Optimization of preparation process and characteristics analysis of porcine hemoglobin modified by enzymatic hydrolysis-phosphorylation

ObjectiveThis study aimed to make high-value use of pig blood and improve the application range of pig hemoglobin (Hb).MethodsHb was modified by enzymolysis and phosphorylation, and its optimum preparation conditions and characteristics were explored. The optimal parameters were determined by response surface test to study the effect of enzymatic hydrolysis-phosphorylation of porcine hemoglobin (HP-Hb).ResultsThe optimal parameters for phosphorylation were 10 g/100 mL phosphate addition, pH 9, 8 g/100 mL pig hemoglobin peptide (pig hemoglobin peptide, H-Hb) addition, and a reaction time of 2.1 hours. Compared with Hb and H-Hb, HP-Hb showed significantly improved emulsification properties and antioxidant activity. Electrophoresis results indicated no significant change in molecular weight. Structural analysis revealed that the phosphate groups were mostly attached to the N atoms. The change of secondary structure of HP-Hb enhanced its structural stability.ConclusionEnzymolysis phosphorylation co-modification significantly improved the emulsification property, antioxidant property and thermal stability of porcine hemoglobin, indicating that co-modification is a feasible method to improve protein.