • Lang Wu
  • Jin Xie
  • Ke Tao
  • Taiping HouEmail author


Neomycin, produced by Streptomyces fradiae, has been widely used for the treatment of bacterial infections in clinical and agricultural applications. In this study, a neomycin nonproducing mutant of S. fradiae was obtained by gene disruption technique for mutational biosynthesis. A crucial gene neoC (neo7) which encodes 2-deoxystreptamine (2-DOS) synthases was disrupted. The mutant could resume producing neomycin in the presence of 2-DOS. Salen derivatives of 2-DOS were synthesized and individually added to cultures of the mutant. Antibacterial activity of the mutasynthesis products against Staphylococcus aureus and four plant pathogenic bacteria (Pseudomonas solanacarum, Erwinia carotovora, Xanthomonas oryzae, and Xanthomonas campestris) was detected quantitatively by Oxford cup method. It is suggested that all 2-DOS derivatives were incorporated by the mutant into new active neomycin analogs except for 2-DOS derivative 2d ((1R,2r,3S,4R,6S)-4,6-bis((E)-3,5-di-tert-butyl-2-hydroxybenzylideneamino)cyclohexane-1,2,3-triol). Neomycin analogs produced by feeding 2-DOS derivative 2a ((1R,2r,3S,4R,6S)-4,6-bis((E)-2 hydroxybenzylideneamino)cyclohexane-1,2,3-triol) to cultures of the mutant displayed a similar antibacterial activity with neomycin produced by wild strain.


Neomycin Fermentation Product Wild Strain Biosynthetic Gene Cluster Aminoglycoside Antibiotic 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This project was supported by grants from the National Natural Science Foundation of China (no. 30800034), the Science and Technology Department 11th Five-year Plan, State Science and Technology support projects (no. 2006BAE01A01-14), and the National High Technology Research and Development Program of China (863 Program, No. 2009AA032903). The authors also thank the Analytical Detective Center and Pharmaceutical School of Sichuan University for the NMR spectroscopic analysis.


