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. 2022 Jun 1;12:9135. doi: 10.1038/s41598-022-13124-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Experiment-2 of the gastric acidic pH effects (pH = 3.5) (with describing model parameter estimates $\pm SE)$ on GH5-tCel5A1 activity over a 5-h period after the gastric pH buffer being mixed with the crude GH5-tCel5A1 enzyme preparation 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) Exponential plot of the inhibition kinetic relationship between GH5-tCel5A1 activity (v, mean $\pm SE, n = 12 at each time point,$ nmol/mg protein•min) and incubation time (t, h) under gastric pH 3.5 established conditions according to the exponential response model as: $Y = 48.398 (\pm 5.710) e^{[- 0.099 (\pm 0.049) x)]}$ , P < 0.0001 for the initial residual tCel5A1 activity estimate; and P = 0.045 for the rate constant estimate; $R^{2} = 0.080$ , n = 84; and alternatively, according to the linear response model established as $y = 48.644 (\pm 5.127) - 4.193 (\pm 1.782) x$ , P < 0.0001 for the intercept; and P = 0.021 for the slope, $r^{2} = 0.087$ , n = 84; (B) Exponential plot of the inhibition kinetic relationship between relative GH5-tCel5A1 activity (v, mean $\pm SE, n = 12 at each time point,$ % of the control group measured (100.000 ± 19.72) at 0-min exposure to the gastric pH-3.5 incubation) and incubation time (t, h) under gastric pH 3.5 conditions according to the exponential response model established as: $Y = 107.3 (\pm 12.664) e^{[- 0.099 (\pm 0.049) x)]}$ , P < 0.0001 for the initial residual GH5-tCel5A1 activity estimate; and P = 0.045 for the rate constant estimate; $R^{2} = 0.080$ , n = 84; and according to the linear response model established as $y = 107.9 (\pm 11.369) - 9.299 (\pm 3.952) x$ , P < 0.0001 for all parameter estimates, $r^{2} = 0.087$ , n = 84; (C) the Eadie-Hofstee linear plot between the GH5-tCel5A1 enzyme activity [v, mean $\pm SE, n = 12 at each data point,$ nmol/mg protein•min] and the v/t ratio established as $y = 17.637 (\pm 3.067) - 1.008 (\pm 0.113) x$ , P < 0.0001 for all parameter estimates; $r^{2} = 0.564$ , n = 72; and (D) the Eadie-Hofstee linear plot between the relative GH5-tCel5A1 enzyme activity [v, mean $\pm SE, n = 12 at each data point,$ % of the control group, I_c at 100.000 ± 19.72) measured at 0-min exposure gastric pH-3.5 incubation] and the v/t ratio established as $y = 39.114 (\pm 6.801) - 1.008 (\pm 0.113) x$ , P < 0.0001 for all parameter estimates;, $r^{2} = 0.564$ , n = 72.