Directed evolution of an α1,3-fucosyltransferase using a single-cell ultrahigh-throughput screening method.

ABSTRACT: Fucosylated glycoconjugates are involved in a variety of physiological and pathological processes. However, economical production of fucosylated drugs and prebiotic supplements has been hampered by the poor catalytic efficiency of fucosyltransferases. Here, we developed a fluorescence-activated cell sorting system that enables the ultrahigh-throughput screening (>10⁷ mutants/hour) of such enzymes and designed a companion strategy to assess the screening performance of the system. After three rounds of directed evolution, a mutant M32 of the α1,3-FucT from Helicobacter pylori was identified with 6- and 14-fold increases in catalytic efficiency (k _cat/K _m) for the synthesis of Lewis x and 3'-fucosyllactose, respectively. The structure of

SUBMITTER: Tan Y 

PROVIDER: S-EPMC6785251 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature