Production of borneol, camphor, and bornyl acetate using engineered Saccharomyces cerevisiae

Microbial production of bicyclic monoterpenes is of great interest because their production primarily utilizes non-sustainable resources. Here, we report an engineered <i>Saccharomyces cerevisiae</i> yeast that produces bicyclic monoterpenes, including borneol, camphor, and bornyl acetate. The engineered yeast expresses a bornyl pyrophosphatase synthase from <i>Salvia officinalis</i> fused with mutated farnesyl pyrophosphate synthase from <i>S</i>. <i>cerevisiae</i> and two mevalonate pathway enzymes (an acetoacetyl-CoA thiolase/hydroxymethylglutaryl-CoA [HMG-CoA] reductase and an HMG-CoA synthase) from <i>Enterococcus faecalis</i>. The yeast produced up to 23.0 mg/L of borneol in shake-flask fermentation. By additionally expressing borneol dehydrogenase from <i>Pseudomonas</i> sp. TCU-HL1 or bornyl acetyltransferase from <i>Wurfbainia villosa</i>, the engineered yeast produced 23.5 mg/L of camphor and 21.1 mg/L of bornyl acetate, respectively. This is the first report of heterologous production of camphor and bornyl acetate.