## 3 Results
### Cationic Peptide Fractions Shows Multifunctional Bioactivity  
To assess the extent of its multifunctional ability, the cationic fraction was tested against ACE, DPP-IV, \(\alpha\)-amylase, and \(\alpha\)-glucosidase (Table 2). The DPP-IV inhibiting capacity was greater than 30% for all concentrations tested, never more than 50%, yet still exceeded the activity of the anionic fraction. The superior inhibiting activity was also observed for \(\alpha\)-amylase, \(\alpha\)-glucosidase, and ACE, with IC\({}_{50}\) values of 18.5, 13.9, and 1.9 \(\upmu\)g/mL, respectively. In contrast, the anionic fraction IC\({}_{50}\) values were 5082.7 \(\upmu\)g/mL (\(\alpha\)-amylase), 76.62 \(\upmu\)g/mL (\(\alpha\)-glucosidase), and 509 \(\upmu\)g/mL (ACE), indicating that the most potent peptides were present in the cationic fraction (Table 2). The data also illustrated that bioactive efficacy increased as fractionation progressed. Peptide fractions displayed enhanced \begin{table}
\begin{tabular}{c c c c c} \hline \hline  & **Fraction** & **ACE (\%)** & **DPP-IV (\%)** & \(\alpha\)**-Glucosidase (\%)** \\ \hline \multirow{4}{*}{**Size exclusion**} & S1 & \(32.3\pm 0.31\)\({}^{\text{a}}\) & \(19.3\pm 2.52\)\({}^{\text{a}}\) & \(62.2\pm 1.09\)\({}^{\text{a}}\) \\ \cline{2-5}  & S2 & \(28.8\pm 0.02\)\({}^{\text{b}}\) & \(16.98\pm 0.30\)\({}^{\text{a}}\) & \(25.9\pm 2.54\)\({}^{\text{b}}\) \\ \cline{2-5}  & S3 & \(47.5\pm 0.87\)\({}^{\text{c}}\) & \(20.6\pm 0.61\)\({}^{\text{a}}\) & \(83.6\pm 1.14\)\({}^{\text{c}}\) \\ \cline{2-5}  & S4 & \(73.9\pm 1.12\)\({}^{\text{d}}\) & \(68.0\pm 2.52\)\({}^{\text{b}}\) & \(89.5\pm 0.91\)\({}^{\text{d}}\) \\ \hline \multirow{4}{*}{**Reverse-phase**} & F1 & \(25.4\pm 0.6\)\({}^{\text{e}}\) & \(5.1\pm 0.01\)\({}^{\text{c}}\) & \(9.4\pm 0.36\)\({}^{\text{e}}\) \\ \cline{2-5}  & F2 & \(29.4\pm 0.04\)\({}^{\text{g}}\) & \(15.3\pm 2.70\)\({}^{\text{d}}\) & \(11.6\pm 0.04\)\({}^{\text{f}}\) \\ \cline{2-5}  & F3 & \(67.2\pm 0.12\)\({}^{\text{h}}\) & \(18.4\pm 0.59\)\({}^{\text{a}}\) & \(8.4\pm 0.47\)\({}^{\text{g}}\) \\ \cline{2-5}  & F4 & \(32.2\pm 0.19\)\({}^{\text{a}}\) & \(17.1\pm 1.61\)\({}^{\text{a}}\) & \(7.3\pm 1.51\)\({}^{\text{h}}\) \\ \cline{2-5}  & F5 & \(47.4\pm 0.5\)\({}^{\text{c}}\) & \(21.0\pm 1.96\)\({}^{\text{a}}\) & \(22.3\pm 2.79\)\({}^{\text{i}}\) \\ \cline{2-5}  & F6 & \(93.4\pm 0.08\)\({}^{\text{i}}\) & \(26.98\pm 1.63\)\({}^{\text{e}}\) & \(48.2\pm 5.79\)\({}^{\text{j}}\) \\ \hline \multirow{4}{*}{**Positive inhibitor**} & Captopril & \(98.0\pm 0.01\)\({}^{\text{j}}\) & n.a & n.a \\  & Ile-Leu-Pro & n.a & \(88.9\pm 0.05\)\({}^{\text{f}}\) & n.a \\ \cline{1-1}  & Acarbose & n.a & n.a & \(70.9\pm 0.43\)\({}^{\text{k}}\) \\ \hline \hline \end{tabular}  
* Results are reported as the percentage of inhibiting activity and is expressed as a mean of least triplicate (n = 3) determinations, with standard deviation. S1-S4 are fractions collected from the gel filtration column. F1–F6 are fractions collected from the reverse phase C18 column. Captopril, peptide Ile-Leu-Pro, and Acarbose are commercially available therapeutic options to inhibit the angiotensin-converting enzyme II, Dipeptidyl peptidase-4, and \(\alpha\)-glucosidase, respectively. n.a. = not applicable. CPHD = CPH after simulated digestion with gastrointestinal enzymes. Lowercase letters indicate significant differences between fractions within assays (\(p<0.05\)).  
\end{table} 
Table 1: Percentage inhibition of angiotensin converting enzyme (ACE), dipeptidyl peptidase-IV (DPP-IV), and \(\alpha\)-glucosidase by CPH-digests (CPHD) peptide fractions. inhibiting ability after being separated into charged groups (anion and cation), as compared to peptide fractions after SEC and C18 separation. This was true for all enzymes tested, except DPP-IV. Treating RAW264.7 macrophage cells with LPS resulted in a significant increase (\(p<0.05\)) in inflammatory induction, compared to that in the non-treated control.