In this study, stepwise increases in linalool production were obtained by combining metabolic engineering and process optimization of an unconventional oleaginous yeast Yarrowia lipolytica. The linalool synthetic pathway was successfully constructed by heterologously expressing a codon-optimized linalool synthase gene from Actinidia arguta in Y. lipolytica. To enhance linalool productivity, key genes involved in the mevalonate pathway were overexpressed in different combinations. Moreover, the overexpression of mutant ERG20(F88W-N119W) gene resulted in further linalool production. A maximum linalool titre of 6.96 +/- 0.29 mg/L was achieved in shake flasks, which was the highest level ever reported in yeasts. (C) 2017 Elsevier Ltd. All rights reserved.