Abstract
We have characterized four sequential enzymes that transform 1-hydroxy-N-methylcanadine to narcotoline hemiacetal, completing our elucidation of noscapine biosynthesis in opium poppy. Two cytochromes P450 catalyze hydroxylations at C13 and C8 on the protoberberine scaffold, the latter step inducing ring opening and the formation of an aldehyde moiety. Acetylation at C13 before C8 hydroxylation introduces a protective group subsequently hydrolyzed by a carboxylesterase, which triggers rearrangement to a cyclic hemiacetal.
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References
Lettré, H. Ann. NY Acad. Sci. 58, 1264–1275 (1954).
Ye, K. et al. Proc. Natl. Acad. Sci. USA 95, 1601–1606 (1998).
Mahmoudian, M. & Rahimi-Moghaddam, P. Recent Pat. Anticancer Drug Discov. 4, 92–97 (2009).
Fang, Z.Z. et al. Br. J. Pharmacol. 167, 1271–1286 (2012).
Battersby, A.R. & Hirst, M. Tetrahedr. Lett. 11, 669–673 (1965).
Battersby, B.A.R., Staunton, J. & Wiltshire, H.R. J. Chem. Soc. Perkin Trans. I 1975, 1147–1156 (1975).
Dang, T.-T.T. & Facchini, P.J. Plant Physiol. 159, 618–631 (2012).
Winzer, T. et al. Science 336, 1704–1708 (2012).
Dang, T.-T.T. & Facchini, P.J. FEBS Lett. 588, 198–204 (2014).
Dang, T.-T.T. & Facchini, P.J. J. Biol. Chem. 289, 2013–2026 (2014).
Chen, X. & Facchini, P.J. Plant J. 77, 173–184 (2014).
Dinesh-Kumar, S.P., Anandalakshmi, R., Marathe, R., Schiff, M. & Liu, Y. Methods Mol. Biol. 236, 287–294 (2003).
D'Auria, J.C. Curr. Opin. Plant Biol. 9, 331–340 (2006).
Gershater, M.C. & Edwards, R. Plant Sci. 173, 579–588 (2007).
Ruppert, M. et al. Planta 222, 888–898 (2005).
Grothe, T., Lenz, R. & Kutchan, T.M. J. Biol. Chem. 276, 30717–30723 (2001).
Onoyovwe, A. et al. Plant Cell 25, 4110–4122 (2013).
McGary, K.L. et al. Proc. Natl. Acad. Sci. USA 110, 11481–11486 (2013).
Shulgin, A.T. & Perry, W.E. The Simple Plant Isoquinolines (Transform Press, Berkeley, CA, 2003).
Nguyen, D.T. et al. J. Biol. Chem. 285, 16588–16598 (2010).
Ro, D.-K. et al. BMC Biotechnol. 8, 83 (2008).
Dang, T.-T.T., Onoyovwe, A., Farrow, S.C. & Facchini, P.J. Methods Enzymol. 515, 231–266 (2012).
Hileman, L.C., Drea, S., Martino, G., Litt, A. & Irish, V.F. Plant J. 44, 334–341 (2005).
Acknowledgements
We thank D. Burns and J. Hagel for the NMR and LTQ-Orbitrap analyses, respectively. T.-T.T.D. is the recipient of the Alberta Innovates Technology Futures and University of Calgary Eyes High graduate scholarships. Funding was provided through grants from Genome Canada, Genome Alberta, the Government of Alberta (URSI-10-004-SEG), the Canada Foundation for Innovation (CFI-21695) and the Natural Sciences and Engineering Research Council of Canada (RGPIN/183573-2009) to P.J.F.
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T.-T.T.D. and X.C. contributed equally to all experiments. P.J.F. was responsible for experimental design. All authors contributed to the preparation of the manuscript.
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P.J.F., T.-T.T.D. and X.C. have filed a patent application (Patent Cooperation Treaty Application PCT/CA2014/050782) covering compositions and methods related to the manufacture of noscapine and synthesis intermediates thereof.
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Supplementary Text and Figures
Supplementary Results, Supplementary Figures 1–18 and Supplementary Tables 1–7. (PDF 20145 kb)
Supplementary Data Set 1
1H, 13C, COSY, HSQC, HMBC, and NOESY NMR spectral data for (13S,14R)-1,13-dihydroxy-N-methylcanadine (m/z 386) and 1H, COSY, HSQC and HMBC NMR spectral data for (13S,14R)-1-hydroxy-13-O-acetyl-N-methylcanadine (m/z 428). (PDF 898 kb)
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Dang, TT., Chen, X. & Facchini, P. Acetylation serves as a protective group in noscapine biosynthesis in opium poppy. Nat Chem Biol 11, 104–106 (2015). https://doi.org/10.1038/nchembio.1717
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DOI: https://doi.org/10.1038/nchembio.1717
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