Construction and application of 3-fucosyllactose whole-cell biosensor for high-throughput screening of overproducers

[Display omitted] •A growth-coupled 3-fucosyllactose biosensor based on α-L-fucosidase is developed.•Inactivated ManX removing growth interference of mannose could optimize biosensor.•3-fucosyllactose hyperproducers and new high-producing mechanisms are obtained.•The strategy provides a new idea for constructing other HMOs biosensors. Biosensor-based high-throughput screening is efficient for improving industrial microorganisms. There is a severe shortage of human milk oligosaccharides (HMOs) biosensors. This study established a 3-fucosyllactose (3-FL, a kind of HMOs) whole-cell biosensor by coupling cell growth with production. To construct and optimize the biosensor, an Escherichia coli 3-FL producer was engineered by deleting the manA, yihS and manX genes, directing the mannose flux solely to 3-FL synthesis. Then, an α-L-fucosidase was introduced to hydrolyze 3-FL to fucose which was used as the only carbon source for cell growth. Using the biosensor, the 3-FL production of a screened mutant was improved by 25 % to 42.05 ± 1.28 g/L. The productivity reached 1.17 g/L/h, the highest level reported by now. The csrB mutant obtained should be a new clue for the 3-FL overproduction mechanism. In summary, this study provided a novel approach to construct HMOs biosensors for strain improvement.