Production and use of glucosyltransferases from Leuconostoc mesenteroides NRRL B-1299 for the synthesis of oligosaccharides containing alpha-(1-->2) linkages.
Glucosyltransferase activities, produced by batch culture of Leuconostoc mesenteroides NRRL B-1299, were recovered both in the culture supernatant (SGT) and associated with the insoluble part of the culture (IGT). A total glucosyltransferase activity of 3.5 U/mL was measured in batch culture. The enzymes from the supernatant were purified 313 times using aqueous two-phase partition between dextran and PEG phases, yielding a preparation with 18.8 U/mg protein. It was shown that both SGT and IGT preparations catalyze acceptor reactions and transfer the glucose unit from sucrose onto maltose to produce glucooligosaccharides. Some of the glucooligosaccharides synthesized (Ln series) contain alpha-(1-->6) osidic linkages and a maltose residue at the reducing end. They were completely hydrolyzed by glucoamylase and dextranase. The other glucooligosaccharides synthesized (Bn series) resisted the action of these enzymes. The tetrasaccharide of this series has been characterized by 13C NMR. Its structure was determined as 2-O-alpha-D-glucosylpanose. The oligosaccharides synthesized by the maltose acceptor reaction with the SGT and IGT preparations only differed in the relative amounts in which they were produced. The difference may arise from diffusional limitations appearing when the insoluble catalyst is used. Under the assay conditions, the glucanase resistant oligosaccharide yield was 35% with both glucosyltransferase preparations.
IUT Biologie Appliquée, Auch, France.
M
M
Remaud-Simeon
A
A
Lopez-Munguia
V
V
Pelenc
F
F
Paul
P
P
Monsan
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002384:Catalysis; D005964:Glucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D007934:Leuconostoc / Q000201:enzymology*; D008320:Maltose / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry; D012995:Solubility
D009844:Oligosaccharides; D008320:Maltose; D005964:Glucosyltransferases