# Studying the Process of Enzyme Treatment on Beef Meat-Bone Paste Quality
## 3 Results and Discussion
### Physicochemical Analysis  
The chemical composition of meat-bone paste before and after enzyme treatment showed no significant differences (Table 1). The enzymatic action, in conjunction with other factors like the influence of ascorbic acid, can facilitate the loosening of collagen structures and enhance the hydration of proteins. Pepsin specifically targets collagen, a major protein component of bone. Breaking down collagen into smaller peptides could contribute to higher protein values in the analysis [48]. Additionally, the enzymatic treatment appears to induce changes in the fat component of the meat-bone paste. The splitting of fat cells during treatment may contribute to the observed slight decrease in fat content in MBP-E compared to MBP-C, although the difference is minimal. The slight decrease in ash content after enzymatic treatment might be due to solubilization and leaching of some minerals by the enzymes and acids. The pH of the MBP-E sample was lower than that of the MBP-C sample, indicating a more acidic environment due to the enzyme treatment process. This decrease in pH could be attributed to the addition of ascorbic acid during enzyme treatment, which can influence the functional properties of the meat-bone paste (Table 2). \begin{table}
\begin{tabular}{c c c} \hline \hline
**Indicator in Percent** & **MBP-C** & **MBP-E** \\ \hline Protein \% & 13.70 \(\pm\) 0.28 & 14.12 \(\pm\) 0.23 \\ \hline Fat \% & 4.35 \(\pm\) 0.05 & 4.23 \(\pm\) 0.06 \\ \hline Ash \% & 15.98 \(\pm\) 0.19 & 15.47 \(\pm\) 0.28 \\ \hline Water \% & 65.97 \(\pm\) 0.85 & 66.18 \(\pm\) 1.19 \\ \hline MBP-C—meat-bone paste without enzyme treatment (control); MBP-E—meat-bone paste after enzyme treatment. \\ \end{tabular}
\end{table} 
Table 1: Chemical composition of meat-bone paste. \begin{table}
\begin{tabular}{c c c} \hline \hline
**Indicator** & **MBP-C** & **MBP-E** \\ \hline pH & 7.02 \(\pm\) 0.12 b  & 5.9 \(\pm\) 0.09 a  \\ \hline Water-binding capacity (\%) & 77.28 \(\pm\) 1.50 a  & 82.54 \(\pm\) 1.05 b  \\ \hline Viscosity (Pa \(\times\) s) & 13.35 \(\pm\) 0.16 b  & 11.27 \(\pm\) 0.13 a  \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Physical properties of meat-bone paste. Additionally, the water-binding capacity of MBP-E was higher than that of the control sample, suggesting that the enzyme treatment process led to an increased ability of the meat-bone paste to retain water. This could be due to the breakdown of protein structures during enzyme action, resulting in improved water-binding capacity. This is explained by the fact that in the treatment process, the proteins are hydrated, and the amount of immobilized moisture in the meat-bone paste increases due to the loosening of collagen structure and splitting of fat cells. The loosening of collagen promotes additional binding of water molecules [49]. [PERSON] et al. (2024) also noted an increase in the water-binding capacity of protein hydrolysates after enzymatic treatment with microbial culture of chicken gizzards [50]. The decrease in viscosity of MBP-E compared to the control sample indicates that the enzyme action process altered the rheological properties of the meat-bone paste, possibly due to the changes in protein and water interactions caused by the enzymatic activity during enzyme treatment. The decline in viscosity observed in MBP-E can be attributed to two primary factors. Firstly, the increase in water-binding capacity in MBP-E contributes to a more hydrated matrix, leading to a reduction in the viscosity of the meat-bone paste. The enhanced water-binding capacity indicates that the enzyme action process promotes the incorporation of moisture into the paste, altering its rheological behavior. Secondly, the decrease in viscosity is linked to the structural changes induced by the enzymatic action of pepsin. The enzymatic action of pepsin, in conjunction with ascorbic acid, leads to destructive changes in collagen fibers within the meat-bone paste.