## 3 Results and Discussion
### Purification and Characterization of \(\alpha\)AI-1
\begin{table}
\begin{tabular}{c c c c c c c c c c c} \hline \hline
**Huunita Dry Bean** & \multicolumn{2}{c}{**Bush Bean**} & \multicolumn{2}{c}{**Red Kidney Bean**} & \multicolumn{2}{c}{**Pinto Bean**} & \multicolumn{2}{c}{**Pinto Bean**} & \multicolumn{2}{c}{**Pinto Bean**} \\
**“Purified \(\alpha\)AI-1** & **“Purified \(\alpha\)AI-1** & **“Purified \(\alpha\)AI-1** & **“Purified \(\alpha\)AI-1** & **“Purified \(\alpha\)AI-1** & **“Pruified \(\alpha\)AI-1** & **“Real** & **“Cooled Meal** \\  & **Protein”** & **Protein”** & **Protein”** & **Protein”** & **Protein”** & **Protein”** & **Protein”** & **Protein Bean** & **Protein Bean** & **Protein Bean** \\ \hline \(\mathrm{t_{R}}\) **(s)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** & **t (\%)** \\ \hline
24.97 & 3.52 & 24.91 & 8.34 & 25.04 & 1.41 & 24.92 & 8.28 & 24.10 & 1.65 & 24.04 & 0.96 \\
26.74 & 28.30 & 26.13 & 39.92 & 26.85 & 25.40 & 26.22 & 40.66 & 24.40 & 1.42 & 24.37 & 0.91 \\
27.40 & 10.20 & 26.69 & 20.40 & 27.43 & 13.98 & 26.79 & 20.25 & 25.22 & 2.12 & 25.22 & 1.49 \\
28.35 & 50.51 & 27.46 & 8.62 & 28.31 & 49.93 & 27.52 & 8.07 & 25.81 & 2.76 & 25.79 & 2.22 \\
30.07 & 3.97 & 27.81 & 9.37 & 30.12 & 2.99 & 27.93 & 9.55 & 26.25 & 3.95 & 26.22 & 5.75 \\  & & 28.48 & 3.56 & & & 28.51 & 3.56 & 26.59 & 2.65 & 27.07 & 2.68 \\  & & 29.19 & 1.15 & & & 29.23 & 1.20 & 27.03 & 2.28 & 27.36 & 2.90 \\  & & 30.01 & 6.68 & & & 30.12 & 6.84 & 27.22 & 2.61 & 27.74 & 1.34 \\  & & & & & & & & 27.71 & 3.94 & 28.31 & 4.43 \\  & & & & & & & & 28.39 & 4.78 & 29.11 & 3.91 \\  & & & & & & & & 29.12 & 4.43 & 30.20 & 18.46 \\  & & & & & & & & 30.10 & 16.01 & & \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Retention times (\(\mathrm{t_{R}}\)) and relative amounts of proteins in total protein (t, %) related to \(\alpha\)AI-1 peaks of the bean-derived “purified \(\alpha\)AI-1 protein” and bean meals detected by LOC electrophoresis. The Effect of In Vitro Gastric Digestion on the Presence, Immune Reactivity, and Bioactivity of \(\alpha\)AI-1  
The structural configuration of \(\alpha\)-AI significantly influences its functionality [3]. The Pinto-bean-derived \"purified \(\alpha\)AI-1 protein\" sample was submitted under in vitro digestion process (duration time: 2, 10, 30, and 60 min) using simulated gastric fluid (SGF). The resistance of \(\alpha\)AI-1 to the pepsin and acidic circumstances was investigated and monitored by SDS-PAGE (Figure 2). BSA (positive control) and \(\beta\)-lactoglobulin (BLG, negative control) was also submitted under digestion with the same conditions in order to verify the adequacy of the digestion process. BSA had already degraded after 2 min (as no any protein bands were detectable in the BSA molecular weight range (\(\sim\)66.4 kDa)). Figure 1: Protein pattern of LOC electrophoresis of _”purified \(\alpha\)AI-1 protein”_ samples of common beans. **Lanes:** L. molecular marker; 1. Huanita Dry bean \"purified \(\alpha\)AI-1 protein\"; 2. Bush bean \"purified \(\alpha\)AI-1 protein\"; 3. Red Kidney bean \"purified \(\alpha\)AI-1 protein\"; 4. Pinto bean \"purified \(\alpha\)AI-1 protein\". Figure 2: Monitoring the effect of gastric in vitro digestion (for 2, 10, 30, and 60 min) on the resistance of the Pinto-bean-derived “purified \(\alpha\)AI-1 protein” to digestion by 6/15% SDS-PAGE. **Lanes:** 1. pepsin; 2. Pinto bean \"purified \(\alpha\)AI-1 protein\"; 3. BSA; 4. \(\beta\)LG; 5. Pinto bean \"purified \(\alpha\)AI-1 protein\" after 2-min digestion; 6. BSA after 2-min digestion; 7. \(\beta\)LG after 2-min digestion; 8. Pinto bean \"purified \(\alpha\)AI-1 protein\" after 10-min digestion; 9. BSA after 10-min digestion; 10. \(\beta\)LG after 10-min digestion; 11. Pinto bean \"purified \(\alpha\)AI-1 protein\" after 30-min digestion; 12. BSA after 30-min digestion; 13. Pinto bean \"purified \(\alpha\)AI-1 protein\" after 60-min digestion; 15. BSA after 60-min digestion; 16. \(\beta\)LG after 60-min digestion.