Synthesis of a Fucosylated Trisaccharide Via Transglycosylation by α-l-Fucosidase from Thermotoga maritima
| Autor: | Lorena Gómez-Ruiz, Alma Cruz-Guerrero, Gabriela Rodríguez-Serrano, Mariano García-Garibay, Sergio Alatorre-Santamaría, Francisco Guzmán-Rodríguez |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Glycosylation Stereochemistry Lactose Bioengineering Applied Microbiology and Biotechnology Biochemistry High-performance liquid chromatography Substrate Specificity 03 medical and health sciences Hydrolysis chemistry.chemical_compound Thermotoga maritima Trisaccharide Fucosidase Molecular Biology Chromatography High Pressure Liquid Fucose alpha-L-Fucosidase chemistry.chemical_classification 030102 biochemistry & molecular biology biology Substrate (chemistry) General Medicine Oligosaccharide biology.organism_classification 030104 developmental biology chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Trisaccharides Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 186:681-691 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | Fucosylated oligosaccharides, such as 2'-fucosyllactose in human milk, have important biological functions such as prebiotics and preventing infection. In this work, the effect of an acceptor substrate (lactose) and the donor substrate 4-nitrophenyl-α-L-fucopyranoside (pNP-Fuc) on the synthesis of a fucosylated trisaccharide was studied in a transglycosylation reaction using α-L-fucosidase from Thermotoga maritima. Conducting a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), it was demonstrated that synthesized oligosaccharide corresponded to a fucosylated trisaccharide, and high-performance liquid chromatography (HPLC) of the hydrolyzed compound confirmed it was fucosyllactose. As the concentration of the acceptor substrate increased, the concentration and synthesis rate of the fucosylated trisaccharide also increased, and the highest concentration obtained was 0.883 mM (25.2% yield) when using the higher initial lactose concentration (584 mM). Furthermore, the lower donor/acceptor ratio had the highest synthesis, so at the molar ratio of 0.001, a concentration of 0.286 mM was obtained (32.5% yield). |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink