HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplementary table Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Google Scholar Articles by Doekel, S. Articles by Brian, P. PubMed PubMed Citation Articles by Doekel, S. Articles by Brian, P. Agricola Articles by Doekel, S. Articles by Brian, P.

Non-ribosomal peptide synthetase module fusions to produce derivatives of daptomycin in Streptomyces roseosporus

Cubist Pharmaceuticals Inc., 65 Hayden Avenue, Lexington, MA 02421, USA

Correspondence
Richard H. Baltz
rbaltz{at}cubist.com

Genetic engineering has been applied to reprogramme non-ribosomal peptide synthetases (NRPSs) to produce novel antibiotics, but little is known about what determines the efficiency of production. We explored module exchanges at nucleotide sequences encoding interpeptide linkers in dptD, a gene encoding a di-modular NRPS subunit that incorporates 3-methylglutamic acid (3mGlu₁₂) and kynurenine (Kyn₁₃) into daptomycin. Mutations causing amino acid substitutions, deletions or insertions in the inter-module linker had no negative effects on lipopeptide yields. Hybrid DptD subunits were generated by fusing the 3mGlu₁₂ module to terminal modules from calcium-dependent antibiotic (CDA) or A54145 NRPSs, and recombinants produced daptomycin analogues with Trp₁₃ or Ile₁₃ at high efficiencies. A recombinant expressing DptD with a hybrid Kyn₁₃ module containing a di-domain from a D-Asn module caused the production of a new daptomycin analogue containing Asn₁₃.

A supplementary table of primers is available with the online version of this paper.