Efficient in planta production of amidated antimicrobial peptides that are active against drug-resistant ESKAPE pathogens

Shahid Chaudhary, Zahir Ali, Muhammad Tehseen, Evan F. Haney, Aarón Pantoja-Angles, Salwa Alshehri, Tiannyu Wang, Gerard J. Clancy, Maya Ayach, Charlotte Hauser, Pei Ying Hong, Samir M. Hamdan, Robert E.W. Hancock, Magdy Mahfouz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Antimicrobial peptides (AMPs) are promising next-generation antibiotics that can be used to combat drug-resistant pathogens. However, the high cost involved in AMP synthesis and their short plasma half-life render their clinical translation a challenge. To address these shortcomings, we report efficient production of bioactive amidated AMPs by transient expression of glycine-extended AMPs in Nicotiana benthamiana line expressing the mammalian enzyme peptidylglycine α-amidating mono-oxygenase (PAM). Cationic AMPs accumulate to substantial levels in PAM transgenic plants compare to nontransgenic N. benthamiana. Moreover, AMPs purified from plants exhibit robust killing activity against six highly virulent and antibiotic resistant ESKAPE pathogens, prevent their biofilm formation, analogous to their synthetic counterparts and synergize with antibiotics. We also perform a base case techno-economic analysis of our platform, demonstrating the potential economic advantages and scalability for industrial use. Taken together, our experimental data and techno-economic analysis demonstrate the potential use of plant chassis for large-scale production of clinical-grade AMPs.

Original languageEnglish (US)
Article number1464
JournalNature Communications
Volume14
Issue number1
DOIs
StatePublished - Dec 2023

Bibliographical note

Funding Information:
This work was supported by BAS/1/1035-01-01 baseline and KAUST Smart Health Initiative (KSHI) funding to MM with additional support to REWH from the Canadian Institutes for Health Research FDN-154287 to REWH. We would like to thank Prof. Betty Eipper at University of Connecticut Health Center, USA for the generous gift of plasmid DNA encoding rat variants of PAM enzymes. We would like to thank Dr. Roger Plaut from Center of Biologics and Research, US Food and Drug Administration, for the generous gift of luminescent version of clinical S. aureus USA300 strain NRS384. We would like to thank Prof. Frank Sainsbury and Prof. George Lomonossoff for the generous gift of pEAQ-HT vector. We would like to thank Prof. Dirk Görlich for the generous gift of plasmid expressing SENP protease (pAV0679, Addgene plasmid #149689). We would also like to thank Miriam Amah, Senior Technical Specialist of KAUST, for helping with ESI-MS optimization. EuH

Funding Information:
This work was supported by BAS/1/1035-01-01 baseline and KAUST Smart Health Initiative (KSHI) funding to MM with additional support to REWH from the Canadian Institutes for Health Research FDN-154287 to REWH. We would like to thank Prof. Betty Eipper at University of Connecticut Health Center, USA for the generous gift of plasmid DNA encoding rat variants of PAM enzymes. We would like to thank Dr. Roger Plaut from Center of Biologics and Research, US Food and Drug Administration, for the generous gift of luminescent version of clinical S. aureus USA300 strain NRS384. We would like to thank Prof. Frank Sainsbury and Prof. George Lomonossoff for the generous gift of pEAQ-HT vector. We would like to thank Prof. Dirk Görlich for the generous gift of plasmid expressing SENPEuHprotease (pAV0679, Addgene plasmid #149689). We would also like to thank Miriam Amah, Senior Technical Specialist of KAUST, for helping with ESI-MS optimization.

Publisher Copyright:
© 2023, The Author(s).

ASJC Scopus subject areas

  • General Chemistry
  • General Biochemistry, Genetics and Molecular Biology
  • General Physics and Astronomy

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