Expression strategies for the efficient synthesis of antimicrobial peptides in plastids

Matthijs P. Hoelscher, Joachim Forner, Silvia Calderone, Carolin Krämer, Zachary Taylor, F. Vanessa Loiacono, Shreya Agrawal, Daniel Karcher, Fabio Moratti, Xenia Kroop and Ralph Bock ()
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Matthijs P. Hoelscher: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Joachim Forner: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Silvia Calderone: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Carolin Krämer: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Zachary Taylor: Max-Planck-Institut für Molekulare Pflanzenphysiologie
F. Vanessa Loiacono: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Shreya Agrawal: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Daniel Karcher: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Fabio Moratti: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Xenia Kroop: Max-Planck-Institut für Molekulare Pflanzenphysiologie
Ralph Bock: Max-Planck-Institut für Molekulare Pflanzenphysiologie

Nature Communications, 2022, vol. 13, issue 1, 1-17

Abstract: Abstract Antimicrobial peptides (AMPs) kill microbes or inhibit their growth and are promising next-generation antibiotics. Harnessing their full potential as antimicrobial agents will require methods for cost-effective large-scale production and purification. Here, we explore the possibility to exploit the high protein synthesis capacity of the chloroplast to produce AMPs in plants. Generating a large series of 29 sets of transplastomic tobacco plants expressing nine different AMPs as fusion proteins, we show that high-level constitutive AMP expression results in deleterious plant phenotypes. However, by utilizing inducible expression and fusions to the cleavable carrier protein SUMO, the cytotoxic effects of AMPs and fused AMPs are alleviated and plants with wild-type-like phenotypes are obtained. Importantly, purified AMP fusion proteins display antimicrobial activity independently of proteolytic removal of the carrier. Our work provides expression strategies for the synthesis of toxic polypeptides in chloroplasts, and establishes transplastomic plants as efficient production platform for antimicrobial peptides.

Date: 2022
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DOI: 10.1038/s41467-022-33516-1

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