  1. Brnakova Z, Godany A, Timko J (2009) Characterization and disruption of exonuclease genes from Streptomyces aureofaciens B96 and S. coelicolor A3(2). Folia Microbiol 54:97–104CrossRefGoogle Scholar
  2. Carlson H, Douglas H (1948) Screening methods for determining antibiotic activity of higher plants. J Bacteriol 55:235–240Google Scholar
  3. Cui WH, Quan X, Tao K, Teng Y, Liu Y, Shi GY, Hou TP (2009) Mechanism of action of neomycin on Erwinia carotovora subsp. carotovora. Pestic Biochem Phys 95:85–89CrossRefGoogle Scholar
  4. He Y, Tao K, Huang X, Cui WH, Zhang XG, Hou TP (2008) Control efficacy of neomycin against several bacterial plant diseases and its acute toxicity. Agrochemicals 47:297–299Google Scholar
  5. Hu GF (1998) Neomycin inhibits angiogenin-induced angiogensis. Proc Natl Acad Sci U S A 95:9791–9795PubMedCrossRefGoogle Scholar
  6. Hu GF (2001) Neomycin inhibits the angiogenic activity of fibroblast and epidermal growth factors. Biochem Biophys Res Commun 287:870–874PubMedCrossRefGoogle Scholar
  7. Huang FL, Haydock SF, Mireoenko T, Spiteller D, Li YY, Spencer JB (2005) The neomycin biosynthetic gene cluster of Streptomyces fradiae NCIMB 8233: characterisation of an aminotransferase involved in the formation of 2-deoxystreptamine. Org Biomol Chem 3:1410–1419PubMedCrossRefGoogle Scholar
  8. Kahlmeter G, Dahlager JI (1984) Aminoglycoside toxicity—a review of clinical studies published between 1975 and 1982. J Antimicrob Chemother 13:9–22PubMedGoogle Scholar
  9. Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. The John Innes Foundation, NorwichGoogle Scholar
  10. Koberska M, Kopecky J, Olsovska J, Jelinkova M, Ulanova D, Man P, Flieger M, Janata J (2008) Sequence analysis and heterologous expression of the lincomycin biosynthetic cluster of the type strain Streptomyces lincolnensis ATCC 25466. Folia Microbiol 53:395–401CrossRefGoogle Scholar
  11. Litovchick A, Evdokimov AG, Lapidot A (1999) Arginine-aminoglycoside conjugates that bind to HIV transactivation responsive element RNA in vitro. FEBS Lett 445:73–79PubMedCrossRefGoogle Scholar
  12. Litovchick A, Lapidot A, Eisenstein M, Kalinkovich A, Borkow G (2001) Neomycin B-arginine conjugate, a novel HIV-1 tat antagonist: synthesis and anti-HIV activities. Biochemistry 40:15612–15623PubMedCrossRefGoogle Scholar
  13. Llewellyn NM, Spencer JB (2006) Biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics. Nat Prod Rep 23:864–874PubMedCrossRefGoogle Scholar
  14. Mehta R, Champney WS (2002) 30S Ribosomal subunit assembly is a target for inhibition by aminoglycosides in Escherichia coli. Antimicrob Agents Chemother 46:1546–1549PubMedCrossRefGoogle Scholar
  15. Mehta R, Champney WS (2003) Neomycin and paromomycin inhibit 30S ribosomal subunit assembly in Staphylococcus aureus. Curr Microbiol 47:237–243PubMedCrossRefGoogle Scholar
  16. Myronovskyy M, Ostash B, Ostash I, Fedorenko V (2009) A gene cloning system for the siomycin producer Streptomyces sioyaensis NRRL-B5408. Folia Microbiol 54:91–96CrossRefGoogle Scholar
  17. Neu HC (1992) The crisis in antibiotic resistance. Science 257:1064–1073PubMedCrossRefGoogle Scholar
  18. Neu HC (1994) Antimicrobial chemotherapy, 1934–1994. Antimicrob Infect Dis Newsl 13:1–8CrossRefGoogle Scholar
  19. Pearce CJR, Rinehart KL Jr (1981) Antibiotics. Springer, Berlin, pp 74–100Google Scholar
  20. Rinehart KL Jr (1977) Mutasynthesis of new antibiotics. Pure Appl Chem 49:1361–1384CrossRefGoogle Scholar
  21. Rinehart KL Jr (1980) Biosynthesis and mutasynthesis of aminocyclitol antibiotics. Aminocyclitol Antibiotics 19:335–370CrossRefGoogle Scholar
  22. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor, New YorkGoogle Scholar
  23. Schroeder R, Waldsich C, Wank H (2000) Modulation of RNA function by aminoglycoside antibiotics. J EMBO 19:1–9CrossRefGoogle Scholar
  24. Shier WT, Rinehart KL Jr, Gottlieb D (1969) Preparation of four new antibiotics from a mutant of Streptomyces fradiae. Proc Natl Acad Sci 63:198–204PubMedCrossRefGoogle Scholar
  25. Subba B, Kharel MK, Lee HC, Liou K, Kim BG, Sohng JK (2005) The ribostamycin biosynthetic gene cluster in Streptomyces ribosidificus: comparison with butirosin biosynthesis. Mol Cells 20:90–96PubMedGoogle Scholar
  26. Testa RT, Wagman GH, Daniels PJ, Weinstein MJ (1974) Mutamicins; biosynthetically created new sisomicin analogs. J Antibiot 27:917–921PubMedGoogle Scholar
  27. Tok JBH, Dunn LJ, Des Jean RC (2001) Binding of dimeric aminoglycosides to the HIV-1 rev responsive element (RRE) RNA construct. Bioorg Med Chem Lett 11:1127–1131PubMedCrossRefGoogle Scholar
  28. Waksman SA, Lechevalier HA (1949) Neomycin, a new antibiotic active against streptomycin-resistant bacteria, including tuberculosis organisms. Science 109:305–307PubMedCrossRefGoogle Scholar
  29. Yanai K, Murakami T (2004) The kanamycin biosynthetic gene cluster from Streptomyces kanamyceticus. J Antibiotics 57:351–354Google Scholar

Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2011

Authors and Affiliations

  • Guanying Shi
    • 1
    • 2
  • Xingang Zhang
    • 1
    • 2
  • Lang Wu
    • 1
    • 2
  • Jin Xie
    • 2
  • Ke Tao
    • 2
  • Taiping Hou
    • 1
    • 2
    Email author
  1. 1.Key Laboratory of Bio-Resource and Eco-Environment of Ministry of EducationSichuan UniversityChengduPeople’s Republic of China
  2. 2.College of Life SciencesSichuan UniversityChengduPeople’s Republic of China

Personalised recommendations