Fig. 3
Characterization of rGAPDH: determination of Km and Vmax for G3P and NAD+ of a, b MT-GAPDH-H and c, d MS-GAPDH-H. The extent of enzymatic conversion was monitored by the formation of NADH at 340 nm at 25 °C. a, c The substrate NAD+ was added in excess (1 mM) while G3P was varied (0.02–2 mM). b, d Substrate G3P was in excess (2 mM), while NAD+ was varied from 0.01 to 2 mM. Data is fitted to Michaelis–Menten equation. Inset Data represented as a Lineweaver–Burk plot. e Comparative CD spectra of rGAPDH purified of MT-GAPDH-H and EC-H-GAPDH. f The activity was measured for 2 min, data is plotted as % residual activity versus pH. Average of three independent experiments is represented. g Temperature stability of rGAPDH-His. Data is plotted as % residual activity representing the average of three independent experiments. h Sensitivity of MT-GAPDH-H and M.tb H37Rv cytosol to H2O2 in the presence or absence of β-ME. i Sensitivity of MT-GAPDH-H to metal ions in the presence or absence of β-ME. Data is represented as % residual activity ±SD, n = 3