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An enzymatic [4+2] cyclization cascade creates the pentacyclic core of pyrroindomycins

Nature Chemical Biology volume 11, pages 259–265 (2015)Cite this article

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Abstract

The [4+2] cycloaddition remains one of the most intriguing transformations in synthetic and natural products chemistry. In nature, however, there are remarkably few enzymes known to have this activity. We herein report an unprecedented enzymatic [4+2] cyclization cascade that has a central role in the biosynthesis of pyrroindomycins, which are pentacyclic spirotetramate natural products. Beginning with a linear intermediate that contains two pairs of 1,3-diene and alkene groups, the dedicated cyclases PyrE3 and PyrI4 act in tandem to catalyze the formation of two cyclohexene rings in the dialkyldecalin system and the tetramate spiro-conjugate of the molecules. The two cyclizations are completely enzyme dependent and proceed in a regio- and stereoselective manner to establish the enantiomerically pure pentacyclic core. Analysis of a related spirotetronate pathway confirms that homologs are functionally exchangeable, establishing the generality of these findings and explaining how nature creates diverse active molecules with similar rigid scaffolds.

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Figure 1: Spirotetramate and spirotetronate natural products and associated biosynthetic pathway.
Figure 2: Characterization of the dialkyldecalin synthase PyrE3 and the spiro-conjugate synthase PyrI4, which respectively represent two new types of cyclases catalyzing [4+2] cycloaddition reactions.
Figure 3: Validation of the functional identity of ChlE3 and ChlL to PyrE3 and PyrI4, respectively.
Figure 4: Known (putative) types of [4+2] cyclases and associated conversions in the pathways.

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Acknowledgements

This work was supported in part by grants from the National Natural Science Foundation of China (91213303, 81202453, 31430005 and 91413101), STCSM (14JC1407700 and 13XD1404500), the Ministry of Science and Technology of China (2012AA02A706) and the National Research Foundation (OTKA K105871) of Hungary.

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Author notes

  1. Zhenhua Tian, Peng Sun and Yan Yan: These authors contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China

    Zhenhua Tian, Peng Sun, Yan Yan, Zhuhua Wu, Qingfei Zheng, Shuaixiang Zhou, Hua Zhang, Futao Yu, Xinying Jia & Wen Liu

  2. Research Center for Marine Drugs, School of Pharmacy, Second Military Medical University, Shanghai, China

    Peng Sun

  3. Huzhou Center of Bio-Synthetic Innovation, Huzhou, China

    Dandan Chen & Wen Liu

  4. Department of Organic Chemistry, University of Debrecen, Debrecen, Hungary

    Attila Mándi & Tibor Kurtán

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  1. Zhenhua Tian
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Contributions

Z.T., Z.W., H.Z., X.J. and D.C. performed the in vivo investigations. Z.T., Y.Y., S.Z., F.Y. and Q.Z. conducted the in vitro enzymatic investigations. P.S., Y.Y. and Z.W. characterized the chemical compounds. A.M. and T.K. conducted the DFT conformational analysis and TDDFT-ECD calculation. Z.T. performed the sequence analysis. Z.T., P.S. and W.L. analyzed the data and wrote the manuscript. W.L. directed the research.

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Correspondence to Wen Liu.

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Supplementary Notes 1–3, Supplementary Figures 1–18 and Supplementary Tables 1–9. (PDF 6902 kb)

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Tian, Z., Sun, P., Yan, Y. et al. An enzymatic [4+2] cyclization cascade creates the pentacyclic core of pyrroindomycins. Nat Chem Biol 11, 259–265 (2015). https://doi.org/10.1038/nchembio.1769

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