Discovery, characterization and engineering of ligases for amide synthesisCitation formats

  • External authors:
  • Michael Winn
  • Michael Rowlinson
  • Fanghua Wang
  • Luis Bering
  • Daniel Francis

Standard

Discovery, characterization and engineering of ligases for amide synthesis. / Winn, Michael; Rowlinson, Michael; Wang, Fanghua; Bering, Luis; Francis, Daniel; Levy, Colin; Micklefield, Jason.

In: Nature, Vol. 593, No. 7859, 19.05.2021, p. 391-398.

Research output: Contribution to journalArticlepeer-review

Harvard

Winn, M, Rowlinson, M, Wang, F, Bering, L, Francis, D, Levy, C & Micklefield, J 2021, 'Discovery, characterization and engineering of ligases for amide synthesis', Nature, vol. 593, no. 7859, pp. 391-398. https://doi.org/10.1038/s41586-021-03447-w

APA

Winn, M., Rowlinson, M., Wang, F., Bering, L., Francis, D., Levy, C., & Micklefield, J. (2021). Discovery, characterization and engineering of ligases for amide synthesis. Nature, 593(7859), 391-398. https://doi.org/10.1038/s41586-021-03447-w

Vancouver

Winn M, Rowlinson M, Wang F, Bering L, Francis D, Levy C et al. Discovery, characterization and engineering of ligases for amide synthesis. Nature. 2021 May 19;593(7859):391-398. https://doi.org/10.1038/s41586-021-03447-w

Author

Winn, Michael ; Rowlinson, Michael ; Wang, Fanghua ; Bering, Luis ; Francis, Daniel ; Levy, Colin ; Micklefield, Jason. / Discovery, characterization and engineering of ligases for amide synthesis. In: Nature. 2021 ; Vol. 593, No. 7859. pp. 391-398.

Bibtex

@article{bd2ab1658c014a6199f24af7981bd071,
title = "Discovery, characterization and engineering of ligases for amide synthesis",
abstract = "Coronatine and related bacterial phytotoxins are mimics of the hormone jasmonyl-L-isoleucine (JA-Ile), which mediates physiologically important plant signalling pathways1-4. Coronatine-like phytotoxins disrupt these essential pathways and have potential in the development of safer, more selective herbicides. Although the biosynthesis of coronatine has been investigated previously, the nature of the enzyme that catalyses the crucial coupling of coronafacic acid to amino acids remains unknown1,2. Here we characterize a family of enzymes, coronafacic acid ligases (CfaLs), and resolve their structures. We found that CfaL can also produce JA-Ile, despite low similarity with the Jar1 enzyme that is responsible for ligation of JA and L-Ile in plants5. This suggests that Jar1 and CfaL evolved independently to catalyse similar reactions-Jar1 producing a compound essential for plant development4,5, and the bacterial ligases producing analogues toxic to plants. We further demonstrate how CfaL enzymes can be used to synthesize a diverse array of amides, obviating the need for protecting groups. Highly selective kinetic resolutions of racemic donor or acceptor substrates were achieved, affording homochiral products. We also used structure-guided mutagenesis to engineer improved CfaL variants. Together, these results show that CfaLs can deliver a wide range of amides for agrochemical, pharmaceutical and other applications.",
author = "Michael Winn and Michael Rowlinson and Fanghua Wang and Luis Bering and Daniel Francis and Colin Levy and Jason Micklefield",
note = "Funding Information: Acknowledgements We thank the BBSRC (grants BB/K002341/1 and BB/N023536/1) and Syngenta for funding. F.W. was supported by the China Scholarship Council (grant no. 201806155100) and L.B. was funded by the Deutsche Forschungsgemeinschaft (DFG, grant BE 7054/1). The Michael Barber Centre for Collaborative Mass Spectrometry provided access to MS instrumentation. We also thank J. Vincent and N. Mulholland (Syngenta) for helpful discussions in the early stages of the project, and N. J. Turner (University of Manchester) for kindly providing the CHU plasmid. We also thank Diamond Light Source for beamtime access on i03 and i04-1 (proposal mx17773-56 and 76). Publisher Copyright: {\textcopyright} 2021, The Author(s), under exclusive licence to Springer Nature Limited.",
year = "2021",
month = may,
day = "19",
doi = "10.1038/s41586-021-03447-w",
language = "English",
volume = "593",
pages = "391--398",
journal = "Nature: international weekly journal of science",
issn = "0028-0836",
publisher = "Springer Nature",
number = "7859",

