Elucidating Growth Coupling of Metabolites in E. coli and Characterizing Modularity.

This project seeks to increase the efficiency of rapid engineering strain design by elucidating the functionality of a modular cell. The modular cell theory insinuates that there exists a baseline cellular framework from which cellular products can be produced by inserting a designed production module into the cell. In this study, recombinant gene cloning techniques were used to design metabolite production modules that focused on increasing the production, titer, and yield of ethanol and lactate within Escherichia coli (E. coli). An auxotrophic chassis cell was created by ModCell2 and multi-objective strain design to determine the genes necessary for deletion. The resulting chassis cell and production modules were combined in a “plug in and out” fashion and then characterized for modularity. The characterization of the modular cell focused on observing a growth coupling phenomenon, characteristic of a modular cell. The growth of the cell was quantified via optical density measurements in a spectrophotometer, and the titer of the desired metabolites were measured using high performance liquid chromatography (HPLC). As a result of this study, it was determined that the modular cells for both ethanol (pHS0238) and lactate (pHS0143) exhibited growth coupling and demonstrated modular potential.

.