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Production of 3-O-xylosyl quercetin in Escherichia coli

  • Biotechnological products and process engineering
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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)/∆pgi∆zwf, and E. coli BL21(DE3)/∆pgi∆zwf∆ushA 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)/∆pgi∆zwf∆ushA 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.

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Acknowledgment

This work was supported a grant from the Next-Generation BioGreen 21 Program (SSAC, grant no. PJ008013), Rural Development Administration and the Converging Research Center Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (20090082333), Republic of Korea.

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Authors and Affiliations

  1. Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University, #100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam, 336-708, South Korea

    Ramesh Prasad Pandey & Jae Kyung Sohng

  2. Laboratory of Molecular Biotechnology and Biomaterials, School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea

    Sailesh Malla & Byung-Gee Kim

  3. Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark

    Sailesh Malla

  4. Department of Chemistry, Texas A & M University, College Station, TX, 77843, USA

    Dinesh Simkhada

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  1. Ramesh Prasad Pandey
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  2. Sailesh Malla
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  3. Dinesh Simkhada
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  4. Byung-Gee Kim
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  5. Jae Kyung Sohng
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Correspondence to Jae Kyung Sohng.

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Pandey, R.P., Malla, S., Simkhada, D. et al. Production of 3-O-xylosyl quercetin in Escherichia coli . Appl Microbiol Biotechnol 97, 1889–1901 (2013). https://doi.org/10.1007/s00253-012-4438-9

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  • DOI: https://doi.org/10.1007/s00253-012-4438-9

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