Abstract
The SARS-CoV-2 coronavirus expresses two essential proteases: firstly, the 3Chymotrypsin-like protease (3CLpro) or main protease (Mpro), and secondly, the papain-like protease (PLpro), both of which are considered as viable drug targets for the inhibition of viral replication. In order to perform drug discovery assays for SARS-CoV-2, it is imperative that efficient methods are established for the production and purification of 3CLpro and PLpro of SARS-CoV-2, designated as 3CLpro-CoV2 and PLpro-CoV2, respectively. This article expands the data collected in the attempts to express SARS-CoV-2 proteases under different conditions and purify them under single-step chromatography. Data showed that the use of E. coli BL21(DE3) strain was sufficient to express 3CLpro-CoV2 in a fully soluble form. Nevertheless, the single affinity chromatography step was only applicable for 3CLpro-CoV2 expressed at 18 °C, with a yield and purification fold of 92% and 49, respectively. Meanwhile, PLpro-CoV2 was successfully expressed in a fully soluble form in either BL21(DE3) or BL21-CodonPlus(DE3) strains. In contrast, the single affinity chromatography step was only applicable for PLpro-CoV2 expressed using E. coli BL21-CodonPlus(DE3) at 18 or 37 °C, with a yield and purification fold of 86% (18 °C) or 83.36% (37 °C) and 112 (18 °C) or 71 (37 °C), respectively. The findings provide a guide for optimizing the production of SARS-CoV-2 proteases of E. coli host cells.
Highlights
Data showed that the use of E. coli BL21(DE3) strain was sufficient to express 3Chymotrypsinlike protease (3CLpro)-CoV2 in a fully soluble form
The single affinity chromatography step was only applicable for papain-like protease (PLpro)-CoV2 expressed using E. coli BL21-CodonPlus(DE3) at 18 or 37 ◦ C, with a yield and purification fold of 86% (18 ◦ C) or 83.36% (37 ◦ C) and 112 (18 ◦ C) or 71 (37 ◦ C), respectively
The findings provide a guide for optimizing the production of SARS-CoV-2 proteases of E. coli host cells
Summary
Since its first emergence in December 2019, a new coronavirus, namely, severe acute respiratory syndrome–coronavirus 2 (SARS-CoV-2), has become a global health issue [1,2,3]. The production of target protein through recombinant technology is often challenged by the issues of the expression level and purification process [15]. The whole process to obtain high purity of target protein under recombinant technology involves multiple and lengthy steps This includes, but is not limited to, gene cloning, transformation into the host cells, expression induction, host cells harvesting and lysis, removal of the cell debris, and end up with purification through one or more chromatography processes followed by the purity and yield determination [16]. This paper, offers a comprehensive description of the data of protein produced obtained from multiple expression conditions of SARS-CoV-2 proteases (Table 1). The first step involved the over-expression of 3CLpro-CoV2 and PLpro-CoV2 in the E. coli host cells under several conditions.
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