Glycosyltransferase engineering and multi-glycosylation routes development facilitating synthesis of high-intensity sweetener mogrosides

Jiao Li; Shicheng Mu; Jiangang Yang; Cui Liu; Yanfei Zhang; Peng Chen; Yan Zeng; Yueming Zhu; Yuanxia Sun

doi:10.1016/j.isci.2022.105222

Glycosyltransferase engineering and multi-glycosylation routes development facilitating synthesis of high-intensity sweetener mogrosides

iScience. 2022 Sep 27;25(10):105222. doi: 10.1016/j.isci.2022.105222. eCollection 2022 Oct 21.

Authors

Jiao Li^{1

2}, Shicheng Mu¹, Jiangang Yang¹, Cui Liu¹, Yanfei Zhang¹, Peng Chen¹, Yan Zeng¹, Yueming Zhu¹, Yuanxia Sun^{1

2}

Affiliations

¹ National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 Xi Qi Dao, Tianjin Airport Economic Area, Tianjin 300308, China.
² National Technology Innovation Center of Synthetic Biology, Tianjin 300308, China.

Abstract

Mogrosides are widely served as natural zero-calorie sweeteners. To date, the biosynthesis of high-intensity sweetness mogrosides V from mogrol has not been achieved because of inefficient and uncontrollable multi-glycosylation process. To address this challenge, we reported three UDP-glycosyltransferases (UGTs) catalyzing the primary and branched glycosylation of mogrosides and increased the catalytic efficiency by 74-400-folds toward branched glycosylation using an activity-based sequence conservative analysis engineering strategy. The computational studies provided insights into the origin of improved catalytic activity. By virtue of UGT mutants, we provided regio- and bond-controllable multi-glycosylation routes, successfully facilitating sequential glycosylation of mogrol to three kinds of mogroside V in excellent yield of 91-99%. Meanwhile, the feasibility of the routes was confirmed in engineered yeasts. It suggested that the multi-glycosylation routes would be combined with mogrol synthetic pathway to de novo produce mogrosides from glucose by aid of metabolic engineering and synthetic biology strategies in the future.

Keywords: Biosynthesis; Biotechnology; Chemistry.