Combinatorial Engineering of Saccharomyces Cerevisiae for Terminal Alkene Production

Biological production of terminal alkenes has garnered a significant interest due to their industrial applications such as lubricants, detergents and fuels. Here, we engineered the yeast Saccharomyces cerevisiae to produce terminal alkenes via a one-step fatty acid decarboxylation pathway and improved the alkene production using combinatorial engineering strategies. In brief, first, we screened and characterized eight fatty acid decarboxylases (OleT) to enable and enhance alkene production in S. cerevisiae. We then developed a fatty acid-overproducing strain to boost the precursor availability, which could enhance the metabolic flux and resulted in a higher production titer. We then improved the enzyme cofactor accumulation through cofactor genetic engineering. We then enhanced the cell growth in rich medium and tuned the enzyme expression by optimizing the combinations of the promoters and plasmids. Finally, we further increased the alkene production by optimizing the culturing conditions in bioreactors. This study represents the first report of terminal alkene biosynthesis in the yeast S. cerevisiae, and we envision that the abovementioned combinatorial engineering approaches could provide insights into devising engineering strategies to improve the production of fatty acid-derived biochemicals in S. cerevisiae.