Abstract
We transformed an alkaloid biosynthetic gene with reengineered substrate specificity into Catharanthus roseus. The resulting transgenic plant cell culture produced a variety of unnatural alkaloid compounds when cocultured with simple, achiral, commercially available precursors that the reengineered enzyme was designed to accept. This work demonstrates the power of genetic engineering to retailor the structures of complex alkaloid natural products in plant culture.
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Acknowledgements
We gratefully acknowledge J. Shanks and C. Peebles (Iowa State) for detailed advice in the transformation procedure. CAMBIA is acknowledged for providing the pCAMBIA vectors, and N.-H. Chua (Rockefeller) is acknowledged for providing pTA7002. We thank L. Smeester (MIT) for assistance with rt-PCR, J.J. Maresh (MIT) for helpful discussions regarding the Agrobacterium transformation and N. Nims (MIT) for helpful suggestions regarding primer design for rt-PCR experiments. N. Yerkes (MIT) generously provided strictosidine standards. We thank T. Kutchan (Danforth Plant Science Center) for suggesting the pCAMBIA vector system. We gratefully acknowledge J. Simpson's (MIT) assistance in obtaining two-dimensional NMR data. This work was supported by the US National Science Foundation (MCB0719120). We acknowledge the US National Institutes of Health and the American Cancer Society for additional support.
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W.R. designed and performed all experiments and contributed to data analysis and manuscript writing. S.E.O. was the principal investigator of the project, contributed to data analysis and manuscript writing and provided funding.
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Supplementary Figures 1–7, Supplementary Table 1 and Supplementary Methods (PDF 1636 kb)
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Runguphan, W., O'Connor, S. Metabolic reprogramming of periwinkle plant culture. Nat Chem Biol 5, 151–153 (2009). https://doi.org/10.1038/nchembio.141
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DOI: https://doi.org/10.1038/nchembio.141
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