PROJECT Summary

​​​

Adaptive Laboratory Evolution (ALE) was employed as a powerful tool to shape microorganisms for industrial applications. This project focused on using ALE to optimize Escherichia coli for growth and for the utilization of sucrose as a carbon source. For these purposes, E. coli strains were first engineered with the csc genes (cscB, cscK, cscA) to enable sucrose uptake. The genetically modified strains were then subjected to an ALE regimen, during which they were propagated to gradually consume increasing concentrations of sucrose in synthetic medium. Mutations that accumulated during the evolutionary process were identified through next-generation genome sequencing and, using the EasyGuide CRISPR platform, were subsequently reverse engineered into the parental strains. Finally, the strains that had been optimized for anaerobic growth on sucrose were subjected to metabolic engineering for polyhydroxyalkanoate (PHA) synthesis, yielding candidate strains ideally suited for bioplastic production from sugarcane feedstock.