}

RIS

TY - JOUR

T1 - Discovery, characterization and engineering of ligases for amide synthesis

AU - Winn, Michael

AU - Rowlinson, Michael

AU - Wang, Fanghua

AU - Bering, Luis

AU - Francis, Daniel

AU - Levy, Colin

AU - Micklefield, Jason

N1 - Funding Information: Acknowledgements We thank the BBSRC (grants BB/K002341/1 and BB/N023536/1) and Syngenta for funding. F.W. was supported by the China Scholarship Council (grant no. 201806155100) and L.B. was funded by the Deutsche Forschungsgemeinschaft (DFG, grant BE 7054/1). The Michael Barber Centre for Collaborative Mass Spectrometry provided access to MS instrumentation. We also thank J. Vincent and N. Mulholland (Syngenta) for helpful discussions in the early stages of the project, and N. J. Turner (University of Manchester) for kindly providing the CHU plasmid. We also thank Diamond Light Source for beamtime access on i03 and i04-1 (proposal mx17773-56 and 76). Publisher Copyright: © 2021, The Author(s), under exclusive licence to Springer Nature Limited.

PY - 2021/5/19

Y1 - 2021/5/19

N2 - Coronatine and related bacterial phytotoxins are mimics of the hormone jasmonyl-L-isoleucine (JA-Ile), which mediates physiologically important plant signalling pathways1-4. Coronatine-like phytotoxins disrupt these essential pathways and have potential in the development of safer, more selective herbicides. Although the biosynthesis of coronatine has been investigated previously, the nature of the enzyme that catalyses the crucial coupling of coronafacic acid to amino acids remains unknown1,2. Here we characterize a family of enzymes, coronafacic acid ligases (CfaLs), and resolve their structures. We found that CfaL can also produce JA-Ile, despite low similarity with the Jar1 enzyme that is responsible for ligation of JA and L-Ile in plants5. This suggests that Jar1 and CfaL evolved independently to catalyse similar reactions-Jar1 producing a compound essential for plant development4,5, and the bacterial ligases producing analogues toxic to plants. We further demonstrate how CfaL enzymes can be used to synthesize a diverse array of amides, obviating the need for protecting groups. Highly selective kinetic resolutions of racemic donor or acceptor substrates were achieved, affording homochiral products. We also used structure-guided mutagenesis to engineer improved CfaL variants. Together, these results show that CfaLs can deliver a wide range of amides for agrochemical, pharmaceutical and other applications.

AB - Coronatine and related bacterial phytotoxins are mimics of the hormone jasmonyl-L-isoleucine (JA-Ile), which mediates physiologically important plant signalling pathways1-4. Coronatine-like phytotoxins disrupt these essential pathways and have potential in the development of safer, more selective herbicides. Although the biosynthesis of coronatine has been investigated previously, the nature of the enzyme that catalyses the crucial coupling of coronafacic acid to amino acids remains unknown1,2. Here we characterize a family of enzymes, coronafacic acid ligases (CfaLs), and resolve their structures. We found that CfaL can also produce JA-Ile, despite low similarity with the Jar1 enzyme that is responsible for ligation of JA and L-Ile in plants5. This suggests that Jar1 and CfaL evolved independently to catalyse similar reactions-Jar1 producing a compound essential for plant development4,5, and the bacterial ligases producing analogues toxic to plants. We further demonstrate how CfaL enzymes can be used to synthesize a diverse array of amides, obviating the need for protecting groups. Highly selective kinetic resolutions of racemic donor or acceptor substrates were achieved, affording homochiral products. We also used structure-guided mutagenesis to engineer improved CfaL variants. Together, these results show that CfaLs can deliver a wide range of amides for agrochemical, pharmaceutical and other applications.

U2 - 10.1038/s41586-021-03447-w

DO - 10.1038/s41586-021-03447-w

M3 - Article

C2 - 34012085

VL - 593

SP - 391

EP - 398

JO - Nature: international weekly journal of science

JF - Nature: international weekly journal of science

SN - 0028-0836

IS - 7859

ER -