Designing and Implementing an Assay for the Detection of Rare and Divergent NRPS and PKS Clones in European, Antarctic and Cuban Soils

Varování

Publikace nespadá pod Lékařskou fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.

Název česky	Návrh a využití metody detekce nezvyklých a odlišných NRPS a PKS klonů v evropských, antarktických a kubánských půdách
Autoři	AMOS G.C.A. BORSETTO C. LASKARIS P. KRSEK Martin BERRY A.E. NEWSHAM K.K. CALVO-BADO L. PEARCE D.A. VALLIN C. WELLINGTON E.M.H.
Rok publikování	2015
Druh	Článek v odborném periodiku
Časopis / Zdroj	Plos one
Fakulta / Pracoviště MU	Přírodovědecká fakulta
Citace
www	http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0138327
Doi	http://dx.doi.org/10.1371/journal.pone.0138327
Obor	Mikrobiologie, virologie
Klíčová slova	antibiotics; metagenomics; NRPS; PKS; actinobacteria; proteobacteria
Popis	The ever increasing microbial resistome means there is an urgent need for new antibiotics. Metagenomics is an underexploited tool in the field of drug discovery. In this study we aimed to produce a new updated assay for the discovery of biosynthetic gene clusters encoding bioactive secondary metabolites. PCR assays targeting the polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS) were developed. A range of European soils were tested for their biosynthetic potential using clone libraries developed from metagenomic DNA. Results revealed a surprising number of NRPS and PKS clones with similarity to rare Actinomycetes. Many of the clones tested were phylogenetically divergent suggesting they were fragments from novel NRPS and PKS gene clusters. Soils did not appear to cluster by location but did represent NRPS and PKS clones of diverse taxonomic origin. Fosmid libraries were constructed from Cuban and Antarctic soil samples; 17 fosmids were positive for NRPS domains suggesting a hit rate of less than 1 in 10 genomes. NRPS hits had low similarities to both rare Actinobacteria and Proteobacteria; they also clustered with known antibiotic producers suggesting they may encode for pathways producing novel bioactive compounds. In conclusion we designed an assay capable of detecting divergent NRPS and PKS gene clusters from the rare biosphere; when tested on soil samples results suggest the majority of NRPS and PKS pathways and hence bioactive metabolites are yet to be discovered.