Purification and Characterization of a Putative Adenylosuccinate Lyase (Toy F) Involved In Toyocamycin Production

The observed increase in bacteria resistant to a variety of antibiotics is starting to grow at an alarming rate and it poses a threat to humankind. This has led to a renewed interest in a variety of fields to discover and develop molecules with novel therapeutic mechanisms in order to combat these newly emerging strains of bacteria. Two molecules that show promise as novel antibiotics are Toyocamycin and Sangivamycin, which are produced by the bacteria Streptomyces rimosus. However, they are difficult molecules to synthesize in large quantities using standard synthetic approaches. An alternative approach to production of these molecules is using the enzymes from the organism. This chemoenzymatic approach has the main challenge of identifying the enzymes involved in the natural biosynthetic pathway of the metabolites. Early work by McCarty and Bandarian has identified a cluster of genes that encode putative enzymes proposed to carry out this synthesis in S. rimosus. The work presented in this poster focuses on the purification and functional characterization of the enzyme ToyF, which is a putative adenylosuccinate lyase. ToyF was shown to convert the substrate analog adenylosuccinate into AMP, which is consistent with its predicted function.