High-throughput method for screening alpha-1,2-fucosyltransferase and screened mutant

A high-throughput method for screening α-1,2-fucosyltransferase, comprising the steps of:

• Step S1:
• constructing an indicator strain of 2′-fucosyllactose, wherein the indicator strain comprises a first expression vector and a second expression vector, the first expression vector comprises an α-L-fucosidase coding gene, the second expression vector comprises a lac operon and a fluorescent reporter gene, and α-L-fucosidase degrades 2′-fucosyllactose into lactose, and regulates, together with the lac operon, the expression of the fluorescent reporter gene, so that the content of 2′-fucosyllactose is correlated with the fluorescence intensity;
• Step S2:
• constructing a chassis strain, wherein the chassis strain expresses an L-fucokinase coding gene and an α-1,2-fucosyltransferase mutant library;
• Step S3:
• embedding the chassis strain in single-cell droplets and culturing the droplets, to obtain a droplet microfluidic chip, and culturing the droplets in a medium containing fucose and lactose;
• and
• Step S4:
• subjecting the cell suspension of the indicator strain to one-to-one fusion with the droplets, co-culturing, microfluidically sorting the droplets according to the fluorescence intensity in the fused droplets, to obtain a chassis strain comprising a forward mutant of α-1,2-fucosyltransferase.