Selective synthesis of rebaudioside M2 through structure-guided engineering of glycosyltransferase UGT94D1.

ABSTRACT: Rebaudioside M2 (Reb M2), a novel steviol glycoside derivative, has limited industrial applications due to its low synthetic yield and selectivity. Herein, we identify UGT94D1 as a selective glycosyltransferase for rebaudioside D (Reb D), leading to the production of a mono β-1,6-glycosylated derivative, Reb M2. A variant UGT94D1-F119I/D188P was developed through protein engineering. This mutant exhibited a 6.33-fold improvement in catalytic efficiency, and produced Reb M2 with 92% yield. Moreover, molecular dynamics simulations demonstrated that UGT94D1-F119I/D188P exhibited a shorter distance between the nucleophilic oxygen (OH6) of the substrate Reb D and uridine diphosphate glucose, along with an increased O^phosphate-C1-O^acceptor angle, thus improving the c

SUBMITTER: Yang L 

PROVIDER: S-EPMC10875103 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature