Skip to main content
Log in

Genetic manipulation revealing an unusual N-terminal region in a stand-alone non-ribosomal peptide synthetase involved in the biosynthesis of ramoplanins

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Ramoplanins produced by Actinoplanes are new structural class of lipopeptide and are currently in phase III clinical trials for the prevention of vancomycin-resistant enterococcal infections. The depsipeptide structures of ramoplanins are synthesized by non-ribosomal peptide synthetases (NRPS). Romo-orf17, a stand-alone NRPS, is responsible for the recruitment of Thr into the linear NRPS pathways for which the corresponding adenylation domain is absent. Here, systematical gene inactivation and complementation have been carried out in a Actinoplanes sp. using homologous recombination and site-specific integration methods. A hybrid gene coding for the N-terminal region of the stand-alone NRPS and the A-PCP domains of a heterologous NRPS restored production of ramoplanins. The results elucidate the unusual N-terminal region which is essential for the biosynthesis of ramoplanins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Butler MS (2008) Natural products to drugs: natural product-derived compounds in clinical trials. Nat Prod Rep 25:475–516

    Article  PubMed  CAS  Google Scholar 

  • Cavalleri B, Pagani H, Volpe G, Selva E, Parenti F (1984) A-16686, a new antibiotic from Actinoplanes. I. Fermentation, isolation and preliminary physico-chemical characteristics. Antibiotics 37:309–317

    Article  CAS  Google Scholar 

  • Cragg GM, Grothaus PG, Newman DJ (2009) Impact of natural products on developing new anti-cancer agents. Chem Rev 109:3012–3043

    Article  PubMed  CAS  Google Scholar 

  • Farnet CM, Zazopoulos E, Staffa A (2002) In: PCT Int Appl (Ecopia Biosciences Inc., Can.): Wo 0231155.p 212

  • Felnagle EA, Jackson EE, Chan YA, Podevels AM, Berti AD, McMahon MD, Thomas MG (2008) Non-ribosomal peptide synthetases involved in the production of medically relevant natural products. Mol Pharm 5:191–211

    Article  PubMed  CAS  Google Scholar 

  • Fischbach MA, Walsh CT (2006) Assembly-line enzymology for polyketide and non-ribosomal Peptide antibiotics: logic, machinery, and mechanisms. Chem Rev 106:3468–3496

    Article  PubMed  CAS  Google Scholar 

  • Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical streptomyces genetics. John Innes Foundation, Norwich

    Google Scholar 

  • McCafferty DG, Cudic P, Frankel BA, Barkallah S, Kruger RG, Li W (2002) Chemistry and biology of the ramoplanin family of peptide antibiotics. Biopolymers 66:261–284

    Article  PubMed  CAS  Google Scholar 

  • Parenti F, Ciabatti R, Cavalleri B, Kettenring J (1990) Ramoplanin: a review of its discovery and its chemistry. Drugs Exp Clin Res 16:451–455

    PubMed  CAS  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Strieker M, Tanović A, Marahiel MA (2010) Non-ribosomal peptide synthetases: structures and dynamics. Curr Opin Struct Biol 20:234–240

    Article  PubMed  CAS  Google Scholar 

  • Sundlov JA, Shi C, Wilson DJ, Aldrich CC, Gulick AM (2012) Structural and functional investigation of the intermolecular interaction between NRPS adenylation and carrier protein domains. Chem Biol 19:188–198

    Article  PubMed  CAS  Google Scholar 

  • Walker S, Chen L, Hu Y, Rew Y, Shin D, Boger DL (2005) Chemistry and biology of ramoplanin: a lipoglycodepsipeptide with potent antibiotic activity. Chem Rev 105:449–475

    Article  PubMed  CAS  Google Scholar 

  • Yin X, Zabriskie TM (2006) The enduracidin biosynthetic gene cluster from Streptomyces fungicidicus. Microbiology 152:2969–2983

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported in part by grants from the National Natural Science Foundation of China (81072557, 81172962 and 30801449), the Science and Technology Commission of Shanghai Municipality (11QB1406300) and the Ministry of Science and Technology of China (Grant No. 2009ZX09301-007).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Gong-Li Tang or Dai-Jie Chen.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pan, HX., Li, JA., Shao, L. et al. Genetic manipulation revealing an unusual N-terminal region in a stand-alone non-ribosomal peptide synthetase involved in the biosynthesis of ramoplanins. Biotechnol Lett 35, 107–114 (2013). https://doi.org/10.1007/s10529-012-1056-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-012-1056-7

Keywords

Navigation