Microbial-produced polyketides are promising biosustainable molecules with uses ranging from high-value pharmaceuticals to biorenewable chemical precursors. <i>S. cerevisiae</i> is particularly suitable for polyketide production due to a high tolerance to otherwise toxic products, in addition to a GRAS rating by the FDA. To maximize polyketide production, previous work in our lab primarily considered single intervention strategies at a time. Our recent efforts to combine these strategies have uncovered unexpected complexity in predicting outcomes. By coupling computational modeling with CRISPR-Cas9, we can identify and subsequently modify promising genes rapidly and efficiently, and compare many strains with multiple modifications simultaneously. With these methods, we are evaluating combinations of gene knockouts and knockins that influence disparate regions of cellular regulation and metabolism. Our work considers central carbon metabolism (to increase key polyketide precursors such as acetyl- and malonyl-CoA), maximization of theoretical carbon yield, transport processes, and global yeast transcriptional networks. Tailoring other factors such as media and growth conditions, promoter selection, enzyme engineering, and enzyme copy number has also increased polyketide titer and yield. Efficacy of these strategies are tested via production of tracetic acid lactone (TAL), a simple polyketide requiring only one heterologous enzyme (2-PS). TAL itself is a useful molecule for conversion to sorbic acid and 1,3-butanediol.

Combinatorial strategies for optimizing polyketide production in <i>Saccharomyces cerevisiae</i>

253rd National Meeting (2017)

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Combinatorial strategies for optimizing polyketide production in Saccharomyces cerevisiae