earticle

영어

Acute respiratory syndrome coronavirus SARS-CoV-2 causing COVID-19 has been rapidly spreading worldwide since it occurred in December 2019. SARS-CoV-2 infection occurs as the receptor-binding domain (RBD) of the coronavirus envelope spike protein binds to the angiotensin-converting enzyme 2 (ACE2) receptor. ACE2 is a cell membrane protein activated when it is cleaved and released from the cell surface. The soluble ACE2 can act as a decoy receptor to neutralize the SARS-CoV-2 virus, which is a potential therapy for COVID-19. In this study, the recombinant ACE2 protein was produced in a transient Nicotiana benthamiana plant expression system. The ACE2 was cloned to a fragment crystallizable (Fc) tagged with the KDEL sequence, endoplasmic reticulum (ER) retention signal (ACE2-FcK) in pEAQ-HT, the Cowpea mosaic virus (CPMV)-based transient plant expression vector. To design the recombinant protein structure of ACE2-FcK, the transmembrane region of ACE2 was removed using a transmembrane region prediction program. Four N-glycosylation sites recognized were identified using the N-glycosylation prediction program. Finally, Agrobacterium (LBA4404) carrying pEAQ-HT ACE2-FcK vector was applied for transient expression of ACE2-FcK in N. benthamiana. The proteins (100kDa) were successfully expressed and purified from leaves using protein A affinity chromatography. Additionally, suggested that the plant-derived ACE2-FcK has bioactivities as a therapeutic agent for coronavirus. Taken together, the ACE2-FcK protein can be transiently produced with an optimized expression time and leaf position post-infiltration in plant.