Investigating antimicrobial mechanisms-of-action using automated genome-wide and gene-specific editing

Some heterologous metabolic pathway intermediates can inhibit microbial growth and stifle production capabilities of bioindustrial strains through poorly understood mechanisms. We have developed a novel approach to investigating the mechanisms of action of small inhibitory molecules using automated, trackable genome engineering. A combination of genome-wide and gene-specific edits allowed us to identify genes and individual residues implicated in biological responses to p-coumaric acid and resveratrol and provided target leads for engineering tolerance or enhanced sensitivity to these compounds.