Discovery of New Cyclic Lipodepsipeptide Orfamide N via Partnership with Middle School Students from the Boys and Girls Club

We established a university-community partnership with the Boys and Girls Clubs of Chicago (BGCC)-named Chicago Antibiotic Discovery Lab-to involve middle school students in antibiotic discovery research. In the course of working with a cohort of students from the BGCC, one student isolated a Pseudomonas idahonensis bacterium from a goose feces sample that produced a new cyclic lipodepsipeptide, which was characterized as orfamide N. Orfamide N is composed of ten mixed D/L-amino acids and a (Z)-3R-hydroxyhexadec-9-enoic acid residue. The planar structure of orfamide N was elucidated by one-dimensional (1D) and two-dimensional nuclear magnetic resonance (2D NMR), electrospray ionization-mass spectrometry (ESI-MS), and ozone-induced dissociation mass spectrometry (OzID-MS). The absolute configuration was determined by advanced Marfey's analysis, phylogenetic analysis of C-domains within the orfamide N biosynthetic gene cluster, and chiral high-performance liquid chromatography (HPLC) analysis of the hydrolyzed and reduced lipid tail. Orfamide N was cytotoxic against human melanoma and human ovarian cancer cells with IC50 values of 11.06 and 10.50 μM, respectively. Overall, we demonstrated it is possible to integrate educational outreach with high-end natural product discovery while strengthening the relationship between the university and the community it serves.