Chemical investigation, genomic mining and effects on Langmuir membrane models of cytotoxic metabolites produced by Penicillium limosum CMLD 19

Abstract

The investigation of metabolites produced by microorganisms from Penicillium genus, especially those related to species that are still little explored, represents an important pathway for the discovery of molecules with anticancer potential. Penicillium limosum represents a fungus which has been little studied from the metabolomic and genomic point of view. Also, the biological potential of its metabolites seem to be underexplored. Preliminary studies carried out with the endophytic strain P. limosum CMLD 19 indicated significant cytotoxic activity from its metabolic extracts when they were tested against human leukemia cell lines. Moreover, mining the genome of CMLD 19, it was found a biosynthetic gene cluster (BGC) with 55 % of similarity to a BCG that encodes the production of enzymes responsible for the biosynthesis of structural analogues of brefeldin A. This polyketide macrolide is a natural product that impacts on the growth of mammalian cells and has been widely investigated for the development of chemotherapy drugs against neoplasms, such as leukemia. After metabolomic analysis based on mass spectrometry data, there was indeed the detection of brefeldin A and its co-produced congeners in organic extracts of CMLD 19. Therefore, the main goal of this project is to reach comprehensive understandings about the chemical diversity produced by this microorganism owing to encounter molecular targets with anti-leukemic potential, such as unknown congeners of brefeldin A. In order to do so, the project strategy involves the integration of modern metabolomic analyses, combined with the genome mining of the microorganism, as well as to perform cytotoxicity tests of its metabolites against leukemia cell lines. The screening for bioactive molecules and the comprehension of the mechanism of action of these molecular targets will be also investigated using membrane models mimicked by Langmuir films. The combination of these different approaches implemented for the prospection of P.limosum metabolism could uncover new bioactive natural products that may contribute to the struggle against leukemias.