Highly Efficient Production of Rebaudioside D Enabled by Structure-guided Engineering of Bacterial Glycosyltransferase YojK

Overview

Journal Front Bioeng Biotechnol

Date 2022 Sep 12

PMID 36091437

Authors

Baodang Guo

Xiaodong Hou

Yan Zhang

Zhiwei Deng

Qian Ping

Kai Fu

Zhenbo Yuan

Yijian Rao

Affiliations

Soon will be listed here.

Abstract

Owing to zero-calorie, high-intensity sweetness and good taste profile, the plant-derived sweetener rebaudioside D (Reb D) has attracted great interest to replace sugars. However, low content of Reb D in Bertoni as well as low soluble expression and enzymatic activity of plant-derived glycosyltransferase in Reb D preparation restrict its commercial usage. To address these problems, a novel glycosyltransferase YojK from 168 with the ability to glycosylate Reb A to produce Reb D was identified. Then, structure-guided engineering was performed after solving its crystal structure. A variant YojK-I241T/G327N with 7.35-fold increase of the catalytic activity was obtained, which allowed to produce Reb D on a scale preparation with a great yield of 91.29%. Moreover, based on the results from molecular docking and molecular dynamics simulations, the improvement of enzymatic activity of YojK-I241T/G327N was ascribed to the formation of new hydrogen bonds between the enzyme and substrate or uridine diphosphate glucose. Therefore, this study provides an engineered bacterial glycosyltransferase YojK-I241T/G327N with high solubility and catalytic efficiency for potential industrial scale-production of Reb D.

Citing Articles

Seeds of Bertoni as a Source of Plant Growth-Promoting Endophytic Bacteria with the Potential to Synthesize Rebaudioside A.

Simlat M, Ptak A, Jaglarz A, Szewczyk A, Dziurka M, Gurgul A Int J Mol Sci. 2023; 24(3).

PMID: 36768498 PMC: 9917351. DOI: 10.3390/ijms24032174.

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