OPTIMIZATION AND CHARACTERIZATION OF BIOACTIVE PEPTIDES FROM MILK AND FISH PROTEINS USING RESPONSE SURFACE METHODOLOGY
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Abstract
Bioactive peptide have a growing interest in nutraceutical and functional food applications, as they possess antioxidant and therapeutic properties. The current research paper entailed the extraction of the water-soluble peptides (WSPs) in buffalo milk casein, whey, and tuna ( Thunnus albacares ) fish bones, optimization of the process, and characterization of the structure. Alcalase, trypsin, and chymotrypsin enzymatic hydrolysis was performed under controlled physicochemical conditions, and optimization of the hydrolysis parameters was done based on the Response Surface Methodology (Central Composite Design), where Degree of Hydrolysis (DH) was taken as the chief ratio. The optimum conditions varied with the protein sources, which is substrate-specific hydrolytic behavior. The ensuing lyophilized WSPs were fractionated and measured by RP-HPLC, which had a characteristic chromatographic pattern in peptide sources. Mixed WSPs (1:1:1) were associated with the highest peptide concentration with the major peaks eluting at 18-22 min whereas whey WSPs exhibited the least intensity of the peaks. Amino acid profiling showed that there were 17 amino acids and mixed WSPs had high concentrations of essential amino acids, especially phenylalanine and lysine as well as the hydrophobic amino acids including proline and alanine, which indicated better biofunctional potential. The evaluation of antioxidant properties showed that there was significant dose-related enhancement of radical scavenging ability with mixed WSPs possessing much higher DPPH scavenging ability (39.61+ -0.40% at 800 mg/mL) and FRAP values (1.89+-0.40 mM Fe2+/g) than the respective individual sources of WSP. These results suggest a possibility of synergistic effects of combinatorial peptide fractions to increase antioxidant potential. In general, the research defines optimal conditions of extraction of milk- and fish-derived peptides and the high antioxidant capacity of the mixed WSPs, which justifies their use in functional food, nutraceutical, and biologically active ingredients.
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Sana Mahmood
National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan 38000.
Aysha Sameen
Food Sceince and Technology, GC Women University Faisalabad, Pakistan 38000
National Institute of Food Science and Technology, University of Agriculture, Faisalabad.
Muhammad Issa Khan
National Institute of Food Science and Technology, University of Agriculture, Faisalabad
Muhammad Naeem Faisal
Institute of Physiology and Pharmacology, University of Agriculture, Faisalabad, Pakistan 38000.