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Genetic analysis of polyketide synthase and peptide synthetase genes in cyanobacteria as a mining tool for secondary metabolites

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

Abstract

Molecular screening using degenerate PCR to determine the presence of secondary metabolite genes in cyanobacteria was performed. This revealed 18 NRPS and 19 PKS genes in the 21 new cyanobacterial strains examined, representing three families of cyanobacteria (Nostocales, Chroococales and Oscillatoriales). A BLAST analysis shows that these genes have similarities to known cyanobacterial natural products. Analysis of the NRPS adenylation domain indicates the presence of novel features previously ascribed to both proteobacteria and cyanobacteria. Furthermore, binding-pocket predictions reveal diversity in the amino acids used during the biosynthesis of compounds. A similar analysis of the PKS ketosynthase domain shows significant structural diversity and their presence in both mixed modules with NRPS domains and individually as part of a PKS module. We have been able to classify the NRPS genes on the basis of their binding-pockets. Further, we show how this data can be used to begin to link structure to function by an analysis of the compounds Scyptolin A and Hofmannolin from Scytonema sp. PCC 7110.

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Acknowledgments

We would like to thank Helia Radianingtyas for the supply of most of the genomic DNA used here. We also thank the Overseas Research Student’s Award Scheme and The University of Sheffield’s PhD Scholarship Programme for provision of a PhD scholarship to MBL. This work was supported by the UK Engineering and Physical Science Research Council (EPSRC), and The Carbon Trust. PCW wishes to thank the EPSRC for an advanced research fellowship.

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  1. Martin E. Barrios-Llerena

    Present address: Marie Curie Excellence Team Pathogen Habitats, Institute of Immunology and Infection Research, University of Edinburgh, West Mains Road, Edinburgh, EH9 3JT, UK

Authors and Affiliations

  1. Biological and Environmental Systems Group, Department of Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield, S1 3JD, UK

    Martin E. Barrios-Llerena & Phillip C. Wright

  2. Metabolic Engineering and Fermentation Group, Ocean Nutrition Canada, 101 Research Drive, Dartmouth, NS, B2Y 4T6, Canada

    Adam M. Burja

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10295_2007_216_MOESM1_ESM.doc

Figure 1. Alignment of the active sites of type I ketosynthase (KS) domains with distinct functions. Conserved residues are highlighted in black. Residues conserved within each functional group are highlighted in gray. (A) Representatives of KS domains specific for amino acid starter units in mixed or hybrid systems. Sequences included here are BarE from Lyngbya majuscula (accession No. AAN32979.1), NosB from Nostoc sp. (accession No. AF204805), EposB from Sorangium cellulosum (accession No. AF217189), MtaD from Stigmatella aurantiaca (accession No. AAF19812), and BlmVIII from Streptomyces verticillus (accession No. AAG02357). (B) Representatives of type I KS domain, sequences analysed included MycD and E from Microcystis aeruginosa (accession No. AF183408), JamE from Lyngbya majuscula (accession No. AY522504), CurA, I and F from Lyngbya majuscula (accession No. AY652953), EposA and D from Sorangium cellulosum (accession No. AF217189), and LnmI from Streptomyces atroolivaceus (accession No. AF484556). The Type I fatty acid synthase KS domain sequence from Escherichia coli (accession No. 6573501) is also given as a comparison and the consensus sequence is shown. (DOC 41 kb)

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Barrios-Llerena, M.E., Burja, A.M. & Wright, P.C. Genetic analysis of polyketide synthase and peptide synthetase genes in cyanobacteria as a mining tool for secondary metabolites. J Ind Microbiol Biotechnol 34, 443–456 (2007). https://doi.org/10.1007/s10295-007-0216-6

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