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Antibiotic-resistance has emerged as one of the major public health problems of the 21st century. The rate at which novel antibiotics are discovered has declined for over thirty years, while simultaneously the number of antibiotic resistant strains of harmful bacteria has risen. Natural products or synthetic variations of such account for about two-thirds of all FDA approved drugs. However, the standard drug discovery platform – growing and extracting liquid bacterial cultures – has become ineffective as 99% of microbes cannot be grown in the laboratory. A novel culture-free technique allows access by heterologous expression to the biosynthetic pathways of these unculturable microbes by extracting DNA directly from the environment. In the current work, three heterologously expressed bacterial type II polyketide pathways were examined for the production of new molecules. Extraction of liquid cultures and analysis by LCMS revealed the presence of clone-specific metabolites in all three pathways. Purification by HPLC and silica flash chromatography were attempted with varying degrees of success, and the product of one incomplete pathway was successfully characterized by 1-D and 2-D NMR.
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Low-Beinart, Lila Reed, "The Isolation and Characterization of Type II Polyketides from Soil Metagenomes; Addressing the shortage of effective antibiotics by a new method of antibiotic discovery" (2014). Senior Projects Spring 2014. 318.