In vitro enzyme analysis guides production of a natural product analog

Polyketides, a structurally diverse class of natural products, are a rich source of highly effective pharmaceuticals. As such, the development of methods to produce polyketide analogs with potentially improved pharmacokinetic or toxicological properties is a research area of great interest. We will describe the in vitro analysis of a lipomycin synthase subunit, revealing the unexpected broad substrate specificity of the enzyme. We will further discuss how modifications to the culture medium can be used to modulate intracellular acyl-CoA concentrations in vivo, facilitating the production of a novel lipomycin analog by the native producer, Streptomyces auereofaciens Tü117 (2). These results highlight the utility of using in vitro enzyme analysis to determine intrinsic substrate specificity and thereby inform subsequent efforts to produce natural product analogs.