Novel enzymatic production of trehalose from sucrose using amylosucrase and maltooligosyltrehalose synthase-trehalohydrolase

Abstract

Trehalose (a-D-glucopyranosyl a-D-glucopyranoside) is an important non-reducing disaccharide used in the food industry due to its mild sweetness (45% that of sucrose), low cariogenicity, high glass transition temperature, low hygroscopicity, and protein protection properties. In this study, we accomplished the production of trehalose from sucrose as a sole substrate using a novel dual-enzyme system, in which amylosucrase (ASase) and maltooligosyltrehalose synthase-trehalohydrolase (MTSH) fusion enzyme were employed. The biotransformation of sucrose to trehalose was confirmed by high-performance anionexchange chromatography (HPAEC) analysis. Trehalose was successfully produced by both simultaneous and sequential methods by using ASase and MTSH. A higher trehalose production yield (3.15 ± 0.83 mM trehalose/ 20 mM sucrose) was observed in the sequential method than the simultaneous method (1.43 ± 0.14 mM trehalose/ 20 mM sucrose), indicating that the production of maltooligosaccharides from sucrose by ASase was an important step in the biosynthesis of trehalose.

Glycoside Hydrolases and Glycosyltransferases from Hyperthermophilic Archaea: Insights on Their Characteristics and Applications in Biotechnology

“…Trehalose synthases are retaining GTs that catalyze the formation of trehalose, by transferring a glucose from NDP-glucose to another glucose residue. The resulting product is a non-reducing disaccharide that can be used either as a food additive because it has lower sweetness than sucrose, a moderate glycemic index and low cariogenicity, or can serve as a stabilizer in cosmetics and pharmaceuticals due to its high heat and pH resistance, high glass transition temperature, high water retention capacity and protective effect on proteins [160][161][162].…”

Expression of Deinococcus geothermalis trehalose synthase gene in Escherichia coli and its enzymatic properties

“…Then, maltooligosyltrehalose trehalohydrolase (MTHase) cleaves the -1,4-glucosidic linkage next to the -1,1linkage of trehalosyl-oligosaccharide to produce trehalose and oligosaccharide with lower molecular mass (Kato, 1999). Recently, Kim et al (2011) reported two steps enzymatic production of trehalose from sucrose using amylosucrose from D. geothermalis and maltooligosyltrehalose synthase from Bacillus helvolum. The third pathway, catalyzed by trehalose glycosyltransferring synthase (TreT), involves the conversion of nucleoside diphosphate glucose and glucose, instead of glucose-6phosphate, into trehalose (Ryu et al, 2005;Empadinhas and da Costa, 2006).…”

Cascade Biocatalytic Processes through Combined Cross-linked Enzyme Aggregates

“…Among several synthetic strategies studied for the synthesis of trehalose, the biocatalytic one starting from sucrose has aroused considerable interest [60]. The biocatalysts involved in this process are amylosucrase (ASase), maltooligosyl trehalose synthase (MTS) and maltooligosyltrehalose trealohydrolase (MTH).…”