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734840 
Technical Report 
Engineer novel anticancer bioagents 
Wang, S; Wesener, S; Portharla, V; Cheng, Y 
2009 
University of Wisconsin at Milwaukee 
Milwaukee, Wisconsin 
Our research in the first contract-year has resulted in: (1) the confirmation of a redox enzyme (encoded by depH) responsible for a critical disulfide bond formation as the final step in FK228 biosynthesis (a publication is appended), (2) the identification of an unexpected pathway regulatory gene depR (previously annotated as orf18) (a manuscript is in preparation), and (3) the successful reconstitution of FK228 biosynthetic gene cluster on three vectors in an E. coli strain, and the detection of heterologous FK228 production in E. coli under both aerobic and anaerobic growth conditions (another manuscript is in preparation as well). Attempt to integrate the complete FK228 biosynthetic gene cluster into the E. coli chromosome for stable functioning without antibiotic selection is in progress. We therefore have achieved the project milestone with some adjustments of the experimental approach and research content. 
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