Biosynthesis of flavone C-glucosides in engineered Escherichia coli.

Applied Microbiology and Biotechnology
Anil ShresthaJae Kyung Sohng

Abstract

Two plant-originated C-glucosyltransferases (CGTs) UGT708D1 from Glycine max and GtUF6CGT1 from Gentiana triflora were accessed for glucosylation of selected flavones chrysin and luteolin. Uridine diphosphate (UDP)-glucose pool was enhanced in Escherichia coli cell cytosol by introducing heterologous UDP-glucose biosynthetic genes, i.e., glucokinase (glk), phosphoglucomutase (pgm2), and glucose 1-phosphate uridylyltransferase (galU), along with glucose facilitator diffusion protein from (glf) from different organisms, in a multi-monocistronic vector with individual T7 promoter, ribosome binding site, and terminator for each gene. The C-glucosylated products were analyzed by high-performance liquid chromatography-photodiode array, high-resolution quadruple time-of-flight electrospray ionization mass spectrometry, and one-dimensional nuclear magnetic resonance analyses. Fed-batch shake flask culture showed 8% (7 mg/L; 16 μM) and 11% (9 mg/L; 22 μM) conversion of chrysin to chrysin 6-C-β-D-glucoside with UGT708D1 and GtUF6CGT1, respectively. Moreover, the bioengineered E. coli strains with exogenous UDP-glucose biosynthetic genes and glucose facilitator diffusion protein enhanced the production of chrysin 6-C-β-D-glucoside by appr...Continue Reading

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Citations

Jan 19, 2019·Microbial Cell Factories·Biplav ShresthaJae Kyung Sohng
Jun 10, 2020·Applied Microbiology and Biotechnology·Yuan JiXing Zhang
Sep 5, 2018·Applied Microbiology and Biotechnology·Anil ShresthaJae Kyung Sohng
Oct 26, 2018·Emerging Topics in Life Sciences·Ramesh Prasad PandeyJae Kyung Sohng
Apr 3, 2021·Journal of the American Chemical Society·Dongsoo YangSang Yup Lee
Aug 8, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Hanghang LouQihe Chen

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Methods Mentioned

BETA
glycosylation
PCR
electrophoresis
glucosylation
NMR
C
-glucosylation

Software Mentioned

TopSpin
MestReNova

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