Brief Communication | Published:

Monobactam formation in sulfazecin by a nonribosomal peptide synthetase thioesterase

Nature Chemical Biology volume 14, pages 57 (2018) | Download Citation

Abstract

The N-sulfonated monocyclic β-lactam ring characteristic of the monobactams confers resistance to zinc metallo-β-lactamases and affords the most effective class to combat carbapenem-resistant enterobacteria (CRE). Here we report unprecedented nonribosomal peptide synthetase activities, wherein an assembled tripeptide is N-sulfonated in trans before direct synthesis of the β-lactam ring in a noncanonical, cysteine-containing thioesterase domain. This means of azetidinone synthesis is distinct from the three others known in nature.

  • Compound

    N5-((R)-1-oxo-1-(((S)-2-oxoazetidin-3-yl)amino)propan-2-yl)-D-glutamine

  • Compound

    desmethoxylsulfazecin

  • Compound

    sulfazecin

  • Compound

    N5-((R)-1-(((S)-1-carboxy-2-(sulfoamino)ethyl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    N5-((R)-1-(((R)-3-hydroxy-2-oxo-1-sulfoazetidin-3-yl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    di-tert-butyl (R)-5-oxopyrrolidine-1,2-dicarboxylate

  • Compound

    (R)-5-(tert-butoxy)-4-((tert-butoxycarbonyl)amino)-5-oxopentanoic acid

  • Compound

    benzyl D-alaninate

  • Compound

    tert-butyl N5-((R)-1-(benzyloxy)-1-oxopropan-2-yl)-N2-(tert-butoxycarbonyl)-D-glutaminate

  • Compound

    (7S,10R,15R)-tert-butyl 7-((benzyloxy)carbonyl)-15-((tert-butoxycarbonyl)amino)-2,2,10-trimethyl-4,9,12-trioxo-3-oxa-5,8,11-triazahexadecan-16-oate

  • Compound

    tert-butyl N5-((2R)-1-(((7S,18R)-20-(((((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-4-hydroxy-3-(phosphonooxy)tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)-18-hydroxy-2,2,19,19-tetramethyl-4,8,13,17-tetraoxo-3-oxa-9-thia-5,12,16-triazaicosan-7-yl)amino)-1-oxopropan-2-yl)-N2-(tert-butoxycarbonyl)-D-glutaminate

  • Compound

    N5-((2R)-1-(((2S)-3-amino-1-((2-(3-((2R)-4-(((((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-4-hydroxy-3-(phosphonooxy)tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanamido)ethyl)thio)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    tert-butyl N5-((2R)-1-((2-(3-((2R)-4-(((((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-4-hydroxy-3-(phosphonooxy)tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanamido)ethyl)thio)-1-oxopropan-2-yl)-N2-(tert-butoxycarbonyl)-D-glutaminate

  • Compound

    N5-((2R)-1-((2-(3-((2R)-4-(((((((2R,3S,4R,5R)-5-(6-amino-9H-purin-9-yl)-4-hydroxy-3-(phosphonooxy)tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)oxy)(hydroxy)phosphoryl)oxy)-2-hydroxy-3,3-dimethylbutanamido)propanamido)ethyl)thio)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    benzyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-((tert-butoxycarbonyl)amino)propanoate

  • Compound

    benzyl (S)-2-amino-3-((tert-butoxycarbonyl)amino)propanoate

  • Compound

    tert-butyl N2-(tert-butoxycarbonyl)-N5-((R)-1-(((S)-2,2-dimethyl-4,8,13-trioxo-3-oxa-9-thia-5,12-diazatetradecan-7-yl)amino)-1-oxopropan-2-yl)-D-glutaminate

  • Compound

    N5-((R)-1-(((S)-1-((2-acetamidoethyl)thio)-3-amino-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    (R)-N-(3-((2-mercaptoethyl)amino)-3-oxopropyl)-2,2,5,5-tetramethyl-1,3-dioxane-4-carboxamide

  • Compound

    tert-butyl N2-(tert-butoxycarbonyl)-N5-((R)-1-(((7S,18R)-18,20-dihydroxy-2,2,19,19-tetramethyl-4,8,13,17-tetraoxo-3-oxa-9-thia-5,12,16-triazaicosan-7-yl)amino)-1-oxopropan-2-yl)-D-glutaminate

  • Compound

    N5-((R)-1-(((S)-3-amino-1-((2-(3-((R)-2,4-dihydroxy-3,3-dimethylbutanamido)propanamido)ethyl)thio)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    dibenzyl (R)-5-oxopyrrolidine-1,2-dicarboxylate

  • Compound

    (R)-5-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)-5-oxopentanoic acid

  • Compound

    benzyl N2-((benzyloxy)carbonyl)-N5-((R)-1-(tert-butoxy)-1-oxopropan-2-yl)-D-glutaminate

