Bombyx batryticatus Cocoonase Inhibitor Separation, Purification, and Inhibitory Effect on the Proliferation of SMCC-7721 HeLa-Derived Cells

Figure 2

Detection and kinetic analysis of the inhibitory activity of BBCI on silkworm cocoonase. (a) Detection of inhibitory activity. Following the addition of 1, 2, 3, 4, 5, 6, or 7 nM BBCI to the reaction system containing 5 nM (120 μg) cocoonase and 1 μM substrate BAPNA, respectively, A₄₁₀ was measured. Taking the content of BBCI as the abscissa, and the average value of the remaining activity of cocoonase (A_BBCI/A_control × 100%) of the three repeated experiments as the ordinate, the graph was drawn using Origin 2018 software. (b) Kinetic analysis. Following the addition of 20, 40, 60, 80, 100, 120, 140, 160, or 180 nM BAPNA to the reaction system containing 5 nM (120 μg) cocoonase and 2 or 3 nM BBCI, respectively, A₄₁₀ was measured. Taking the substrate BAPNA concentrations as the abscissa, and the average of the reaction speed V (V = 2.58 × 10⁻⁵A₄₁₀) of three repeated experiments as the ordinate, the Michaelis–Menten model was used for mapping using Origin 2018 software. A₄₁₀: light absorption value at 410 nm; BBCI: Bombyx batryticatus cocoonase inhibitor.

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