How is Trehalulose Formed by Australian Stingless Bees? - An Intermolecular Displacement of Nectar Sucrose
Jiali Zhang, Natasha L. Hungerford, Hans S. A. Yates, Tobias J. Smith, Mary T. Fletcher
Abstract
Trehalulose, a rare sucrose isomer, is a dominant sugar in stingless bee honey, with traces of the trisaccharide erlose. Incubating sucrose solutions with macerated stingless bee parts (head, thorax, and abdomen) from Tetragonula carbonaria, we observed that sucrose isomerization occurs predominantly in the head incubations, with trehalulose constituting 76.2–80.0% of total detected sugar. By contrast, sucrose hydrolysis occurred in stingless bee abdomen incubations, with glucose and fructose observed as 48.6–51.7% and 48.3–49.7%, respectively, of total detected sugar. Incubating glucose/fructose (1:1) solutions with any bee part did not result in trehalulose formation. In addition, by tracing the 13C isotope-labeled monosaccharide moieties throughout the isomerization from sucrose to trehalulose and erlose, for the first time, the mechanism was established as an enzymatic double displacement reaction. Sucrose acts as a glucose donor giving a β-d-glucosyl enzyme intermediate with fructose release as demonstrated by mixed isotope products. Glucosylation of fructose (inter- or intramolecularly) with isomerization forms trehalulose (favorable), while glucosylation of sucrose forms erlose (less favorable).