Abstract
Ramoplanin is a lipopeptide antibiotic active against multi-drug-resistant, Gram-positive pathogens. Structurally, it contains a di-mannose moiety attached to the peptide core at Hpg11. The biosynthetic gene cluster of ramoplanin has already been reported and the assembly of the depsipeptide has been elucidated but the mechanism of transferring sugar moiety to the peptide core remains unclear. Sequence analysis of the biosynthetic gene cluster indicated ramo-orf29 was a mannosyltransferase candidate. To investigate the involvement of ramo-orf29 in ramoplanin biosynthesis, gene inactivation and complementation have been conducted in Actinoplanes sp. ATCC 33076 by homologous recombination. Metabolite analysis revealed that the ramo-orf29 inactivated mutant produced no ramoplanin but the ramoplanin aglycone. Thus, ramo-orf29 codes for the mannosyltransferase in the ramoplanin biosynthesis pathway. This lays the foundation for further exploitation of the ramoplanin mannosyltransferase and aglycone in combinatorial biosynthesis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bou G (2007) Minimum inhibitory concentration (MIC) analysis and susceptibility testing of MRSA. Methods Mol Biol 391:29
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. J Antibiot 37(4):309
Ciabatti R, Cavalleri B (1996) Aglycons of A/16686 antibiotics. US Patent 5,491,128
Hu Y, Helm JS, Chen L, Ye XY, Walker S (2003) Ramoplanin inhibits bacterial transglycosylases by binding as a dimer to lipid II. J Am Chem Soc 125(29):8736–8737
Jiang W, Wanner J, Lee RJ, Bounaud PY, Boger DL (2003) Total synthesis of the ramoplanin A2 and ramoplanose aglycon. J Am Chem Soc 125(7):1877–1887
Jones RN, Barry AL (1989) In vitro evaluation of ramoplanin (A16686 or MDL62198). A new depsipeptide complex for potential topical use. Diagn Microbiol Infect Dis 12(3):279–282
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical streptomyces genetics. John Innes Foundation, Norwich
Li TL, Huang F, Haydock SF, Mironenko T, Leadlay PF, Spencer JB (2004) Biosynthetic gene cluster of the glycopeptide antibiotic teicoplanin: characterization of two glycosyltransferases and the key acyltransferase. Chem Biol 11(1):107–119
Lo MC, Men H, Branstrom A, Helm J, Yao N, Goldman R, Walker S (2000) A new mechanism of action proposed for ramoplanin. J Am Chem Soc 122:3540–3541
Losey HC, Jiang J, Biggins JB, Oberthür M, Ye XY, Dong SD, Kahne D, Thorson JS, Walsh CT (2002) Incorporation of glucose analogs by GtfE and GtfD from the vancomycin biosynthetic pathway to generate variant glycopeptides. Chem Biol 9(12):1305–1314
Luzhetskyy A, Fedoryshyn M, Dürr C, Taguchi T, Novikov V, Bechthold A (2005) Iteratively acting glycosyltransferases involved in the hexasaccharide biosynthesis of landomycin A. Chem Biol 12(7):725–729
Magarvey NA, Haltli B, He M, Greenstein M, Hucul JA (2006) Biosynthetic pathway for mannopeptimycins, lipoglycopeptide antibiotics active against drug-resistant gram-positive pathogens. Antimicrob Agents Chemother 50(6):2167–2177
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(4):261–284. doi:10.1002/bip.10296
Paknikar SS, Narayana S (2012) Newer Antibacterials in Therapy and Clinical Trials. N Am J Med Sci 4(11):537
Parenti F, Ciabatti R, Cavalleri B, Kettenring J (1990) Ramoplanin: a review of its discovery and its chemistry. Drugs Experi Clin Res 16(9):451
Sambrook J, Russell D (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York
Sosio M, Stinchi S, Beltrametti F, Lazzarini A, Donadio S (2003) The gene cluster for the biosynthesis of the glycopeptide antibiotic A40926 by Nonomuraea species. Chem Biol 10(6):541–549
Sosio M, Kloosterman H, Bianchi A, de Vreugd P, Dijkhuizen L, Donadio S (2004) Organization of the teicoplanin gene cluster in Actinoplanes teichomyceticus. Microbiology 150(1):95–102
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(2):449–476
Walsh CT, Chen H, Keating TA, Hubbard BK, Losey HC, Luo L, Marshall CG, Miller DA, Patel HM (2001) Tailoring enzymes that modify nonribosomal peptides during and after chain elongation on NRPS assembly lines. Curr Opin Chem Biol 5(5):525–534
Zhang C, Bitto E, Goff RD, Singh S, Bingman CA, Griffith BR, Albermann C, Phillips GN, Thorson JS (2008) Biochemical and structural insights of the early glycosylation steps in calicheamicin biosynthesis. Chem Biol 15(8):842–853
Acknowledgments
This work was supported by Grants from National Natural Science Foundation of China (81072557, 81172962, and 81202442), the Shanghai Rising-Star Program (11QB1406300) and the Ministry of Science and Technology of China (Grant No. 2012ZX09201101-008).
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Chen, JS., Wang, YX., Shao, L. et al. Functional identification of the gene encoding the enzyme involved in mannosylation in ramoplanin biosynthesis in Actinoplanes sp.. Biotechnol Lett 35, 1501–1508 (2013). https://doi.org/10.1007/s10529-013-1233-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-013-1233-3