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Crystal Structure of alpha1,3-FucosyltransferaseCrystal Structure of alpha1,3-Fucosyltransferase
Structural highlights
Function[FUCT_HELPX] Involved in the biosynthesis of the Lewis X (LeX) trisaccharide of the lipopolysaccharide (LPS) O-antigen. Catalyzes the addition of fucose in alpha 1-3 linkage to Gal-beta-1-4-GlcNAc-beta-O-R (LacNAc-R) type II acceptor.[1] [2] [3] [4] [5] Publication Abstract from PubMedFucosylated 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 (>10(7) 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 alpha1,3-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 the M32 mutant revealed that the S45F mutation generates a clamp-like structure that appears to improve binding of the galactopyranose ring of the acceptor substrate. Moreover, molecular dynamic simulations reveal that helix alpha5, is more mobile in the M32 mutant, possibly explaining its high fucosylation activity. Directed evolution of an alpha1,3-fucosyltransferase using a single-cell ultrahigh-throughput screening method.,Tan Y, Zhang Y, Han Y, Liu H, Chen H, Ma F, Withers SG, Feng Y, Yang G Sci Adv. 2019 Oct 9;5(10):eaaw8451. doi: 10.1126/sciadv.aaw8451. eCollection 2019, Oct. PMID:31633018[6] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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