Iterative Mechanism of Macrodiolide Formation in the Anticancer Compound Conglobatin
Murphy, Annabel C
Dias, Luiz Carlos
Chemistry & Biology
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Zhou, Y., Murphy, A. C., Samborskyy, M., Prediger, P., Dias, L. C., & Leadlay, P. (2015). Iterative Mechanism of Macrodiolide Formation in the Anticancer Compound Conglobatin. Chemistry & Biology, 22 745-754. https://doi.org/10.1016/j.chembiol.2015.05.010
Conglobatin is an unusual C2-symmetrical macrodiolide from the bacterium Streptomyces conglobatus with promising antitumor activity. Insights into the genes and enzymes that govern both the assembly-line production of the conglobatin polyketide and its dimerization are essential to allow rational alterations to be made to the conglobatin structure. We have used a rapid, direct in vitro cloning method to obtain the entire cluster on a 41-kbp fragment, encoding a modular polyketide synthase assembly line. The cloned cluster directs conglobatin biosynthesis in a heterologous host strain. Using a model substrate to mimic the conglobatin monomer, we also show that the conglobatin cyclase/thioesterase acts iteratively, ligating two monomers head-to-tail then re-binding the dimer product and cyclizing it. Incubation of two different monomers with the cyclase produces hybrid dimers and trimers, providing the first evidence that conglobatin analogs may in future become accessible through engineering of the polyketide synthase.
We gratefully acknowledge BBSRC (project grant BB/J007250/1 to P.F.L.), the European Commission (Marie Curie Fellowship to Y.Z.), and the University of Cambridge (Herchel Smith Research Fellowship to A.C.M.), and Ms. Asha Boodhun (Department of Chemistry, University of Cambridge) for help in HR-MS analysis. L.C.D. acknowledges the support of Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, Proc. 2012/04616-3 and 2012/02230-0). P.F.L. is an International Research Awardee of the Alexander von Humboldt Foundation.
External DOI: https://doi.org/10.1016/j.chembiol.2015.05.010
This record's URL: https://www.repository.cam.ac.uk/handle/1810/248576
Attribution 2.0 UK: England & Wales
Licence URL: http://creativecommons.org/licenses/by/2.0/uk/
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