Combinatorial metabolic engineering of Escherichia coli for de novo production of 2′-fucosyllactose

Abstract

2′-Fucosyllactose (2′-FL) is a kind of fucosylated lactose of human milk oligosaccharides (HMOs). In this work, Escherichia coli MG1655 was metabolically engineered to increase 2′-FL production. The 2′-FL titer was raised from 0.0118 to 0.8062 g/L by increasing three gene copies of α1,2-fucosyltransferase (FutC) and by deleting the lon, wcaJ, lacZ, and lacI genes in the competitive pathways. Additionally, the 2′-FL titer was raised to 2.9 g/L by fusing a TrxA tag at the N-terminus of FutC, and then to 3.4 g/L by deleting glutathione reductase (Gor). Finally, the 2′-FL reached 3.3 g/L in the final strain E. coli MG27 through the de novo pathway in shake flask, and reached 10.3 g/L in a 3-L fermentor.