Identification of an α-(1,4)-Glucan-Synthesizing Amylosucrase from Cellulomonas carboniz T26.

Amylosucrase, catalyzing the synthesis of α-(1,4)-glucan from sucrose, has been widely studied and used in carbohydrate biotransformation because of its versatile activities. In this study, a novel amylosucrase was characterized from Cellulomonas carboniz T26. The recombinant enzyme was overexpressed in Escherchia coli and purified by nickel affinity chromatography. It was determined to be a monomeric protein with a molecular mass of 72 kDa. The optimum pH and temperature for transglucosylation were measured to be pH 7.0 and 40 °C. The transglucosylation activity was significantly higher than the hydrolytic activity. The main product generated from sucrose was structurally determined to be α-(1,4)-glucan. A small amount of glucose was produced by hydrolysis, and sucrose isomers including turanose and trehalulose were generated as minor products. The ratio of hydrolytic, polymerization, and isomerization reactions was calculated to be 5.8:84.0:10.2. The enzyme favored production of long-chain insoluble α-glucan at lower temperature.