High production of fatty alcohols in Yarrowia lipolytica by coordination with glycolysis

Abstract

Fatty alcohol biosynthesis by oleaginous microbes was a promising alternative to the petroleum or other non-renewable resources-based process. However, low titer and yield hamper the further industrial and commercial applications. Here, we developed an efficient strategy to coordinate fatty alcohol with glycolysis which achieved a ‘pull-and-push’ effect to improve fatty alcohol production. Transcript profiling indicated that genes in carbohydrate metabolism were up-regulated significantly in response to high fatty alcohol production. Based on it, 11 glycolysis promoters were screening to express fatty acyl-CoA reductase (FAR) to relate the fatty alcohol production with the up-regulated carbohydrate metabolism, and the fatty alcohol production reached 557 mg/L when FAR was expressed by the promoter of PFBAin. RNA-seq and qRT-PCR analysis demonstrated that a ‘pull-and-push’ effect caused by the coordination system dynamically enhanced the product synthesis flux from top to bottom, which was also testified and intensified by doubled glucose concentration. After manipulating structural and regulatory genes of lipid metabolism, the final strain achieved up to 5.75 g/L fatty alcohol production from modified YPD medium (containing 91 g/L glucose) in shake flasks, which represented the highest titer reported to date. This work offered a feasible and effective reference for dynamic manipulation of fatty acid-derived chemicals synthesis.