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A novel actinomycete strain de-replication approach based on the diversity of polyketide synthase an.pdf

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A novel actinomycete strain de-replication approach based on the diversity of polyketide synthase an

Appl Microbiol Biotechnol (2005) 67: 795–806 DOI 10.1007/s00253-004-1828-7 APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY Angel Ayuso . Desmond Clark . Ignacio González . Oscar Salazar . Annaliesa Anderson . Olga Genilloud A novel actinomycete strain de-replication approach based on the diversity of polyketide synthase and nonribosomal peptide synthetase biosynthetic pathways Received: 13 July 2004 / Revised: 25 October 2004 / Accepted: 30 October 2004 / Published online: 13 January 2005 # Springer-Verlag 2005 Abstract The actinomycetes traditionally represent one of the most important sources for the discovery of new metabolites with biological activity; and many of these are described as being produced by polyketide synthases (PKS) and nonribosomal peptide synthetases (NRPS). We present a strain characterization system based on the metabolic potential of microbial strains by targeting these biosynthetic genes. After an initial evaluation of the existing bias derived from the PCR detection in a well defined biosynthetic sys- tems, we developed a new fingerprinting approach based on the restriction analysis of these PKS and NRPS amplified sequences. This method was applied to study the distribu- tion of PKS and NRPS biosynthetic systems in a collection of wild-type actinomycetes isolated from tropical soil sam- ples that were evaluated for the production of antimicrobial activities. We discuss the application of this tool as an al- ternative characterization approach for actinomycetes and we comment on the relationship observed between the presence of PKS-I, PKS-II and NRPS sequences and the antimicrobial activities observed in some of the microbial groups tested. Introduction In recent decades, natural-product screening programs de- voted an enormous effort to discovering new microbial secondary metabolites with interesting biological activities. One of the most important steps in the drug discovery pro- cess from microbial natural products is the selection of new mi