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A two-step sulfation in antibiotic biosynthesis requires a type III polyketide synthase

Nature Chemical Biology volume 9, pages 610–615 (2013)Cite this article

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Abstract

Caprazamycins (CPZs) belong to a group of liponucleoside antibiotics inhibiting the bacterial MraY translocase, an essential enzyme involved in peptidoglycan biosynthesis. We have recently identified analogs that are decorated with a sulfate group at the 2″-hydroxy of the aminoribosyl moiety, and we now report an unprecedented two-step sulfation mechanism during the biosynthesis of CPZs. A type III polyketide synthase (PKS) known as Cpz6 is used in the biosynthesis of a group of new triketide pyrones that are subsequently sulfated by an unusual 3′-phosphoadenosine-5′-phosphosulfate (PAPS)-dependent sulfotransferase (Cpz8) to yield phenolic sulfate esters, which serve as sulfate donors for a PAPS-independent arylsulfate sulfotransferase (Cpz4) to generate sulfated CPZs. This finding is to our knowledge the first demonstration of genuine sulfate donors for an arylsulfate sulfotransferase and the first report of a type III PKS to generate a chemical reagent in bacterial sulfate metabolism.

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Figure 1: Structures of CPZs and sulfated CPZs, and genetic organization of the CPZ biosynthetic gene cluster (cpz9-cpz31).
Figure 2: HPLC profiles of S. coelicolor M512 mutant extracts.
Figure 3: Characterization of Cpz8 as PAPS-dependent sulfotransferase.
Figure 4: HPLC profiles of S. coelicolor M145/Δsco7221 mutant extracts.
Figure 5: Identification and structures of presulficidins.
Figure 6: In vitro analysis of the CPZ two step sulfation mechanism.

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Acknowledgements

The authors thank G. Challis (University of Warwick) for providing S. coelicolor M145/Δsco7221 and germicidin A. We are grateful to R. Machinek and C. Zolke (Institute of Organic Chemistry, University of Göttingen) for carrying out NMR measurements. We also thank A. Jones for reviewing the manuscript. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (SFB766) to K.E. and X.T., a grant from the Graduate School 'Promotionsverbund Antibakterielle Wirkstoffe' of the University of Tuebingen to X.T. and by the European Commission (IP005224, ActinoGen) to L.K.

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Authors and Affiliations

  1. Pharmaceutical Institute, University of Tübingen, Tübingen, Germany

    Xiaoyu Tang, Kornelia Eitel, Leonard Kaysser & Bertolt Gust

  2. Institute of Microbiology and Infection Medicine, University of Tübingen, Tübingen, Germany

    Andreas Kulik

  3. Institute of Organic Chemistry, University of Tübingen, Tübingen, Germany

    Stephanie Grond

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  1. Xiaoyu Tang
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Contributions

X.T., L.K. and B.G. designed the research. X.T. and L.K. generated and analyzed the mutants. X.T. performed the biochemical experiments and purified presulficidins. X.T. and K.E. purified hydroxyacylcaprazol and purified the proteins. X.T., K.E., L.K. and B.G. analyzed the data. A.K. performed MS analysis. S.G. elucidated the structure of presulficidins. X.T., S.G. and B.G. wrote the manuscript. B.G. supervised the project.

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Correspondence to Bertolt Gust.

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Supplementary Results, Supplementary Figures 1–15, Supplementary Note and Supplementary Tables 1–3. (PDF 2162 kb)

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Tang, X., Eitel, K., Kaysser, L. et al. A two-step sulfation in antibiotic biosynthesis requires a type III polyketide synthase. Nat Chem Biol 9, 610–615 (2013). https://doi.org/10.1038/nchembio.1310

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