Semi-rational engineering of an α-L-fucosidase for regioselective synthesis of fucosyl-<i>N</i>-acetylglucosamine disaccharides.

ABSTRACT: α-L-Fucosidases are attractive biocatalysts for the production of bioactive fucosylated oligosaccharides, however, poor regioselectivity and activity for transglycosylation have significantly limited their applications. We have recently derived an α-L-Fucosidase, BF3242, from Bacteroides fragilis NCTC9343, which could efficiently synthesize a mixture of Fuc-α-1,3/1,6-GlcNAc, but its 1,3/1,6-regioselectivity was observably affected by reaction temperature. Here, we integrated loop-targeted random mutagenesis and site-directed mutagenesis to engineer the regioselectivity and transglycosylation activity of BF3242. Loop-targeted random mutagenesis revealed that L266 in the loop-4 (H242-S267) within the model of BF3242 was a key residue for the regioselectivity for transglycosylation, an

SUBMITTER: Liu P 

PROVIDER: S-EPMC11875152 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature