Figure 14.

Experiment-3 of the intestinal pancreatic trypsin (78 U/mL) effects (with describing model parameter estimates $\pm \mathrm{SE})$ on GH5-p4818Cel5_2A activity over a 5-h period after the intestinal pancreatic trypsin buffer being mixed with the GH5-p4818Cel5_2A enzyme 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-p4818Cel5_2A 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: $\mathrm{Y}=86.219\left(\pm ,3.969\right)-17.601\left(\pm ,6.123\right)\mathrm{x}+0.703(\pm 0.323)x^3$, P < 0.0001 for the intercept; P = 0.005 for the linear parameter estimate, P = 0.232 for the quadratic parameter estimate; and P = 0.032 for the cubic parameter estimate; ${\mathrm{R}}^2=0.883$, n = 84; and alternatively according to the linear response model established as $\mathrm{y}=78.177\left(\pm ,2.445\right)-15.559\left(\pm ,0.792\right)\mathrm{x}$, P < 0.0001 for all parameter estimates, ${\mathrm{r}}^2=0.830$, n = 84; (B) Plot of the inhibition kinetic relationship between the relative GH5-p4818Cel5_2A 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 ± 3.214) at 0-min exposure to the trypsin-78-U/mL incubation) and incubation time (t, h) under the trypsin (78 U/mL) condition established according to the cubic response model established as: $\mathrm{Y}=108.1\left(\pm ,4.977\right)-22.075\left(\pm ,7.679\right)\mathrm{x}+0.881(\pm 0.405)x^3$, P < 0.0001 for the intercept, P = 0.005 for the linear parameter; P = 0.232 for the quadratic parameter and P = 0.032 for the cubic parameter; ${\mathrm{R}}^2=0.883$, n = 84; and according to the linear response model established as $\mathrm{y}=98.044\left(\pm ,3.066\right)-19.513\left(\pm ,0.993\right)\mathrm{x}$, P < 0.0001 for all parameter estimates, ${\mathrm{r}}^2=0.830$, n = 84; (C) the Eadie-Hofstee linear plot between the GH5-p4818Cel5_2A enzyme activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{data}\mathrm{point},$ µmol/mg protein•min) and the v/t ratio established as: $\mathrm{y}=9.762\left(\pm ,1.158\right)+1.202\left(\pm ,0.047\right)\mathrm{x}$, P < 0.0001 for all parameter estimates; ${\mathrm{r}}^2=0.906$, n = 72; and (D) the Eadie-Hofstee linear plot between the relative GH5-p4818Cel5_2A enzyme activity (v, mean $\pm \mathrm{SE},\mathrm{n}=12\mathrm{at}\mathrm{each}\mathrm{data}\mathrm{point},$ % of the control group measured at I_c of 100.000 ± 3.214) at 0-min exposure to the trypsin-78-U/mL incubation) and the v/t ratio established as: $\mathrm{y}=12.244\left(\pm ,1.453\right)+1.202\left(\pm ,0.047\right)\mathrm{x}$, P < 0.0001 all parameter estimates, ${\mathrm{r}}^2=0.906$, n = 72.