Optimization of Engineered Production of the Glucoraphanin Precursor Dihomomethionine in Nicotiana benthamiana

Frontiers in Bioengineering and Biotechnology
Christoph CrocollBarbara Ann Halkier

Abstract

Glucosinolates are natural products characteristic of the Brassicales order, which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Toward our goal of creating a rich source of glucoraphanin for dietary supplements, we have previously reported the feasibility of engineering glucoraphanin in Nicotiana benthamiana through transient expression of glucoraphanin biosynthetic genes from A. thaliana (Mikkelsen et al., 2010). As side-products, we obtained fivefold to eightfold higher levels of chain-elongated leucine-derived glucosinolates, not found in the native plant. Here, we investigated two different strategies to improve engineering of the methionine chain elongation part of the glucoraphanin pathway in N. benthamiana: (1) coexpression of the large subunit (LSU1) of the heterodimeric isopropylmalate isomerase and (2) coexpression of BAT5 transporter for efficient transfer of intermediates across the chloroplast membrane. We succeeded in raising dihomomethionine (DHM) levels to a maximum of 432 nmol g(-1) fresh weight that is equivalent to a ninefold increase compared to the h...Continue Reading

References

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Mar 23, 2010·Trends in Plant Science·Ida E SønderbyBarbara A Halkier
May 12, 2010·Molecular Plant·Michael Dalgaard MikkelsenBarbara Ann Halkier
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Sep 28, 2015·Metabolic Engineering·Nadia MirzaBarbara Ann Halkier

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Citations

Aug 7, 2018·Journal of Integrative Plant Biology·Annette PetersenBarbara Ann Halkier
Sep 4, 2018·The Plant Journal : for Cell and Molecular Biology·Dawei MaC Peter Constabel
May 23, 2018·Plant Cell Reports·James Reed, Anne Osbourn
Jun 27, 2019·Cold Spring Harbor Perspectives in Biology·Thomas Louveau, Anne Osbourn
Jul 19, 2017·Chembiochem : a European Journal of Chemical Biology·Johan Andersen-RanbergRubini Kannangara
Oct 28, 2020·Nature Chemical Biology·Ricardo De La Peña, Elizabeth S Sattely
Aug 15, 2021·Proceedings of the National Academy of Sciences of the United States of America·Jianbei HuangHenrik Hartmann

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Methods Mentioned

BETA
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