Academic Journal
Transglycosylation Capabilities of Wild-Type α-l-Fucosidase iso1 from Paenibacillus thiaminolyticus and Its Engineered Mutants: Preparation of Fucosylated Oligosaccharides.
| Τίτλος: | Transglycosylation Capabilities of Wild-Type α-l-Fucosidase iso1 from Paenibacillus thiaminolyticus and Its Engineered Mutants: Preparation of Fucosylated Oligosaccharides. |
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| Συγγραφείς: | Vodičková P; Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic., Klimešová L; Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic.; Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic., Nekvasilová P; Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic.; Department of Genetics and Microbiology, Faculty of Science, Charles University, Prague, Czech Republic., Petrásková L; Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic., Pelantová H; Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic., Kovaľová T; Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic., Lipovová P; Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic., Bojarová P; Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic., Benešová E; Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Czech Republic. |
| Πηγή: | Microbial biotechnology [Microb Biotechnol] 2026 Apr; Vol. 19 (4), pp. e70354. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 101316335 Publication Model: Print Cited Medium: Internet ISSN: 1751-7915 (Electronic) Linking ISSN: 17517915 NLM ISO Abbreviation: Microb Biotechnol Subsets: MEDLINE |
| Imprint Name(s): | Publication: Hoboken, NJ : Wiley-Blackwell Original Publication: Oxford : Blackwell, c2008- |
| Ιατρικοί όροι (MeSH): | Paenibacillus*/enzymology , Paenibacillus*/genetics , alpha-L-Fucosidase*/genetics , alpha-L-Fucosidase*/metabolism , alpha-L-Fucosidase*/chemistry , Fucose*/metabolism , Oligosaccharides*/metabolism , Oligosaccharides*/chemistry, Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Mutant Proteins/metabolism ; Mutant Proteins/genetics ; Mutagenesis, Site-Directed ; Glycosylation |
| Περίληψη: | To enhance the yield of l-fucosylated molecules synthesized via transfucosylation, we employed α-l-transfucosidases, enzymes engineered from α-l-fucosidases to favour transglycosylation over hydrolysis. This study investigated the transglycosylation potential of mutated variants of α-l-fucosidase iso1 from Paenibacillus thiaminolyticus designed by structural comparison with Thermotoga maritima α-l-transfucosidase. Using site-directed mutagenesis, point mutations (S237A, S237G, S237P, S237V, Y189F) were introduced, and the corresponding recombinant proteins were successfully expressed and purified. Among them, the S237V variant achieved the highest overall yield of fucosylated lactose and increased the transglycosylation/hydrolysis ratio more than 20-fold compared to the wild-type enzyme. This variant enabled regioselective synthesis of 3'-and 6'-fucosyllactose, as well as a functionalized 3'-fucosyllactose bearing a Boc-protected ethylthioureidyl linker (3'-FucLac-tBoc), with all structures confirmed by NMR spectroscopy. The engineered α-l-transfucosidase iso1 from Paenibacillus thiaminolyticus catalysed the regioselective synthesis of structurally defined fucosyllactose derivatives, including C-1 functionalized analogs suitable for immobilization on biosensor surfaces or macromolecular scaffolds, thus expanding the biocatalytic toolbox and demonstrating the potential of semi-rational enzyme design for targeted HMO-based glycoengineering applications. (© 2026 The Author(s). Microbial Biotechnology published by John Wiley & Sons Ltd.) |
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| Grant Information: | LUC24024 Ministerstvo Školství, Mládeže a Tělovýchovy; 25-16301K Grantová Agentura České Republiky |
| Contributed Indexing: | Keywords: Paenibacillus thiaminolyticus; fucosidase; fucosyllactose; human milk oligosaccharide; site‐directed mutagenesis; transglycosidase; transglycosylation |
| Substance Nomenclature: | EC 3.2.1.51 (alpha-L-Fucosidase) 28RYY2IV3F (Fucose) 0 (Oligosaccharides) 0 (Recombinant Proteins) 0 (Mutant Proteins) |
| Entry Date(s): | Date Created: 20260423 Date Completed: 20260715 Latest Revision: 20260813 |
| Update Code: | 20260813 |
| PubMed Central ID: | PMC13103868 |
| DOI: | 10.1111/1751-7915.70354 |
| PMID: | 42023498 |
| Βάση Δεδομένων: | MEDLINE |
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