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University of Ljubljana, Biotechnical Faculty Research projects (co)funded by the Slovenian Research Agency Project Member of University of Ljubljana University of Ljubljana, Biotechnical Faculty Code J4-7162 Project Applications of aegerolysin-like proteins for detection and eradication of pests Period 1.1.2016 - 31.12.2018 Range in 2016 1,45 FTE Head Kristina Sepčić Research activity Biotechnical sciences Research Organisation Partners Agricultural institute of Slovenia Jožef Stefan Institute National Institute of Chemistry University of Ljubljana, Faculty of Medicine Abstract To meet increasing food demands and to comply with stricter environmental demands, agriculture must increase food production and quality while decreasing its detrimental ecological impact. These strategies can also be directed towards efficient control of pests, by replacing the use of first-generation chemical pesticides with novel, safer, and environmentally friendly biopesticides. Bacterial and fungal proteins belonging to the aegerolysin family have been recently found to interact with ceramide phosphoethanolamine, a membrane sphingolipid that is specific to invertebrates. Moreover, the genomes of some aegerolysin-producing fungi harbour nucleotide sequences that encode proteins with a membrane-attack-complex/ perforin (MACPF) domain. When combined, aegerolysins and these MACPF-proteins can perforate artificial and biological membranes that contain the specific lipid receptor, by forming transmembrane pore complexes. Similarly, the genomes of some aegerolysin-producing bacteria include nucleotide sequences that encode larger protein partner(s) that, when combined with aegerolysins, have membrane-perforating activity. Some of these bacterial aegerolysin-based heteromeric complexes have already been reported to have selective insecticidal properties, and have been successfully introduced into plants via genetic transformation. In the proposed project, we aim to further evaluate the potential of a battery of recombinant aegerolysins from selected bacteria and fungi, alone and in combination with their MACPF-protein partners, to act as potential bio-pesticides against some of the currently most damaging insects, and other selected invertebrates. Aegerolysin interactions with membrane lipids and soluble lipid derivatives will be further investigated, and the crystal structures of these complexes will be solved, to provide the new structural information that is necessary for further protein engineering and improvements of the pesticidal properties. The data obtained will also shed light on the currently still cryptic biological roles of these widely distributed and highly expressed proteins, the aegerolysins. Researchers link on sicris The phases of the project and their realization We are currenlly preparing and characterizing several recombinant fungal and bacterial aegerolysin proteins, and studying the interactions of these proteins with membrane lipid systems. Experiments aimed to elucidate the biological role of aegerolysins in the fungus Aspergillus niger are also in progress. We are also rearing and collecting different target pest organisms on which the toxic potential of aegerolysins will be tested. Citations for bibliographic records link on sicris