7VM0

Crystal structure of YojK from B.subtilis in complex with UDP

7VM0 の概要

• エントリーDOI: 10.2210/pdb7vm0/pdb
• 分子名称: Glycosyl transferase family 1, URIDINE-5'-DIPHOSPHATE, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: complex, dimer, transferase
• 由来する生物種: Bacillus subtilis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 94806.65

構造登録者

Hou, X.D.,Guo, B.D.,Rao, Y.J. (登録日: 2021-10-06, 公開日: 2022-10-12, 最終更新日: 2023-11-29)

主引用文献

Guo, B.,Hou, X.,Zhang, Y.,Deng, Z.,Ping, Q.,Fu, K.,Yuan, Z.,Rao, Y.
Highly efficient production of rebaudioside D enabled by structure-guided engineering of bacterial glycosyltransferase YojK.
Front Bioeng Biotechnol, 10:985826-985826, 2022

PubMed 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.

PubMed: 36091437
DOI: 10.3389/fbioe.2022.985826

実験手法

X-RAY DIFFRACTION (1.9 Å)