Enhancement of Rebaudioside M Production by Structure-Guided Engineering of Glycosyltransferase UGT76G1
Baodang Guo, Zhiwei Deng, Fei Meng, Qingfu Wang, Yan Zhang, Zhenbo Yuan +1 more
Journal of Agricultural and Food Chemistry
Abstract
Owing to zero-calorie and advanced organoleptic properties similar to sucrose, the plant-derived rebaudioside M (Reb M) has been considered as a next generation sweetener. However, a low content of Reb M in <i>Stevia rebaudiana</i> Bertoni and low enzymatic activity of UGT76G1, which is an uridine diphosphate glucose (UDPG)-dependent glycosyltransferase with the ability to glycosylate rebaudioside D (Reb D) to produce Reb M through the formation of β-1,3 glycosidic bond, restrict its commercial usage. To improve the catalytic activity of UGT76G1, a variant UGT76G1-T284S/M88L/L200A was obtained by structure-guided evolution, whose catalytic activity toward Reb D increased by 2.38 times compared with UGT76G1-T284S. This allowed us to prepare Reb M on a large-scale with a great yield of 90.50%. Moreover, molecular dynamics simulation illustrated that UGT76G1-T284S/M88L/L200A reduced distances from Reb D to catalytic residues and UDPG. Hence, we report an efficient method for the potential scale production of Reb M in this study.
Extracted Claims
4 claims extracted from this paper into the knowledge graph
rebaudioside M (Reb M) considered next generation sweetener
“Owing to zero-calorie and advanced organoleptic properties similar to sucrose, the plant-derived rebaudioside M (Reb M) has been considered as a next generation sweetener.”
UGT76G1-T284S/M88L/L200A reduced distances from Reb D to catalytic residues and UDPG
“Moreover, molecular dynamics simulation illustrated that UGT76G1-T284S/M88L/L200A reduced distances from Reb D to catalytic residues and UDPG.”
UGT76G1 glycosylates rebaudioside D (Reb D) to produce Reb M
“UGT76G1, which is an uridine diphosphate glucose (UDPG)-dependent glycosyltransferase with the ability to glycosylate rebaudioside D (Reb D) to produce Reb M through the formation of β-1,3 glycosidic ...”