Figure 12.

Experiment-3 of the intestinal pancreatic trypsin (78 U/mL) effects (with describing model parameter estimates $\pm \mathrm{SE})$ on GH5-tCel5A1 activity over a 5-h period after the intestinal pancreatic trypsin buffer being mixed with the GH5-tCel5A1enzyme preparation for incubation at the resulting mixture pH = 6.0–7.0 and trypsin (78 U/mL) prior to further incubation for measuring cellulase activity with the carboxymethyl cellulose (CMC) substrate buffer in presence of dithiothreitol (DTT) (5 mM) and N₂ gas purging for 15 min at 37 °C. (A) Plot of the inhibition kinetic relationship between the GH5-tCel5A1 activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{time}\mathrm{point},$ nmol/mg protein•min) and incubation time (t, h) under the trypsin (78 U/mL) condition established according to the cubic response model as: $\mathrm{Y}=16.140\left(\pm ,2.166\right)-13.659\left(\pm ,3.342\right)\mathrm{x}+4.087\left(\pm ,1.451\right)x^2-0.378(\pm 0.176)x^3$, P < 0.0001 for the intercept; P = 0.0001 for the linear parameter; P = 0.006 for the quadratic parameter estimate; and P = 0.035 for the cubic parameter, ${\mathrm{R}}^2=0.326$, n = 84; and alternatively, according to the linear regression model as: $\mathrm{y}=7.930\left(\pm ,1.251\right)-1.459\left(\pm ,0.405\right)\mathrm{x}$, P < 0.0001 for the intercept; and P = 0.001 for the slope, ${\mathrm{r}}^2=0.142$, n = 84; (B) Plot of the inhibition kinetic relationship between the relative GH5-tCel5A1 activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{time}\mathrm{point},$ % of the control group measured at I_c of 100.000 ± 18.294 at 0-min exposure to the trypsin-78-U/mL incubation) and incubation time (t, h) established according to the cubic response model: $\mathrm{Y}=122.6\left(\pm ,16.458\right)-103.8\left(\pm ,25.393\right)\mathrm{x}+31.050\left(\pm ,11.026\right)x^2-2.875(\pm 1.338)x^3$, P < 0.0001 for the intercept; P = 0.0001 for the linear parameter estimate; P = 0.006 for the quadratic parameter estimate; and P = 0.035 for the cubic parameter estimate, ${\mathrm{R}}^2=0.326$, n = 84; and alternatively, according to the linear regression model established as: $\mathrm{y}=60.251\left(\pm ,9.5\right)-11.091\left(\pm ,3.077\right)\mathrm{x}$, P < 0.0001 for the intercept; and P = 0.001 for the slope; ${\mathrm{r}}^2=0.142$, n = 84; (C) the Eadie-Hofstee linear plot between the GH5-tCel5A1 enzyme activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{data}\mathrm{point},$ nmol/mg protein•min) and the v/t ratio established as: $\mathrm{y}=0.681\left(\pm ,0.251\right)+1.639\left(\pm ,0.090\right)\mathrm{x}$, P = 0.008 for the intercept; and P < 0.0001 for the slope, ${\mathrm{r}}^2=0.829$, n = 72; and (D) the Eadie-Hofstee linear plot between the relative GH5-tCel5A1 enzyme activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{data}\mathrm{point},$ % of the control group measured at Ic of 100.000 ± 18.294) at 0-min exposure to the trypsin-78-U/mL incubation) incubation] and the v/t ratio established as $\mathrm{y}=5.173\left(\pm ,1.904\right)+1.639\left(\pm ,0.090\right)\mathrm{x}$, P = 0.008 for the intercept; and P < 0.0001 for the slope; ${\mathrm{r}}^2=0.829$, n = 72.