Metabolic engineering of Escherichia coli BL21(DE3) cocultured with glucose and xylose for efficient production of 2'-fucosyllactose.

2'-Fucosyllactose (2'-FL) is the most abundant human milk oligosaccharide (HMO) and has been approved to be commercially added to infant formula. Microbial synthesis from exogenous lactose via metabolic engineering is currently the major approach to production of 2'-FL. Replacement of lactose with cheaper sugars such as glucose and sucrose has been studied to reduce the production costs. Herein, Escherichia coli BL21(DE3) was engineered to produce 2'-FL by co-culture with glucose and xylose, the main components of lignocellulosic biomass. Firstly, synthetic pathway of lactose from xylose and glucose was constructed by introducing a lactose-forming enzyme, strengthening xylose uptake pathway, and weakening glucose metabolic pathway. Then, a highly-active α1,2-fucosyltransferase BKHT was introduced to produce 2'-FL and GDP-fucose supply was enhanced to increase 2'-FL production. As a result, when cocultured with glucose and xylose, the engineered strain produced 6.53 g/L and 27.53 g/L of 2'-FL by shake-flask and fed-batch cultivation, respectively.