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High level production and characterization of truncated human angiotensin converting enzyme 2 in Nicotiana benthamiana plant as a potential therapeutic target in COVID-19

Tarlan Mamedov, İrem Gürbüzaslan, Merve Ilgin, Damla Yuksel, Gunay Mammadova, Aykut Ozkul, Gulnara Hasanova
doi: https://doi.org/10.1101/2021.05.17.444533
Tarlan Mamedov
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
2Azerbaijan National Academy of Science, Department of Biology and Medical Science, 24 Istiglaliyyat Street, Baku, Azerbaijan
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  • For correspondence: tmammedov@gmail.com
İrem Gürbüzaslan
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
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Merve Ilgin
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
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Damla Yuksel
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
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Gunay Mammadova
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
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Aykut Ozkul
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
3Department of Virology, Faculty of Veterinary Medicine, Ankara University, Ankara 06110, Turkey
4Biotechnology Institute, Ankara University, Ankara 06135, Turkey
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Gulnara Hasanova
1Akdeniz University, Department of Agricultural Biotechnology, Dumlupınar Boulevard 07058 Campus, Antalya, Turkey
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Abstract

The COVID-19 pandemic, which is caused by SARS-CoV-2 has rapidly spread to more than 222 countries and has put global public health at high risk. The world urgently needs safe, a cost-effective SARS-CoV-2 coronavirus vaccine, therapeutic and antiviral drugs to combat the COVID-19. Angiotensin-converting enzyme 2 (ACE2), as a key receptor for SARS-CoV-2 infections, has been proposed as a potential therapeutic target in COVID-19 patients. In this study, we report high level production (about ∼0.75 g /kg leaf biomass) of glycosylated and non-glycosylated forms of recombinant human truncated ACE2 in Nicotiana benthamiana plant. The plant produced recombinant human truncated ACE2s successfully bind to the SARC-CoV-2 spike protein, but deglycosylated ACE2 binds more strongly than the glycosylated counterpart. Importantly, both deglycosylated and glycosylated forms of AEC2 stable at elevated temperatures for prolonged periods and demonstrated strong anti-SARS-CoV-2 activity in vitro. The IC50 values of glycosylated and deglycosylated AEC2 were 0.4 and 24 μg/ml, respectively, for the pre-entry infection, when incubated with 100TCID50 of SARS-CoV-2. Thus, plant produced truncated ACE2s are promising cost-effective and safe candidate as a potential therapeutic targets in the treatment of COVID-19 patients.

Competing Interest Statement

T. M. named inventor on patent applications covering plant produced ACE2. All other authors have no any competing financial and/or non-financial interests.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 17, 2021.
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High level production and characterization of truncated human angiotensin converting enzyme 2 in Nicotiana benthamiana plant as a potential therapeutic target in COVID-19
Tarlan Mamedov, İrem Gürbüzaslan, Merve Ilgin, Damla Yuksel, Gunay Mammadova, Aykut Ozkul, Gulnara Hasanova
bioRxiv 2021.05.17.444533; doi: https://doi.org/10.1101/2021.05.17.444533
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High level production and characterization of truncated human angiotensin converting enzyme 2 in Nicotiana benthamiana plant as a potential therapeutic target in COVID-19
Tarlan Mamedov, İrem Gürbüzaslan, Merve Ilgin, Damla Yuksel, Gunay Mammadova, Aykut Ozkul, Gulnara Hasanova
bioRxiv 2021.05.17.444533; doi: https://doi.org/10.1101/2021.05.17.444533

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