# Enhanced Enzymatic Degradability of Livestock Blood Pretreated with Ultrasonic Technique
###### Abstract  
Livestock blood, a major organic waste generated by the Livestock industry, poses a risk of pollution due to its rapid decomposition. However, it is a potential protein source for agricultural purposes due to its protein-rich organic matter. In this study, we conducted studies on an eco-friendly, scalable, and effective protein degradation process using Livestock blood to reduce waste and produce an amino acid Liquid fertilizer that can be recycled for agricultural use. Ultrasonic technology was applied as a pretreatment method to improve the enzymatic hydrolysis efficiency of proteins in Livestock blood, and the optimal conditions that Led to 95.91% solubilization rate of hemoglobin were ultrasound duration for 30 min at an ultrasound density of 0.5 W/mL. As a result of hydrolyzing ultrasonically pretreated blood by mixing exo- and endo-type proteolytic enzymes, the optimal combination was a mixture of Savinase\({}^{\text{\text{\textregistered}}}\) 1% and Flavourzyme\({}^{\text{\textregistered}}\) 1%. After 4 h of reaction, the protein concentration was 27.8 mg/mL and the amino acid concentration was confirmed to be 54.6 mg/mL. This is about 4.2 times higher than the amino acid concentration of blood without ultrasound pretreatment, 13.1 mg/mL, and it was confirmed that sonication has a significant effect on improving protein degradation efficiency. As protein degradation increased, the viscosity of blood gradually decreased, suggesting that the physical force applied to the agitator torque diminished during the enzyme reaction; a significant correlation between protein and amino acid concentrations (biological factors) and torque (mechanical factor) was observed. Measuring torque during an enzyme reaction can confirm the extent of the enzyme reaction, so it can be used as an indicator of reaction progress when scaling up the process in the future. livestock blood; ultrasonic solubilization; enzymatic degradation; amino acid Liquid fertilizer +
Footnote †: journal: Medical  
antimicrobial properties [7; 8; 9]. Therefore, the hydrolysis of Livestock blood to produce high-quality amino acids and eco-friendly fertilizer contributes to waste reduction in the Livestock industry and the transition to environmentally friendly agriculture. High-quality amino acids can be produced under milder conditions through the utilization of proteolytic enzymes to create amino acid Liquid fertilizer from Livestock blood, which is more efficient than the chemical decomposition methods using acids or alkalis. Liquid amino acid fertilizers produced through enzymatic hydrolysis are more advantageous than the chemically produced ones; they are chloride-free, contain a wider range of amino acid components, promote plant growth, and activate beneficial microorganisms in the soil. Furthermore, Liquid amino acid fertilizers are eco-friendly and cost-effective because they reduce the environmental pollution caused by chemical fertilizers and wastewater treatment costs [10]. Peptide, protein-hydrolyzing enzymes, can be divided into exopeptides and endo-peptiddases depending on their cleavage patterns. Exopeptiddases act on the ends of peptide chains and sequentially release amino acids, while endopeptidase at internally by cleaving various peptides. Due to the specificity of these two types of proteases, they can influence the formation of Lysates with various peptide profiles. Accordingly, research on the enzymatic hydrolysis of Livestock blood has primarily focused on its application as a raw material in food additives and pharmaceutical products [11; 12; 13; 14]. While research has been consistently conducted on various types of proteases to produce amino acid fertilizers from different organic waste materials, such as fishery [15] and food waste [16], studies on the production of amino acid fertilizers using Livestock blood as the raw material have not yet been published. Low efficiency and high cost have been pointed out as major disadvantages of protein degradation reactions using enzymes [17; 18].