糖基转移酶
大肠杆菌
代谢工程
生物合成
尿苷二磷酸
生物化学
生物催化
化学
尿苷二磷酸葡萄糖
酶
催化作用
基因
离子液体
作者
Yuncong Xu,Shiqiang Liu,Liuyun Bian,Zhenlin Li,Chen Luo,Yijun Chen,Xuri Wu
标识
DOI:10.1021/acs.jafc.1c07699
摘要
The combination of the insufficient availability and the complex structure of siamenoside I (SI), the sweetest glucoside isolated from Siraitia grosvenorii to date, limited its use as a natural sweetener. To solve this problem, an improved biocatalyst, UGT-M2, was semi-rationally created by engineering the uridine diphosphate glycosyltransferase UGT94-289-2 from S. grosvenorii for the monoglucosylation of mogroside IIIE (MG IIIE) to SI. Subsequently, an engineered Escherichia coli cell was constructed, which combined UGT-M2 with a UDP-glucose regeneration system to circumvent the need for expensive UDP-glucose to produce SI. After optimization, high-purity SI (>96.4%) was efficiently prepared from MG IIIE at a 1 L scale with a productivity of 29.78 g/(L day) and a molar yield of 76.5% and without using exogenous UDP-glucose. This study not only developed a whole-cell approach for the preparation of SI but also provided an alternative glycosyltransferase variant for SI biosynthesis with synthetic biology in the future.
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