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Protein & Peptide Letters

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ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

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Research Article

Utilization of SUMO Tag and Freeze-thawing Method for a High-level Expression and Solubilization of Recombinant Human Angiotensinconverting Enzyme 2 (rhACE2) Protein in E. coli

Author(s): Mozafar Mohammadi*, Ramezan Ali Taheri, Peyman Bemani, Mohammad Sadegh Hashemzadeh, Gholamreza Farnoosh and Razieh Amini

Volume 29, Issue 7, 2022

Published on: 23 August, 2022

Page: [605 - 610] Pages: 6

DOI: 10.2174/0929866529666220715101357

Price: $65

Abstract

Background: SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) as a receptor for entering the host cells. Production of the ACE2 molecule is important because of its potency to use as a blocker and therapeutic agent against SARS-CoV-2 for the prophylaxis and treatment of COVID-19.

Objective: The recombinant human ACE2 (rhACE2) is prone to form an inclusion body when expressed in the bacterial cells.

Methods: We used the SUMO tag fused to the rhACE2 molecule to increase the expression level and solubility of the fusion protein. Afterward, the freeze-thawing method plus 2 M urea solubilized aggregated proteins. Subsequently, the affinity of solubilized rhACE2 to the receptor binding domain (RBD) of the SARS-CoV-2 spike was assayed by ELISA and SPR methods.

Results: SUMO protein succeeded in increasing the expression level but not solubilization of the fusion protein. The freeze-thawing method could solubilize and recover the aggregated fusion proteins significantly. Also, ELISA and SPR assays confirmed the interaction between solubilized rhACE2 and RBD with high affinity.

Conclusion: The SUMO tag and freeze-thawing method would be utilized for high-level expression and solubilization of recombinant rhACE2 protein.

Keywords: ACE2, inclusion body, mild solubilization, RBD, SARS-CoV-2, SUMO tag, rhACE2.

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