Abstract

2′-Fucosyllactose (2′-FL) has great application value as a nutritional component and the whole cell biosynthesis of 2′-FL has become the focus of current research. Yarrowia lipolytica has great potential in oligosaccharide synthesis and large-scale fermentation. In this study, systematic engineering of Y. lipolytica for efficient 2′-FL production was performed. By fusing different protein tags, the synthesis of 2′-FL was optimized and the ubiquitin tag was demonstrated to be the best choice to increase the 2′-FL production. By iterative integration of the related genes, increasing the precursor supply, and promoting NADPH regeneration, the 2′-FL synthesis was further improved. The final 2′-FL titer, 41.10 g/L, was obtained in the strain F5-1. Our work reports the highest 2′-FL production in Y. lipolytica, and demonstrates that Y. lipolytica is an efficient microbial chassis for the synthesis of oligosaccharides.

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