Identification of the Streptomyces aculeolatus proteins involved in the biosynthesis of bioactive compound by a comparative 2D electrophoresis approach

Abstract

Event Abstract Back to Event Identification of the Streptomyces aculeolatus proteins involved in the biosynthesis of bioactive compound by a comparative 2D electrophoresis approach Ana F. Fernandes1, 2, 3*, Lígia Costa1, 2, 3*, Ilda Santos-Sanches1, Susana P. Gaudêncio1, 3 and M. Gabriela Almeida2, 4 1 UCIBIO-REQUIMTE, Department of Life Sciences, Faculty of Science and Technology, Universidade NOVA de Lisboa, Portugal 2 UCIBIO-REQUIMTE, Department of Chemistry, Faculty of Science and Technology, Universidade NOVA de Lisboa, Portugal 3 LAQV-REQUIMTE, Department of Chemistry, Faculty of Science and Technology, Universidade NOVA de Lisboa, Portugal 4 Centro de Investigação Interdisciplinar Egas Moniz - Instituto Superior de Ciências de Saúde Egas Moniz, Portugal Actinomycetes Streptomyces aculeolatus belong to MAR4 streptomycete lineage and are known to produce hybrid isoprenoids including a naphthoquionone moiety, such as napyradiomycin and marinone families, a phthalazinone core as in azamerone class, and a phenazine moiety as in lavanducyanin and marinophenazine families.[1] These compounds have great bioactive potential as antibacterial, antifungal, and anticancer agents making them attractive leads to the pharmaceutical industry.[1],[2],[3] A total of 421 actinomycetes were isolated from marine sediments collected from the Madeira Archipelago, six of these belong to the S. aculeolatus specie, as determined by a phylogenetic analysis based on the sequencing of the gene 16S rRNA. These strains (coded PTM-29, PTM-81, PTM-129, PTM-346, PTM-398 and PTM-420) revealed the production of secondary metabolites with distinct bioactivities. In this context, we aim at characterizing the proteins involved in the synthesis of the main bioactive compounds at the strains PTM-29, PTM-129, PTM-346 and PTM-398. To achieve this goal, we are following a differential proteomic analysis based in two-dimensional electrophoresis (2DE) followed by mass spectrometry techniques (MS). The total and extracellular proteome of S. aculeolatus strains were analysed in the pH range 4-7, in 12,5% acrylamide gels stained with Colloidal Commassie Blue. Preliminary results showed similar protein profiles for the total proteomes, with most of the spots in the pH range 5-7 and a high molecular weight (above 50 kDa). Curiously, the extracellular proteomes have shown to be very different among strains, also presenting high molecular weight proteins. As future work, we intend to analyse in detail the 2D gels from the S. aculeolatus strains in study and identify the differentially expressed proteins by mass spectrometry (MS) techniques. Acknowledgements This research is the result of financial support from Fundação para a Ciência e a Tecnologia (FCT) and FEDER through grant PTDC/QUI-QUI/119116/2010, IF/00700/2014, PEst-C/EQB/LA0006/2013, UID/QUI/50006/2013 and UID/Multi/04378/2013, and the European Union 7th Framework Programme (FP7/2007-2013) under grant agreement PCOFUND-GA-2009-246542 and 269138-NanoGuard, and grant Instruct Access1153.