|Content:||10 Units at 25oC|
|Formulation:||In 3.2 M ammonium sulphate|
|Stability:||> 4 years at 4oC|
|Synonyms:||alpha-L-fucosidase; alpha-L-fucoside fucohydrolase|
|Concentration:||Supplied at ~ 5 U/mL|
|Expression:||Recombinant from Thermotoga maritima|
|Specificity:||Hydrolysis of α-L-fucose residues from the non-reducing terminal residues in glycoproteins and oligosaccharides.|
|Specific Activity:|| ~ 2.5 U/mg protein (25oC); |
~ 4 U/mg protein (37oC);
~ 15 U/mg protein (60oC);
~ 200 U/mg protein (95oC)
|Unit Definition:||One Unit of α-fucosidase activity is defined as the amount of enzyme required to release one µmole of p-nitrophenol (pNP) per minute from p-nitrophenyl-α-L-fucopyranoside in citrate buffer (50 mM)/phosphate buffer (100 mM), pH 5.0.|
|Application examples:||For use in glycobiology research.|
Root-knot nematode chemotaxis is positively regulated by L-galactose sidechains of mucilage carbohydrate rhamnogalacturonan-I.
Tsai, A. Y. L., Iwamoto, Y., Tsumuraya, Y., Oota, M., Konishi, T., Ito, S., Kotake, T., Ishikawa, H. & Sawa, S. (2021). Science Advances, 7(27), eabh4182.
Root-knot nematodes (RKNs) are plant parasites and major agricultural pests. RKNs are thought to locate hosts through chemotaxis by sensing host-secreted chemoattractants; however, the structures and properties of these attractants are not well understood. Here, we describe a previously unknown RKN attractant from flaxseed mucilage that enhances infection of Arabidopsis and tomato, which resembles the pectic polysaccharide rhamnogalacturonan-I (RG-I). Fucose and galactose sidechains of the purified attractant were found to be required for attractant activity. Furthermore, the disaccharide α-L-galactosyl-1,3-L-rhamnose, which forms the linkage between the RG-I backbone and galactose sidechains of the purified attractant, was sufficient to attract RKN. These results show that the α-L-galactosyl-1,3-L-rhamnose linkage in the purified attractant from flaxseed mucilage is essential for RKN attraction. The present work also suggests that nematodes can detect environmental chemicals with high specificity, such as the presence of chiral centers and hydroxyl groups.Hide Abstract
Development of a 1, 2-difluorofucoside activity-based probe for profiling GH29 fucosidases.
Luijkx, Y. M., Jongkees, S., Strijbis, K. & Wennekes, T. (2021). Organic & Biomolecular Chemistry, 19(13), 2968-2977.
GH29 α-L-fucosidases catalyze hydrolysis of terminal α-L-fucosyl linkages with varying specificity and are expressed by prominent members of the human gut microbiota. Both homeostasis and dysbiosis at the human intestinal microbiota interface have been correlated with altered fucosidase activity. Herein we describe the development of a 2-deoxy-2-fluoro fucosyl fluoride derivative with an azide mini-tag as an activity-based probe (ABP) for selective in vitro labelling of GH29 α-L-fucosidases. Only catalytically active fucosidases are inactivated by this ABP, allowing their functionalization with a biotin reporter group via the CuAAC reaction and subsequent in-gel detection at nanogram levels. The ABP we present here is shown to be active against a GH29 α-L-fucosidase from Bacteroides fragilis and capable of labeling two other GH29 α-L-fucosidases with different linkage specificity, illustrating its broader utility. This novel ABP is a valuable addition to the toolbox of fucosidase probes by allowing identification and functional studies of the wide variety of GH29 fucosidases, including those in the gut microbiota.Hide Abstract
Guzmán-Rodríguez, F., Alatorre-Santamaría, S., Gómez-Ruiz, L., Rodríguez-Serrano, G., García-Garibay, M. & Cruz-Guerrero, A. (2018). Extremophiles, 1-6.
The influence of CaCl2 and NaCl in the hydrolytic activity and the influence of CaCl2 in the synthesis of fucosylated oligosaccharides using α-L-fucosidase from Thermotoga maritima were evaluated. The hydrolytic activity of α-L-fucosidase from Thermotoga maritima displayed a maximum increase of 67% in the presence of 0.8 M NaCl with water activity (aw) of 0.9672 and of 138% in the presence of 1.1 M CaCl2 (aw 0.9581). In addition, the hydrolytic activity was higher when using CaCl2 compared to NaCl at aw of 0.8956, 0.9581 and 0.9672. On the other hand, the effect of CaCl2 in the synthesis of fucosylated oligosaccharides using 4-nitrophenyl-fucose as donor substrate and lactose as acceptor was studied. In these reactions, the presence of 1.1 M CaCl2 favored the rate of transfucosylation, and improved the yield of synthesis duplicating and triplicating it with lactose concentrations of 58 and 146 mM, respectively. CaCl2 did not significatively affect hydrolysis rate in these reactions. The combination of the activating effect of CaCl2, the decrement in aw and lactose concentration had a synergistic effect favoring the synthesis of fucosylated oligosaccharides.Hide Abstract