Extraction and Partial Purification of an Antibiotic-like Compound From the Soil Bacterium Rhoddococcus Strain KCHXC3

Rhodococcus bacteria have many secondary metabolic pathways that may produce novel natural products. The bacterium Rhodococcus strain KCHXC3 was isolated from a soil sample collected near Kingsport, Tennessee and was found to produce an inhibitory compound active against a broad array of bacterial species, including the Gram negative pathogen Shigella . The aim of this research is to extract and purify the compound for future structure elucidation. A mixture of compounds from 3 month old agar plates inoculated with strain KCHXC3 was extracted using ethyl acetate. The crude extract was then partially purified utilizing a Sephadex LH-20 column, followed by an analytical NH ₂ HPLC column. This purification resulted in a dried crystalline-like active compound that is white in color and needle-like in shape. Structural studies such as NMR and GC-MS were performed in order to determine the purity of this solid.