  • Compound

    tetrabutylammonium (S)-3-((R)-2-((R)-5-(benzyloxy)-4-(((benzyloxy)carbonyl)amino)-5-oxopentanamido)propanamido)-2-oxoazetidine-1-sulfonate

  • Compound

    benzyl (S)-(2-oxoazetidin-3-yl)carbamate

  • Compound

    tetrabutylammonium (S)-3-(((benzyloxy)carbonyl)amino)-2-oxoazetidine-1-sulfonate

  • Compound

    tetrabutylammonium (S)-3-amino-2-oxoazetidine-1-sulfonate

  • Compound

    benzyl (S)-3-((R)-2-((R)-5-(tert-butoxy)-4-((tert-butoxycarbonyl)amino)-5-oxopentanamido)propanamido)-2-oxoazetidine-1-carboxylate

  • Compound

    N5-((R)-1-(((S)-2-amino-1-carboxyethyl)amino)-1-oxopropan-2-yl)-D-glutamine

  • Compound

    tert-butyl (S)-(2-oxoazetidin-3-yl)carbamate

  • Compound

    benzyl (S)-3-((tert-butoxycarbonyl)amino)-2-oxoazetidine-1-carboxylate

  • Compound

    benzyl (S)-3-amino-2-oxoazetidine-1-carboxylate

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Primary accessions

NCBI Reference Sequence

References

  1. 1.

    , , , & Nat. Rev. Microbiol. 13, 42–51 (2015).

  2. 2.

    & Proc. Natl. Acad. Sci. USA 110, E653–E661 (2013).

  3. 3.

    , & Trends Mol. Med. 18, 263–272 (2012).

  4. 4.

    Curr. Opin. Chem. Biol. 35, 97–108 (2016).

  5. 5.

    , & Nat. Prod. Rep. 25, 757–793 (2008).

  6. 6.

    , & Nat. Prod. Rep. 29, 1074–1098 (2012).

  7. 7.

    Angew. Chem. Int. Edn Engl. 48, 4688–4716 (2009).

  8. 8.

    , & Nature 520, 383–387 (2015).

  9. 9.

    & Nat. Chem. Biol. 10, 251–258 (2014).

  10. 10.

    & Methods Enzymol. 458, 181–217 (2009).

  11. 11.

    et al. Nucleic Acids Res. 43, D222–D226 (2015).

  12. 12.

    , & Cell Chem. Biol. 24, 24–34 (2017).

  13. 13.

    et al. Chem. Biol. 21, 379–388 (2014).

  14. 14.

    & Biochem. J. 186, 613–616 (1980).

  15. 15.

    et al. Biochemistry 37, 1585–1595 (1998).

  16. 16.

    , & Science 284, 486–489 (1999).

  17. 17.

    Structure and Mechanism in Protein Science: A Guide to Enzyme Catalysis and Protein Folding (Freeman, New York, 1999).

  18. 18.

    et al. J. Biol. Chem. 286, 14445–14454 (2011).

  19. 19.

    et al. J. Am. Chem. Soc. 131, 16033–16035 (2009).

  20. 20.

    et al. Curr. Opin. Chem. Biol. 5, 525–534 (2001).

  21. 21.

    & Biochem. Soc. Trans. 44, 738–744 (2016).

  22. 22.

    , , , & Nature 521, 105–109 (2015).

  23. 23.

    , , & Curr. Opin. Struct. Biol. 41, 46–53 (2016).

  24. 24.

    J. Polym. Sci. A Polym. Chem. 44, 606–613 (2006).

  25. 25.

    Native-PAGE. in Biology Assays & Protocols Vol. 2017 (Sino Biological Inc., 2017).

Download references

Acknowledgements

This work was supported by the NIH (AI014937 and AI121072). We thank J. Liu of the University of North Carolina, Chapel Hill for providing DNA encoding PAPS synthesis proteins and I.P. Mortimer at the Johns Hopkins Mass Spectroscopy Facility for UPLC–HRMS data.

Author information

Affiliations

  1. Department of Chemistry, The Johns Hopkins University, Baltimore, Maryland, USA.

    • Ryan A Oliver
    • , Rongfeng Li
    •  & Craig A Townsend

Authors

  1. Search for Ryan A Oliver in:

  2. Search for Rongfeng Li in:

  3. Search for Craig A Townsend in:

Contributions

C.A.T. and R.A.O. designed and directed the study. R.A.O. carried out the syntheses and biochemical reactions. R.A.O. and R.F.L. cloned and expressed proteins. All authors analyzed the data and discussed the results. R.A.O., R.F.L., and C.A.T. prepared the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Craig A Townsend.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Results, Supplementary Table 1, Supplementary Figures 1–14

  2. 2.

    Life Sciences Reporting Summary

  3. 3.

    Supplementary Note

    General synthetic methods

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nchembio.2526