Applied Microbiology and Biotechnology

, Volume 97, Issue 5, pp 1889–1901 | Cite as

Production of 3-O-xylosyl quercetin in Escherichia coli

  • Ramesh Prasad Pandey
  • Sailesh Malla
  • Dinesh Simkhada
  • Byung-Gee Kim
  • Jae Kyung Sohng
Biotechnological products and process engineering

Abstract

Quercetin, a flavonol aglycone, is one of the most abundant flavonoids with high medicinal value. The bioavailability and pharmacokinetic properties of quercetin are influenced by the type of sugars attached to the molecule. To efficiently diversify the therapeutic uses of quercetin, Escherichia coli was harnessed as a production factory by the installation of various plant and bacterial UDP-xylose sugar biosynthetic genes. The genes encoding for the UDP-xylose pathway enzymes phosphoglucomutase (nfa44530), glucose-1-phosphate uridylyltransferase (galU), UDP-glucose dehydrogenase (calS8), and UDP-glucuronic acid decarboxylase (calS9) were overexpressed in E. coli BL21 (DE3) along with a glycosyltransferase (arGt-3) from Arabidopsis thaliana. Furthermore, E. coli BL21(DE3)/∆pgi, E. coli BL21(DE3)/∆zwf, E. coli BL21(DE3)/∆pgizwf, and E. coli BL21(DE3)/∆pgizwfushA mutants carrying the aforementioned UDP-xylose sugar biosynthetic genes and glycosyltransferase and the galU-integrated E. coli BL21(DE3)/∆pgi host harboring only calS8, calS9, and arGt-3 were constructed to enhance whole-cell bioconversion of exogeneously supplied quercetin into 3-O-xylosyl quercetin. Here, we report the highest production of 3-O-xylosyl quercetin with E. coli BL21 (DE3)/∆pgizwfushA carrying UDP-xylose sugar biosynthetic genes and glycosyltransferase. The maximum concentration of 3-O-xylosyl quercetin achieved was 23.78 mg/L (54.75 μM), representing 54.75 % bioconversion, which was an ~4.8-fold higher bioconversion than that shown by E. coli BL21 (DE3) with the same set of genes when the reaction was carried out in 5-mL culture tubes with 100 μM quercetin under optimized conditions. Bioconversion was further improved by 98 % when the reaction was scaled up in a 3-L fermentor at 36 h.

Keywords

Biotransformation E. coli Glycosylation Metabolic engineering Quercetin 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Ramesh Prasad Pandey
    • 1
  • Sailesh Malla
    • 2
    • 3
  • Dinesh Simkhada
    • 4
  • Byung-Gee Kim
    • 2
  • Jae Kyung Sohng
    • 1
  1. 1.Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical EngineeringSun Moon UniversityAsansiSouth Korea
  2. 2.Laboratory of Molecular Biotechnology and Biomaterials, School of Chemical and Biological EngineeringSeoul National UniversitySeoulSouth Korea
  3. 3.Novo Nordisk Foundation Center for BiosustainabilityTechnical University of DenmarkHørsholmDenmark
  4. 4.Department of ChemistryTexas A & M UniversityCollege StationUSA

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