Abstract
Polyketides are structurally diverse and medically important natural products that have various biological activities. During biosynthesis, chain elongation uses activated dicarboxylic acid building blocks, and their availability therefore limits side chain variation in polyketides. Recently, the crotonyl-CoA carboxylase-reductase (CCR) class of enzymes was identified in primary metabolism and was found to be involved in extender-unit biosynthesis of polyketides. These enzymes are, in theory, capable of forming dicarboxylic acids that show any side chain from the respective unsaturated fatty acid precursor. To our knowledge, we here report the first crystal structure of a CCR, the hexylmalonyl-CoA synthase from Streptomyces sp. JS360, in complex with its substrate. Structural analysis and biochemical characterization of the enzyme, including active site mutations, are reported. Our analysis reveals how primary metabolic CCRs can evolve to produce various dicarboxylic acid building blocks, setting the stage to use CCRs for the production of unique extender units and, consequently, altered polyketides.
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Acknowledgements
D.W.H. acknowledges support from the Fonds der Chemischen Industrie. Research in R.M.'s laboratory was funded by the Deutsche Forschungsgemeinschaft and the Bundesministerium für Bildung und Forschung (BMBF). We thank V. Wray for critically reviewing this manuscript. We also would like to thank T. Hoffmann for technical support, K. Harmrolfs for the NMR measurement and S.C. Wenzel for advice during the course of this project. In addition, we appreciate the work of A. Ullrich and K. Schultz from U. Kazmaier's work group for the syntheses of 2-octenoyl-CoA and 2-octenoyl-SNAC.
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N.Q. performed protein crystallization, crystallographic data collection and structure determination. L.H. performed gene cloning, protein heterologous expression and purification experiments as well as in vitro characterization of proteins and generation of corresponding point mutations. S.R. contributed initial advice regarding protein expression. R.M. and D.W.H. designed the study, and R.M. wrote the manuscript together with N.Q., L.H. and D.W.H. All authors discussed the results and commented on the manuscript.
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Quade, N., Huo, L., Rachid, S. et al. Unusual carbon fixation gives rise to diverse polyketide extender units. Nat Chem Biol 8, 117–124 (2012). https://doi.org/10.1038/nchembio.734
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DOI: https://doi.org/10.1038/nchembio.734
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