Transglucosylation reaction of amylomaltase for the synthesis of anticariogenic oligosaccharides

Abstract

Abstract Synthesis of maltooligosylsucroses by the recombinant amylomaltase from Corynebacterium glutamicum as a N terminal (His)6 chimera is reported. From the analysis of the products, by TLC and HPLC analysis on a Rezex RSO-Oligosaccharide column, the suitable glucosyl donor was found to be raw tapioca starch. The optimal condition was 2.0% (w/v) sucrose, 2.5% (w/v) raw tapioca starch and 9 U/ml of amylomaltase at 30 °C for 48 h, giving an overall 82% yield of maltooligosylsucrose products. After purified by Bio-Gel P-2 size exclusion column chromatography, the main products were determined by MS and NMR analysis to be maltosyl-, maltotriosyl-, maltotretraosyl- and maltopentaosyl-fructosides (G2F, G3F, G4F and G5F, respectively, where G = glucosyl unit and F = fructose) with an α-1,4 linkage between the added glucosyl unit and the sucrose. The low cariogenic property of these maltooligosylsucrose products was confirmed by analyzing the effect on the synthesis of water insoluble glucan, acid fermentation, plaque formation and cell aggregation of Streptococcus mutans when compared to those exerted by sucrose. Moreover, by adding sucrose to maltooligosylsucrose products at ratios of 1:1, 1:2 and 1:4, the inhibitory effects on glucosyltransferase activity of S. mutans by 7, 33 and 50%, respectively, were observed. These results suggest that the obtained maltooligosylsucrose products have an anticariogenic property and could be used to substitute for sucrose in food or related products.