Zhao Man, Lai Donglian, Jia Xiaoli, Yu Mengying, Liu Zhi-Qiang, Zheng Yu-Guo
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
J Agric Food Chem. 2025 Mar 5;73(9):5341-5352. doi: 10.1021/acs.jafc.4c11011. Epub 2025 Feb 19.
Mogroside V is a triterpene, a natural high-intensity sweetener, isolated from the fruits of . Selective glycosylation of mogrol is a feasible approach for the biosynthesis of mogroside V. In this study, glycosyltransferase UGTM1 and UGTM2 were engineered to UGTM1-3 and UGTM2-4, which selectively and directly transfer glucose from UDPG to 3'-hydroxyl and 24'-hydroxyl groups and their branch chains of the mogrol moiety for the biosynthesis of mogroside V. The enzyme activities of UGTM1-3 and UGTM2-4 were enhanced 2.88 and 3.60 times, respectively. To eliminate the need for UDPG and improve productivity, a UDPG regeneration system was introduced to couple with the UGTs. Finally, mogrol was directly converted to mogroside V by UGTM1-3, UGTM2-4, and AtSUS1 with a conversion rate of 18.2% without the exogenous addition of UDPG. This study provides an in vitro multienzyme cascade catalytic system for the efficient conversion of mogroside V.
罗汉果甜苷V是一种三萜类化合物，是从罗汉果果实中分离得到的天然高强度甜味剂。罗汉果甜苷元的选择性糖基化是罗汉果甜苷V生物合成的一种可行方法。在本研究中，将糖基转移酶UGTM1和UGTM2改造为UGTM1-3和UGTM2-4，它们能将UDPG中的葡萄糖选择性地直接转移到罗汉果甜苷元部分的3'-羟基和24'-羟基及其支链上，用于罗汉果甜苷V的生物合成。UGTM1-3和UGTM2-4的酶活性分别提高了2.88倍和3.60倍。为了消除对UDPG的需求并提高生产率，引入了UDPG再生系统与UGTs偶联。最后，在不额外添加UDPG的情况下，UGTM1-3、UGTM2-4和AtSUS1将罗汉果甜苷元直接转化为罗汉果甜苷V，转化率为18.2%。本研究为罗汉果甜苷V的高效转化提供了一种体外多酶级联催化系统。
