335193-88-9Relevant articles and documents
On expanding the repertoire of glycosynthases: Mutant β-galactosidases forming β-(1,6)-linkages
Jakeman, David L.,Withers, Stephen G.
, p. 866 - 870 (2002)
Oligosaccharide synthesis by enzymatic processes offers the potential for thrusting oligosaccharides to the forefront of pharmaceutical research, in part, due to expedient and scalable reaction protocols. Glycosynthases are an emerging class of mutant enzymes capable of synthesizing glycosidic linkages in high yield. We report a new glycosynthase enzyme generated by a point mutation of E. coli β-galactosidase that condenses α-galactosyl fluoride with aryl glucosides forming a β-(1,6) glycosidic linkage. A further point mutation within the enzyme, proximal to the active site, increases the yields significantly.
Characterization of properties and transglycosylation abilities of recombinant α-galactosidase from cold-adapted marine bacterium pseudoalteromonas KMM 701 and its C494N and D451A mutants
Bakunina, Irina,Slepchenko, Lubov,Anastyuk, Stanislav,Isakov, Vladimir,Likhatskaya, Galina,Kim, Natalya,Tekutyeva, Liudmila,Son, Oksana,Balabanova, Larissa
, (2018/10/20)
A novel wild-type recombinant cold-active α-D-galactosidase (α-PsGal) from the cold-adapted marine bacterium Pseudoalteromonas sp. KMM 701, and its mutants D451A and C494N, were studied in terms of their structural, physicochemical, and catalytic properties. Homology models of the three-dimensional α-PsGal structure, its active center, and complexes with D-galactose were constructed for identification of functionally important amino acid residues in the active site of the enzyme, using the crystal structure of the α-galactosidase from Lactobacillus acidophilus as a template. The circular dichroism spectra of the wild α-PsGal and mutant C494N were approximately identical. The C494N mutation decreased the efficiency of retaining the affinity of the enzyme to standard p-nitrophenyl-α-galactopiranoside (pNP-α-Gal). Thin-layer chromatography, matrix-assisted laser desorption/ionization mass spectrometry, and nuclear magnetic resonance spectroscopy methods were used to identify transglycosylation products in reaction mixtures. α-PsGal possessed a narrow acceptor specificity. Fructose, xylose, fucose, and glucose were inactive as acceptors in the transglycosylation reaction. α-PsGal synthesized -α(1→6)- and -α(1→4)-linked galactobiosides from melibiose as well as -α(1→6)- and -α(1→3)-linked p-nitrophenyl-digalactosides (Gal2-pNP) from pNP-α-Gal. The D451A mutation in the active center completely inactivated the enzyme. However, the substitution of C494N discontinued the Gal-α(1→3)-Gal-pNP synthesis and increased the Gal-α(1→4)-Gal yield compared to Gal-α(1→6)-Gal-pNP.
Preparation of α-galactooligoglycosides by cell walls from Cryptococcus laurentii using a novel α-galactosyl donor
Mastihuba, Vladimír,Mastihubová, Mária,Belák, Miroslav,Dudíková, Jana,Potocká, Elena Karni?ová,Petru?, Ladislav
, p. 1089 - 1094 (2017/10/05)
The cell walls of an acapsular strain of the yeast Cryptococcus laurentii catalyze the regioselective formation of α-galactooligosaccharides through self-condensation of 4-nitrophenyl α-D-galactopyranoside and of a novel activated α-galactosyl donor 2,2,2-trifluoroethyl α-D-galactopyranoside. The latter substance can be easily prepared by several methods and is highly soluble in water and therefore can be used in higher initial concentrations suppressing secondary product hydrolysis. The preparative reaction catalyzed by cell walls provided 17.4% and 2% of corresponding 2,2,2-trifluoroethyl galactobioside and galactotrioside, respectively, while the reaction with 4-nitrophenyl α-D-galactopyranoside provided the corresponding 4-nitrophenyl galactobioside and galactotrioside in 6.6 and 2.5% yields, respectively. The reactions proceeded with strict α-(1 → 6)-regioselectivity.
Acceptor-induced modification of regioselectivity in CGTase-catalyzed glycosylations of p-nitrophenyl-glucopyranosides
Strompen, Simon,Miranda-Molina, Alfonso,López-Munguía, Agustín,Castillo, Edmundo,Saab-Rincón, Gloria
, p. 46 - 54 (2015/03/05)
Cyclodextrin glycosyltransferases (CGTase) are reported to selectively catalyze α(1→4)-glycosyl transfer reactions besides showing low hydrolytic activity. Here, the effect of the anomeric configuration of the glycosyl acceptor on the regioselectivity of
