Heterologous production of daptomycin in Streptomyces lividans

  • Julia Penn
  • Xiang Li
  • Andrew Whiting
  • Mohammed Latif
  • Trevor Gibson
  • Christopher J. Silva
  • Paul Brian
  • Julian Davies
  • Vivian Miao
  • Stephen K. Wrigley
  • Richard H. Baltz
Original Paper

Abstract

Daptomycin and the A21978C antibiotic complex are lipopeptides produced by Streptomyces roseosporus and also in recombinant Streptomyces lividans TK23 and TK64 strains, when a 128 kbp region of cloned S. roseosporus DNA containing the daptomycin gene cluster is inserted site-specifically in the ϕC31 attB site. A21978C fermentation yields were initially much lower in S. lividans than in S. roseosporus, and detection was complicated by the production of host metabolites. However A21978C production in S. lividans was improved by deletion of genes encoding the production of actinorhodin and by medium optimization to control the chemical form of the calcium dependent antibiotic (CDA). This latter compound has not previously been chemically characterized as a S. lividans product. Adding phosphate to a defined fermentation medium resulted in formation of only the phosphorylated forms of CDA, which were well separated from A21978C on chromatographic analysis. Adjusting the level of phosphate in the medium led to an improvement in A21978C yield from 20 to 55 mg/l.

Keywords

Daptomycin A21978C Calcium-dependent antibiotic (CDA) Actinorhodin Streptomyces lividans 

