9eu4

From Proteopedia

GH29A alpha-L-fucosidase

Structural highlights

9eu4 is a 2 chain structure with sequence from Bacteroides fragilis. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:	X-ray diffraction, Resolution 2.33Å
Ligands:	,
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

Q5LAD6_BACFN Alpha-L-fucosidase is responsible for hydrolyzing the alpha-1,6-linked fucose joined to the reducing-end N-acetylglucosamine of the carbohydrate moieties of glycoproteins.[ARBA:ARBA00004071]

Publication Abstract from PubMed

GH29A alpha-l-fucosidases (EC 3.2.1.51) catalyze the release of alpha-l-fucosyl moieties from the nonreducing end of glycoconjugates by hydrolysis and some also catalyze transglycosylation. The latter is particularly interesting with regard to designing enzymatic synthesis of human milk oligosaccharides (HMOs). We combined the bioinformatics tool conserved unique peptide patterns (CUPP) and phylogenetic clustering to discover new microbial GH29A alpha-l-fucosidases of the underexplored CUPP group GH29:13.1. Three uncharacterized bacterial enzymes (EaGH29, SeGH29, and PmGH29) and two previously identified GH29A alpha-l-fucosidases (BF3242 and TfFuc1) were selected for reaction optimization, biochemical, and structural characterization. Kinetics, pH-temperature optima, and substrate preference for 2-chloro-4-nitrophenyl-alpha-l-fucopyranoside (CNP-alpha-l-Fuc) and 2'-fucosyllactose (2'FL) were determined. Transglycosylation was favored at high neutral to alkaline pH, especially for EaGH29, SeGH29, TfFuc1, and BF3242, mainly because hydrolysis was decreased. The alpha-l-fucosidases exhibited medium regioselectivity in transglycosylation, generally forming two out of five detected lacto-N-fucopentaose (LNFP) isomers from 2'FL and lacto-N-tetraose (LNT). Alkaline pH also affected the transglycosylation product regioselectivity of SeGH29, which was also affected by a Leu/Phe exchange in the acceptor binding site. New crystal structures of TfFuc1 and BF3242 showed congruence in active site topology between these two enzymes and contributed to understanding the function of GH29A alpha-l-fucosidases. Notably, the structural data provide new insight into the role of an Asn residue located between the two catalytic residues in the active site.

Structural elucidation and characterization of GH29A alpha-l-fucosidases and the effect of pH on their transglycosylation.,Yang Y, Holck J, Thorhallsson AT, Hunt CJ, Yang H, Morth JP, Meyer AS, Zeuner B FEBS J. 2025 Feb;292(3):653-680. doi: 10.1111/febs.17347. Epub 2024 Dec 10. PMID:39658312^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Yang Y, Holck J, Thorhallsson AT, Hunt CJ, Yang H, Morth JP, Meyer AS, Zeuner B. Structural elucidation and characterization of GH29A α-l-fucosidases and the effect of pH on their transglycosylation. FEBS J. 2025 Feb;292(3):653-680. PMID:39658312 doi:10.1111/febs.17347