Abstract
Consumption of cruciferous vegetables is associated with reduced risk of developing cancer, a phenomenon attributed to glucosinolates, which are characteristic of these vegetables. We report production of the bioactive benzylglucosinolate in the noncruciferous plant Nicotiana benthamiana through metabolic engineering. The study includes identification of γ-glutamyl peptidase 1 (GGP1), which substantially increased glucosinolate production by metabolizing an accumulating glutathione conjugate, an activity not previously described for glucosinolate biosynthesis or for proteins containing glutamine amidotransferase domains.
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Acknowledgements
We thank the Danish International Developmental Agency for a PhD stipend to F.G.-F. (DANIDA project no. 91175) and the Villum Kann Rasmussen Fond for its support to the VKR Research Centre for Pro-Active Plants. We also thank Novozymes for the Novo Scholarship to M.T.N. and M.E.M.
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F.G.-F. and B.A.H. formulated the project; M.T.N. and M.E.M. performed experiments in N. benthamiana; F.G.-F. and M.N. expressed GGP1 in E. coli and performed kinetic measurements; M.S.M. synthesized GS-B; C.E.O. performed LC-MS analysis; F.G.-F. and M.T.N. analyzed the data; F.G.-F., M.T.N. and B.A.H. wrote the manuscript.
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The heterologous production of glucosinolates is the subject of a PCT patent application filed 27 February 2009 (PCT/IB2009/000500).
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Supplementary Figures 1–3, Supplementary Scheme 1 and Supplementary Methods (PDF 309 kb)
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Geu-Flores, F., Nielsen, M., Nafisi, M. et al. Glucosinolate engineering identifies a γ-glutamyl peptidase. Nat Chem Biol 5, 575–577 (2009). https://doi.org/10.1038/nchembio.185
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DOI: https://doi.org/10.1038/nchembio.185
