DISCOVERING THE INDUCAMIDE BIOSYNTHETIC PATHWAY

Jennifer Korchak, Amy Lane

Department of Chemistry, University of North Florida, Jacksonville FL, USA

Fu et al. reported the discovery of novel anticancer inducamides A-C from a marine derived Streptomyces sp. strain SNC109-M3 that was obtained through a selective mutation on the β subunit of RNA polymerase that stimulated inducamide production. Inducamide C exhibits the strongest anticancer properties. The goal of the current project is to characterize the genes required for inducamide production by Streptomyces.  We determined the complete 9.8 million base-pair genome of SNC109-M3 in order to locate the hypothesized inducamide gene cluster. This revealed an approximately 18 kilobase-pair gene cluster encoding several proteins with hypothesized roles in inducamide biosynthesis. After construction of a clone library containing colonies of E. coli with 50 kilobase-pairs of randomly fragmented SNC109-M3 DNA ligated into a cosmid, PCR screening will be performed to locate the library member that contains the putative biosynthetic gene cluster. The gene cluster will be expressed in a heterologous Streptomyces host. After integration into the Streptomyces genome, production of inducamides will be quantified through LC/MS and HPLC-UV. Through the execution of the aforementioned methods, the establishment of the biosynthetic pathway for inducamides A-C will set the stage for generation of inducamide analogs, with the future goal being to synthesize increased efficiency anticancer compounds.