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. 2022 Oct 3;3(4):101794. doi: 10.1016/j.xpro.2022.101794

Figure 2.

Figure 2

Production of active SARS-CoV-2 main protease (Mpro) in E. coli

(A) Bacterial expression and purification of Mpro. The SDS-PAGE gel was stained with coomassie brilliant blue R250. M: Protein ladder; 1: Total cell extract; 2–4: Pure Mpro (34 kDa).

(B) Measurement of the initial velocity of Mpro enzyme reaction. All initial rates were obtained by fitting the linear portion of the curves to a straight line.

(C) Determination of the Km value. The Michaelis-Menten equation was plotted using GraphPad Prism 8.0 according to the initial rates, and the Km and Vmax values were recorded as 19.28 μM and 139.5 ΔRFU/s, respectively. This assay was performed in triplicate.