Development of a Antibacterial Disc Diffusion Assay to Differentiate Fusarin C from Non-Fusarin C activity in Early Screening of Mutant Fusarium graminearum Extracts Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/6682x5479

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  • The discovery of novel compounds with antibacterial properties continues to be critically important.3 One potential source of such compounds is the cryptic genome of fungi known to produce biologically active molecules. A kmt6 mutant of the cereal pathogen Fusarium graminearum was previously developed through a histone H3 lysine 27 methyltransferase gene knockout, and has been shown to express hundreds of additional genes compared to wild type under the same growth conditions.1 Both the kmt6 mutant and the wild type F. graminearum produce the biologically active metabolite fusarin C, shown in Figure 1, and its analogs. Activity due to fusarin C can mask any activity of non-fusarin C compounds during early screening of extracts. Using a broad panel of bacteria, two samples were tested for differentiating activity profiles. These included an ethyl acetate extract of kmt6 growth media (i.e., the “160001E” fraction), and a more purified flash column fraction containing fusarin C and its analogs (i.e., the “fusarin C isolate”). Intriguingly, both Bacillus cereus and Staphylococcus aureus were inhibited by the 160001E extract, but were not inhibited by the fusarin C isolate. This indicates that activity from the 160001E fraction is not due to fusarin C or its analogs, but rather due to non-fusarin C compounds. With these results, a panel of 5 bacteria, including both gram negative and gram positive strains, has been developed as an early screening tool to differentiate activity between fusarin C isolates and non-fusarin C compounds from kmt6 mutant F. graminearum. This will aid in the bioassay-guided fractionation of extracts exhibiting antibacterial activity due to non-fusarin C compounds and toward the important aim of identifying novel compounds with antibacterial activity.
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