Abstract
Objectives
To produce flavonol and flavone 6-C-glucosides by bioconversion using recombinant Escherichia coli expressing a C-glucosyltransferase from wasabi (WjGT1).
Results
Escherichia coli expressing WjGT1 (Ec-WjGT1) converted flavones (apigenin and luteolin) and flavonols (quercetin and kaempferol) into their 6-C-glucosides in M9 minimal media supplemented with glucose, and released these products into the culture media. Ec-WjGT1 system also converts a flavanone (naringenin) into its C-glucoside at a conversion rate of 60% in 6 h. For scale-up production, apigenin, kaempferol, and quercetin were sequentially fed into the Ec-WjGT1 system at concentrations of 20–50 µM every 15–60 min, and the system was then able to produce isovitexin, kaempferol 6-C-glucoside, and quercetin 6-C-glucoside at an 89–99% conversion rate.
Conclusions
The Ec-WjGT1 system quickly and easily produces flavone and flavonol 6-C-glucosides at high conversion rates when using sequential administration to avoid precipitation of substrates.
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Acknowledgements
We are indebted to the Division of Gene Research, Research Center for Advanced Science and Technology, Shinshu University, and the Research Center for Advanced Plant Factory, Shinshu University, for supplying the necessary research facilities. We also thank the Global leader program for fiber renaissance, Shinshu University for essential support to NJ, and Dr. Makoto Shimosaka, Shinshu University for a critical evaluation of the manuscript.
Supplementary Information
Supplementary Table 1—NMR assignment of the purified compounds produced by bioconversion (400 MHz, dimethyl sulfoxide-d6 (DMSO-d6)).
Supplementary Figure 1—Bioconversion of flavones by the Ec-WjGT1 system.
Supplementary Figure 2—Effect of glucose concentration on the bioconversion of apigenin by the Ec-WjGT1 system.
Supplementary Figure 3—Heteronuclear multiple bond correlation (HMBC) analysis of the purified compounds produced by bioconversion.
Supplementary Figure 4—Bioconversion of resveratrol by the Ec-WjGT1 system.
Funding
This work was supported in part by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers JP26450120 and JP20K05825 to GT.
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Dorjjugder, N., Hatano, M. & Taguchi, G. Production of flavonol and flavone 6-C-glucosides by bioconversion in Escherichia coli expressing a C-glucosyltransferase from wasabi (Eutrema japonicum). Biotechnol Lett 43, 1913–1919 (2021). https://doi.org/10.1007/s10529-021-03165-3
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DOI: https://doi.org/10.1007/s10529-021-03165-3