Journal
SCIENTIFIC REPORTS
Volume 10, Issue 1, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41598-020-63379-9
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Funding
- National Key R&D Program of China [2018YFA0902000]
- National Natural Science Foundation of China [21778076]
- 111 Project from the Ministry of Education of China
- State Administration of Foreign Expert Affairs of China [111-2-07]
- Double First-Class University project [CPU2018GY36]
- Postgraduate Research & Practice Innovation Program of Jiangsu Province [KYCX19_0659]
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Rebaudioside KA is a diterpene natural sweetener isolated in a trace amount from the leaves of Stevia 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 a beta-1,2 linkage at 19-COO-beta-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 engineered Escherichia coli strain to couple with the glycosyltransferase. Finally, rubusoside at 22.5g/L (35.0mM) 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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