Siamenoside Research Articles

Engineering of a UDP-Glycosyltransferase for the Efficient Whole-Cell Biosynthesis of Siamenoside I in Escherichia coli.

The combination of the insufficient availability and the complex structure of siamenoside I (SI), the sweetest glucoside isolated from Siraitia grosvenorii to date, limited its use as a natural sweetener. To solve this problem, an improved biocatalyst, UGT-M2, was semi-rationally created by engineering the uridine diphosphate glycosyltransferase UGT94-289-2 from S. grosvenorii for the monoglucosylation of mogroside IIIE (MG IIIE) to SI. Subsequently, an engineered Escherichia coli cell was constructed, which combined UGT-M2 with a UDP-glucose regeneration system to circumvent the need for expensive UDP-glucose to produce SI. After optimization, high-purity SI (>96.4%) was efficiently prepared from MG IIIE at a 1 L scale with a productivity of 29.78 g/(L day) and a molar yield of 76.5% and without using exogenous UDP-glucose. This study not only developed a whole-cell approach for the preparation of SI but also provided an alternative glycosyltransferase variant for SI biosynthesis with synthetic biology in the future.

Selective enzymatic α-1,6- monoglucosylation of mogroside IIIE for the bio-creation of α-siamenoside I, a potential high-intensity sweetener

A new compound, α-siamenoside I (α-SI), with a glucose unit selectively bound to the 6-hydroxyl group of the 24-O-β-glucosyl moiety of mogroside IIIE by α-1,6-glucosidic bond, was bio-created by two screened cyclodextrin glycosyltransferases with a maximum yield of 59.3%. Compared to mogroside IIIE, α-SI showed a significantly increased sweetness intensity (508 times sweeter than 5% sucrose), which is superior to siamenoside I (SI), the sweetest triterpenoid saponin isolated from Siraitia grosvenorii to date. Sensory evaluation showed that the taste quality of α-SI also was obviously better than mogroside IIIE. In addition to α-SI possessing a good stability similar to that of SI, it also did not cause a significant decrease in cell viability at a concentration of 200 μg/mL and had a negative influence on islets function at 1 μM. All of these preliminarily results pave the way for promoting α-SI as a potential low-calorie sweetener.