# Antimicrobial and Antioxidant Properties of Bovine Livers and Hearts Hydrolysates
## 3 Results and Discussion
### Amino Acids
An increase in FRAP activity with increasing hydrolysis time was also recorded in various protein hydrolyses obtained from meat by-products [11]. [PERSON] et al. found that using the papain enzyme and red blood cells as the substrate, FRAP activity increased in proportion to the duration of hydrolysis [35]. \begin{table}
\begin{tabular}{c c c c c c} \hline \multicolumn{2}{c}{**Bovine Liver**} & \multicolumn{4}{c}{**Bovine Heart**} \\ \hline
**Amino Acids** & **Time (h)** & **Pepsin** & **Papain** & **Pepsin** & **Papain** \\ \hline \multirow{4}{*}{Tyr} & 3 & 0.09 \(\pm\) 0.04 & 0.03 \(\pm\) 0.01 & 0.08 \(\pm\) 0.03 & 0.05 \(\pm\) 0.01 \\  & 6 & 0.04 \(\pm\) 0.03 & 0.02 \(\pm\) 0.01 & 0.07 \(\pm\) 0.01 & 0.03 \(\pm\) 0.01 \\  & 24 & 0.01 \(\pm\) 0.01 & 0.02 \(\pm\) 0.01 & 0.02 \(\pm\) 0.03 & 0.02 \(\pm\) 0.01 \\  & \(p\)-_value (Time)_ & * & & & * \\ \hline \multirow{4}{*}{Phe} & 3 & 0.13 \(\pm\) 0.02 & 0.11 \(\pm\) 0.01 & 0.12 \(\pm\) 0.03 & 0.15 \(\pm\) 0.04 \\  & 6 & 0.11 \(\pm\) 0.04 & 0.10 \(\pm\) 0.09 & 0.08 \(\pm\) 0.01 & 0.13 \(\pm\) 0.03 \\  & 24 & 0.09 \(\pm\) 0.04 & 0.02 \(\pm\) 0.01 & 0.05 \(\pm\) 0.01 & 0.02 \(\pm\) 0.01 \\  & \(p\)-_value (Time)_ & * & & * & * \\ \hline \multirow{4}{*}{His} & 3 & 0.14 \(\pm\) 0.02 & 0.11 \(\pm\) 0.03 & 0.11 \(\pm\) 0.02 & 0.17 \(\pm\) 0.03 \\  & 6 & 0.11 \(\pm\) 0.01 & 0.09 \(\pm\) 0.01 & 0.08 \(\pm\) 0.01 & 0.16 \(\pm\) 0.02 \\  & 24 & 0.07 \(\pm\) 0.01 & 0.05 \(\pm\) 0.02 & 0.05 \(\pm\) 0.01 & 0.05 \(\pm\) 0.01 \\  & \(p\)-_value (Time)_ & * & * & * & * \\ \hline \multirow{4}{*}{Lys} & 3 & 0.32 \(\pm\) 0.14 & 0.34 \(\pm\) 0.13 & 0.38 \(\pm\) 0.06 & 0.42 \(\pm\) 0.15 \\  & 6 & 0.22 \(\pm\) 0.13 & 0.27 \(\pm\) 0.04 & 0.33 \(\pm\) 0.08 & 0.38 \(\pm\) 0.11 \\  & 24 & 0.19 \(\pm\) 0.05 & 0.15 \(\pm\) 0.04 & 0.18 \(\pm\) 0.02 & 0.27 \(\pm\) 0.02 \\  & \(p\)-_value (Time)_ & * & * & * & * \\ \hline \multirow{4}{*}{Arg} & 3 & 0.28 \(\pm\) 0.02 & 0.25 \(\pm\) 0.04 & 0.31 \(\pm\) 0.05 & 0.31 \(\pm\) 0.02 \\  & 6 & 0.22 \(\pm\) 0.01 & 0.20 \(\pm\) 0.05 & 0.25 \(\pm\) 0.08 & 0.28 \(\pm\) 0.05 \\  & 24 & 0.21 \(\pm\) 0.06 & 0.19 \(\pm\) 0.03 & 0.19 \(\pm\) 0.03 & 0.23 \(\pm\) 0.03 \\  & \(p\)-_value (Time)_ & & * & * & * \\ \hline \end{tabular}
\end{table} 
Table 1: _Cont._Peptides make up the majority of hydrolysates, which is what gives them their antioxidant capacity because they can also contain endogenous substances like ascorbic acid, which could boost the total antioxidant capacity [32]. Furthermore, the antioxidant activity seen in the hydrolysates can be attributed to alterations in the structural proteins that occur as a result of enzymatic hydrolysis. This phenomenon can be attributed to the capacity of amino acids to engage in interactions with oxidizing agents, resulting in modifications and subsequent consequences for the residues [36]. Data from previous studies show that more antioxidant peptides are formed when the hydrolysis time is longer, and this has significantly higher DPPH scavenging activity [32]. On the other hand, the results of our study show that as the hydrolysis time increases, the amounts of all tested free amino acids tend to decrease, and they are inversely correlated with the antioxidant activity of hydrolysates of bovine hearts and livers, regardless of the enzyme used, but the strongest inverse correlation was established in a case of bovine hearts hydrolysates with papain (r = 0.856, \(p\) < 0.001). Antibacterial activity was determined using the agar diffusion assay (inhibition zone assay) (Table 3). The bovine heart hydrolysates exhibited the most significant antibacterial effects against _L. monocytogenes_ (20.0 \(\pm\) 0.2 mm), with considerable inhibition observed against _E. coli_ (10.0 \(\pm\) 0.1 mm) and _S. enterica_ subsp. _etteria_ serovar Typhimurium (10.0 \(\pm\) 0.1 mm) after 24 h of hydrolysis with papain. Bovine livers hydrolysates with papain 24 h. also exhibited stronger antibacterial efficacy against _L. monocytogenes_ (17.5 \(\pm\) 0.2 mm), _S. enterica_ subsp. _etteria_ serovar Typhimurium_ (10.0 \(\pm\) 0.1 mm).