Discovery of New\nCyclic Lipodepsipeptide Orfamide\nN via Partnership with Middle School Students from the Boys and Girls\nClub

We established a university-community partnership with\nthe Boys\nand Girls Clubs of Chicago (BGCC)named <i>Chicago Antibiotic\nDiscovery La</i>bto involve middle school students in\nantibiotic discovery research. In the course of working with a cohort\nof students from the BGCC, one student isolated a Pseudomonas\nidahonensis bacterium from a goose feces sample that\nproduced a new cyclic lipodepsipeptide, which was characterized as\norfamide N. Orfamide N is composed of ten mixed D/L-amino acids and\na (<i>Z</i>)-3<i>R</i>-hydroxyhexadec-9-enoic\nacid residue. The planar structure of orfamide N was elucidated by\none-dimensional (1D) and two-dimensional nuclear magnetic resonance\n(2D NMR), electrospray ionization-mass spectrometry (ESI-MS), and\nozone-induced dissociation mass spectrometry (OzID-MS). The absolute\nconfiguration was determined by advanced Marfey’s analysis,\nphylogenetic analysis of C-domains within the orfamide N biosynthetic\ngene cluster, and chiral high-performance liquid chromatography (HPLC)\nanalysis of the hydrolyzed and reduced lipid tail. Orfamide N was\ncytotoxic against human melanoma and human ovarian cancer cells with\nIC<sub>50</sub> values of 11.06 and 10.50 μM, respectively.\nOverall, we demonstrated it is possible to integrate educational outreach\nwith high-end natural product discovery while strengthening the relationship\nbetween the university and the community it serves.