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Characterisation of actI-homologous DNA encoding polyketide synthase genes from the monensin producer Streptomyces cinnamonensis

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Summary

Cloned DNA encoding polyketide synthase (PKS) genes from one Streptomyces species was previously shown to serve as a useful hybridisation probe for the isolation of other PKS gene clusters from the same or different species. In this work, the actI and actIII genes, encoding components of the actinorhodin PKS of Streptomyces coelicolor, were used to identify and clone a region of homologous DNA from the monensin-producing organism S. cinnamonensis. A 4799 by fragment containing the S. cinnamonensis act-homologous DNA was sequenced. Five open reading frames (ORFs 1–5) were identified on one strand of this DNA. The five ORFs show high sequence similarities to ORFs that were previously identified in the granaticin, actinorhodin, tetracenomycin and whiE PKS gene clusters. This allowed the assignment of the following putative functions to these five ORFS : a heterodimeric β-ketoacyl synthase (ORF1 and ORF2), an acyl carrier protein (ORF3), a β-ketoacyl reductase (ORF5), and a bifunctional cyclase/dehydrase (ORF4). The ORFs are encoded in the order ORFl-ORF2-ORF3-ORF5-ORF4, and ORFs-1 and -2 show evidence for translational coupling. This act-homologous region therefore appears to encode a PKS gene cluster. A gene disruption experiment using the vector pGM 160, and other evidence, suggests that this cluster is not essential for monensin biosynthesis but rather is involved in the biosynthesis of a cryptic aromatic polyketide in S. cinnamonensis. An efficient plasmid transformation system for S. cinnamonensis has been established, using the multicopy plasmids pWOR120 and pWOR125.

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Author notes

  1. Teresa J. Arrowsmith

    Present address: SmithKline Beecham Pharmaceuticals, Clarendon Road, BN14 8QH, Worthing, West Sussex, UK

  2. Francisco Malpartida

    Present address: Centro Nacional de Biotecnologia, Serrano 115 bis, E-28006, Madrid, Spain

  3. David H. Sherman

    Present address: Institute for Advanced Studies in Biological Process Technology, and Department of Microbiology, University of Minnesota, 1479 Gortner Avenue, M-55108, St Paul, USA

Authors and Affiliations

  1. John Innes Institute, NR4 7UH, Colney Lane, Norwich, UK

    Teresa J. Arrowsmith, Francisco Malpartida, David H. Sherman & David A. Hopwood

  2. Institute of Organic Chemistry, University of Zürich, Winterthurerstrasse 190, CH-8057, Zürich, Switzerland

    Ashley Birch & John A. Robinson

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  1. Teresa J. Arrowsmith
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  2. Francisco Malpartida
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Communicated by H. Hennecke

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Arrowsmith, T.J., Malpartida, F., Sherman, D.H. et al. Characterisation of actI-homologous DNA encoding polyketide synthase genes from the monensin producer Streptomyces cinnamonensis . Molec. Gen. Genet. 234, 254–264 (1992). https://doi.org/10.1007/BF00283846

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