Alpha-Terpineol production from an engineered Saccharomyces cerevisiae cell factory

Alpha-Terpineol ([alpha]-Terpineol), a C.sub.10 monoterpenoid alcohol, is widely used in the cosmetic and pharmaceutical industries. Construction Saccharomyces cerevisiae cell factories for producing monoterpenes offers a promising means to substitute chemical synthesis or phytoextraction. [alpha]-Terpineol was produced by expressing the truncated [alpha]-Terpineol synthase (tVvTS) from Vitis vinifera in S. cerevisiae. The [alpha]-Terpineol titer was increased to 0.83 mg/L with overexpression of the rate-limiting genes tHMG1, IDI1 and ERG20.sup.F96W-N127W. A GSGSGSGSGS linker was applied to fuse ERG20.sup.F96W-N127W with tVvTS, and expressing the fusion protein increased the [alpha]-Terpineol production by 2.87-fold to 2.39 mg/L when compared with the parental strain. In addition, we found that farnesyl diphosphate (FPP) accumulation by down-regulation of ERG9 expression and deletion of LPP1 and DPP1 did not improve [alpha]-Terpineol production. Therefore, ERG9 was overexpressed and the [alpha]-Terpineol titer was further increased to 3.32 mg/L. The best [alpha]-Terpineol producing strain LCB08 was then used for batch and fed-batch fermentation in a 5 L bioreactor, and the production of [alpha]-Terpineol was ultimately improved to 21.88 mg/L. An efficient [alpha]-Terpineol production cell factory was constructed by engineering the S. cerevisiae mevalonate pathway, and the metabolic engineering strategies could also be applied to produce other valuable monoterpene compounds in yeast.