Exploration of glycosylated flavonoids from metabolically engineered <Emphasis Type="Italic">E. coli</Emphasis> |
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| Authors: | Dinesh Simkhada Nagendra Prasad Kurumbang Hei Chan Lee Jae Kyung Sohng |
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| Institution: | (1) Plant Cell Biology Millennium Nucleus, Department of Biology, Faculty of Science, University of Chile and Centre of Plant Biotechnology, University Andrés Bello, Republica 217, Santiago, Chile;(2) Present address: Department of Botany and Plant Sciences, Center for Plant Cell Biology, University of California Riverside, 2130 Batchellor Itall, Riverside, CA 92521, USA;(3) Department of Anatomy and Developmental Biology, University College London, Gower Street, London, WC1E6BT, UK; |
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| Abstract: | Flavonoids glycosylated with UDP-glucuronic acid and UDP-xylose are spatially distributed in nature. To produce these glycosides,
E. coli was engineered to overexpress biosynthetic gene clusters of UDP-sugars (galU from E. coli K12, UDP-glucose dehydrogenase (calS8), and UDP-glucuronic acid decarboxylase (calS9) from Micromonospora echinospora spp. calichensis). Flavonoids were glycosylated by overexpression of the glycosyltransferase gene (atGt-5) from Arabidopsis thaliana. Finally, metabolically engineered host E. coli (US89Gt-5) was generated. Production of flavonoid glycosides was observed in a biotransformation system consisting of flavonoids
(naringenin and quercetin) exogenously fed to host cells. The glycosylated derivatives 7-O-glucuronyl naringenin (m/z+ 449), 7-O-xylosyl naringenin (m/z+ 405), and 7-O-glucuronyl quercetin (m/z+ 479) were detected and confirmed by ESI-MS/MS, ESI-MS/MS and LC/MS-MS analysis, respectively. |
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