5ZOI

Crystal Structure of alpha1,3-Fucosyltransferase

Summary for 5ZOI

• Entry DOI: 10.2210/pdb5zoi/pdb
• Descriptor: Alpha-(1,3)-fucosyltransferase FucT, [[(2S,3R,4S,5R)-5-(2-azanyl-6-oxidanylidene-1H-purin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl] [(2S,3S,4S,5R,6R)-6-methyl-3,4,5-tris(oxidanyl)oxan-2-yl] hydrogen phosphate (2 entities in total)
• Functional Keywords: enzyme, transferase
• Biological source: Helicobacter pylori (Campylobacter pylori)
• Total number of polymer chains: 2
• Total formula weight: 99588.30

Authors

Tan, Y.,Yang, G. (deposition date: 2018-04-13, release date: 2019-06-26, Last modification date: 2024-03-27)

Primary citation

Tan, Y.,Zhang, Y.,Han, Y.,Liu, H.,Chen, H.,Ma, F.,Withers, S.G.,Feng, Y.,Yang, G.
Directed evolution of an alpha 1,3-fucosyltransferase using a single-cell ultrahigh-throughput screening method.
Sci Adv, 5:eaaw8451-eaaw8451, 2019

PubMed 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 (>10 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 was identified with 6- and 14-fold increases in catalytic efficiency ( / ) 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 α5, is more mobile in the M32 mutant, possibly explaining its high fucosylation activity.

PubMed: 31633018
DOI: 10.1126/sciadv.aaw8451

Experimental method

X-RAY DIFFRACTION (3.19 Å)