References

  1. 1.
    Bibb MJ, Janssen GR, Ward JM (1985) Cloning and analysis of the promoter region of the erythromycin resistance gene (ermE) of Streptomyces erythraeus. Gene 38:215–226CrossRefPubMedGoogle Scholar
  2. 2.
    Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp . Gene 116:43–49CrossRefPubMedGoogle Scholar
  3. 3.
    Chouayekh H, Virolle M-J (2002) The polyphosphate kinase plays a negative role in the control of antibiotic production in Streptomyces lividans. Mol Microbiol 43:919–930CrossRefPubMedGoogle Scholar
  4. 4.
    Debono M, Barnhart M, Carrell CB, Hoffman JA, Occolowitz JL, Abbott BJ, Fukuda DS, Hamill RL (1987) A21978C, a complex of new acidic peptide antibiotics: isolation, chemistry, and mass spectral structure elucidation. J Antibiot 40:761–777PubMedGoogle Scholar
  5. 5.
    Debono M, Abbott BJ, Molloy RM, Fukuda DS, Hunt AH, Daupert VM, Counter FT, Ott JL, Carrell CB, Howard LC, Boeck LD, Hamill RL (1988) Enzymatic and chemical modifications of lipopetide antibiotic A21978C: the synthesis and evaluation of daptomycin (LY146032). J Antibiot 41:1093–1105PubMedGoogle Scholar
  6. 6.
    Eisenstein BI (2004) Lipopeptides, focusing on daptomycin, for the treatment of Gram-positive infections. Expert Opin Investig Drugs 13:1159–1169CrossRefPubMedGoogle Scholar
  7. 7.
    Gorst-Allman CP, Rudd BAM, Chang C, Floss HG (1981) Biosynthesis of actinorhodin. Determination of the point of dimerization. J Org Chem 46:455–456CrossRefGoogle Scholar
  8. 8.
    Hojati Z, Milne C, Harvey B, Gordon L, Borg M, Flett F, Wilkinson B, Sidebottom PJ, Rudd BAM, Hayes MA, Smith CP, Micklefield J (2002) Structure, biosynthetic origin, and engineered biosynthesis of calcium-dependent antibiotics from Streptomyces coelicolor. Chem Biol 9:1–13CrossRefPubMedGoogle Scholar
  9. 9.
    Hu H, Zhang Q, Ochi K (2002) Activation of antibiotic biosynthesis by specified mutations in the rpoB gene (encoding the RNA polymerase β subunit) of Streptomyces lividans. J Bacteriol 184:3984–3991CrossRefPubMedGoogle Scholar
  10. 10.
    Huber FM, Pieper RL, Tietz AJ (1988) The formation of daptomycin by supplying decanoic acid to Streptomyces roseosporus cultures producing the antibiotic complex A21978C. J Biotechnol 7:283–292CrossRefGoogle Scholar
  11. 11.
    Jung D, Rozek A, Okon M, Hancock REW (2004) Structural transitions as determinants of the action of the calcium-dependent antibiotic daptomycin. Chem Biol 11:949–957CrossRefPubMedGoogle Scholar
  12. 12.
    Keiser T, Bibb M, Buttner MJ, Chater KF, Hopwood D (2000) Practical Streptomycete genetics. John Innes Foundation, NorwichGoogle Scholar
  13. 13.
    Kempter C, Kaiser D, Haag S, Nicholson G, Gnau V, Walk T, Gierling KH, Decker H, Zahner H, Jung G, Metzger JW (1997) CDA: calcium-dependent peptide antibiotics from Streptomyces coelicolor A3(2) containing unusual residues. Angew Chem Int Ed Engl 36:498–501CrossRefGoogle Scholar
  14. 14.
    LaPlante KL, Rybak MJ (2004) Daptomycin – a novel antibiotic against Gram-positive pathogens. Expert Opin Pharmacother 5:2321–2331CrossRefPubMedGoogle Scholar
  15. 15.
    Malpartida F, Hopwood DA (1984) Molecular cloning of the whole biosynthetic pathway of a Streptomyces antibiotic and its expression in a heterologous host. Nature 309: 462–464CrossRefPubMedGoogle Scholar
  16. 16.
    Malpartida F, Hopwood DA (1986) Physical and genetic characterisation of the gene cluster for the antibiotic actinorhodin in Streptomyces coelicolor A3(2). Mol Gen Genet 205:66–73PubMedCrossRefGoogle Scholar
  17. 17.
    Miao V, Coeffet-LeGal M-F, Brian P, Brost R, Penn J, Whiting A, Martin S, Ford R, Parr I, Bouchard M, Silva CJ, Wrigley SK, Baltz RH (2005) Daptomycin biosynthesis in Streptomyces roseosporus: cloning and analysis of the gene cluster and revision of peptide stereochemistry. Microbiology 151:1507–1523CrossRefPubMedGoogle Scholar
  18. 18.
    Raja A, LaBonte J, Lebbos J, Kirkpatrick P (2003) Fresh from the pipeline: daptomycin. Nat Rev Drug Disc 2:943–944CrossRefGoogle Scholar
  19. 19.
    Shima J, Hesketh A, Okamoto S, Kawamoto S, Ochi K (1996) Induction of actinorhodin production by rpsL (encoding ribosomal protein S12) mutations that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2). J Bacteriol 178:7276–7284PubMedGoogle Scholar
  20. 20.
    Silverman JA, Perlmutter NG, Shapiro HM (2003) Correlation of daptomycin bactericidal activity and membrane depolarization in Staphylococcus aureus. Antimicrob Agent Chemother 47:2538–2544CrossRefGoogle Scholar
  21. 21.
    Zeeck A, Christiansen P (1969). Konformation and Konfiguration des Actinorhodins. Liebigs Ann Chem 724:172–182CrossRefGoogle Scholar

Copyright information

© Society for Industrial Microbiology 2005

Authors and Affiliations

  • Julia Penn
    • 1
  • Xiang Li
    • 2
  • Andrew Whiting
    • 1
  • Mohammed Latif
    • 1
  • Trevor Gibson
    • 1
  • Christopher J. Silva
    • 2
  • Paul Brian
    • 2
  • Julian Davies
    • 2
    • 3
  • Vivian Miao
    • 2
  • Stephen K. Wrigley
    • 1
  • Richard H. Baltz
    • 2
  1. 1.Cubist Pharmaceuticals (UK) LtdSloughUK
  2. 2.Cubist Pharmaceuticals, IncLexingtonUSA
  3. 3.Dept of Microbiology and ImmunologyUniversity of British ColumbiaVancouverCanada

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