Bacteria are developing resistance to antibiotics at a faster rate than the successful discovery and production of antibiotics, challenging researchers with the task of developing efficient antibiotics that have minimal risk of resistance development. Soil, which contains a number of bacteria, serves as an excellent source to sample species for antagonistic compounds. In this study, a strain of Pseudomonas putida, isolated from soil near Heidelberg University, screened positive for the production of antibiotic molecules against ESKAPE pathogens. The primary antibiotic compound(s), including pyoverdine and an unknown lipid, are both side and direct products of the Ferric-Pyoverdine biosynthetic pathway (Ye et al, 2014). This pathway is highly dependent on the presence, or lack thereof, of organic nitrates and free Fe<sup>3+</sup> (Ringel and Bruser, 2018). Using ethylenediaminetetraacetic acid (EDTA) and various iron salts, the optimal concentration of iron necessary for the production of pyoverdines was established. UV-Vis spectroscopy and fluorescent assaying were used to confirm the presence of pyoverdines. All lipids present following pyoverdine synthesis were extracted and separated using a series of organic and aqueous phase extractions. Primary analysis of these compounds using HPLC, IR spectroscopy, and affinity chromatography were used to separate the lipids based on polarity and carbon chain length. Each separation was tested for antagonistic activity using a disk-diffusion assay, allowing for the antagonistic compounds to be identified. Results show that the antibiotic activity of Pseudomonas putida is a combinatory result of both the pyoverdine molecule itself and a range of lipid products.

Extraction and characterization of antagonistic compounds from the Ferric-Pyoverdine biosynthetic pathway in pseudomonas putida soil strains

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.