Pacidamycin biosynthesis: identification and heterologous expression of the first uridyl peptide antibiotic gene cluster.
The pacidamycins are antimicrobial nucleoside antibiotics produced by Streptomyces coeruleorubidus that inhibit translocase I, an essential bacterial enzyme yet to be clinically targeted. The novel pacidamycin scaffold is composed of a pseudopeptide backbone linked by a unique exocyclic enamide to an atypical 3'-deoxyuridine nucleoside. In addition, the peptidyl chain undergoes a double inversion caused by the incorporation of a diamino acid residue and a rare internal ureido moiety. The pacidamycin gene cluster was identified and sequenced, thereby providing the first example of a biosynthetic cluster for a member of the uridyl peptide family of antibiotics. Analysis of the 22 ORFs provided an insight into the biosynthesis of the unique structural features of the pacidamycins. Heterologous expression in Streptomyces lividans resulted in the production of pacidamycin D and the newly identified pacidamycin S, thus confirming the identity of the pacidamycin biosynthetic gene cluster. Identification of this cluster will enable the generation of new uridyl peptide antibiotics through combinatorial biosynthesis. The concise cluster will provide a useful model system through which to gain a fundamental understanding of the way in which nonribosomal peptide synthetases interact.
Rackham
Emma J
EJ
School of Chemistry, University of East Anglia, Earlham Road, Norwich NR4 7TJ, UK.
Grüschow
Sabine
S
Ragab
Amany E
AE
Dickens
Shilo
S
Goss
Rebecca J M
RJ
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D005810:Multigene Family; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction; D011741:Pyrimidine Nucleosides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D010455:Peptides; D011741:Pyrimidine Nucleosides; C059936:pacidamycin 1; D004247:DNA