De novo biosynthesis of 2ʹ‐fucosyllactose by bioengineered Corynebacterium glutamicum

Abstract 2ʹ‐Fucosyllactose (2ʹ‐FL) which is well‐known human milk oligosaccharide was biotechnologically synthesized using engineered Corynebacterium glutamicum , a GRAS microbial workhorse. By construction of the complete de novo pathway for GDP‐L‐fucose supply and heterologous expression of Escherichia coli lactose permease and Helicobacter pylori α‐1,2‐fucosyltransferase, bioengineered C. glutamicum BCGW_TL successfully biosynthesized 0.25 g L −1 2ʹ‐FL from glucose. The additional genetic perturbations including the expression of a putative 2ʹ‐FL exporter and disruption of the chromosomal pfkA gene allowed C. glutamicum BCGW_cTTLEΔP to produce 2.5 g L −1 2ʹ‐FL batchwise. Finally, optimized fed‐batch cultivation of the BCGW_cTTLEΔP using glucose, fructose, and lactose resulted in 21.5 g L −1 2ʹ‐FL production with a productivity of 0.12 g L −1 •h, which were more than 3.3 times higher value relative to the batch culture of the BCGW_TL. Conclusively, it would be a groundwork to adopt C. glutamicum for biotechnological production of other food additives including human milk oligosaccharides.