Efficient Biocatalytic Preparation of Rebaudioside KA: Highly Selective Glycosylation Coupled with UDPG Regeneration

Efficient Biocatalytic Preparation of Rebaudioside KA: Highly Selective Glycosylation Coupled with UDPG Regeneration
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莱鲍迪甙 KA 的高效生物催化制备:高选择性糖基化结合 UDPG 再生

DOI:
10.1038/s41598-020-63379-9
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发表时间:
2020-04
期刊:
影响因子:
4.6
通讯作者:
Wu Xuri
Wu Xuri
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Zhang Yunyun;Xu Shaohua;Jin Yue;Dai Yan;Chen Yijun;Wu Xuri

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甜菊糖苷KA是一种从甜菊叶中分离出的微量二萜天然甜味剂。雷鲍迪糖苷是一种丰富存在于多种植物中的天然产物,其选择性糖基化是生物合成雷鲍迪糖苷KA的可行途径。在本研究中,发现细菌糖基转移酶OleD选择性地将葡萄糖从UDPG转移到rubuso苷的19-COO-β- d -葡萄糖基上具有a -β- 1,2连锁的2 ' -羟基上,用于生物合成rebodio苷KA。为了消除UDPG的使用,提高生产效率,构建了一个UDPG再生系统,作为工程大肠杆菌与糖基转移酶偶联的大肠杆菌。最后,在不外源添加UDPG的情况下,22.5 g/L (35.0 mM)的鲁布迪糖苷被全细胞完全转化为雷鲍迪糖苷KA。本研究为雷鲍迪甙KA的高选择性生物合成提供了一种高效、可扩展的方法。
Rebaudioside KA is a diterpene natural sweetener isolated in a trace amount from the leaves ofStevia rebaudiana. Selective glycosylation of rubusoside, a natural product abundantly presented in various plants, is a feasible approach for the biosynthesis of rebaudioside KA. In this study, bacterial glycosyltransferase OleD was identified to selectively transfer glucose from UDPG to 2′-hydroxyl group with aβ-1,2 linkage at 19-COO-β-D-glucosyl moiety of rubusoside for the biosynthesis of rebaudioside KA. To eliminate the use of UDPG and improve the productivity, a UDPG regeneration system was constructed as an engineeredEscherichia colistrain to couple with the glycosyltransferase. Finally, rubusoside at 22.5 g/L (35.0 mM) was completely converted to rebaudioside KA by the whole cells without exogenous addition of UDPG. This study provides an efficient and scalable method for highly selective biosynthesis of rebaudioside KA.
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