Abstract
Background: Bioactive peptides from κ-casein have immense therapeutic potential as prophylactic formulations. Among these, casoplatelin is a κ-casein derived bioactive peptide with anti-thrombotic activities. Aim: Herein, we report the production of casoplatelin in an E. coli expression system (using a pBAD vector) and show in silico modeling of its interactions. Methods: A synthetic DNA construct encoding casoplatelin was designed with pepsin cleavage sites before and after the synthetic construct to allow the release of the peptide from the pro-peptide. Results: A novel recombinant approach was demonstrated for the production of casoplatelin, and anti-platelet aggregation activities of the product were confirmed. Also, casoplatelin structures were characterized in silico and then implemented to determine potential structural interactions with fibrinogen. Conclusion: The present study showcases the recombinant approach for biopeptide production and its interaction with fibrinogen through in silico approach.
Highlights
Bioactive Peptides (BAP) constitute milk protein fragments with various reported therapeutic activities, including gastrointestinal, immunological, and nutritional effects [1]
The present study showcases the recombinant approach for biopeptide production and its interaction with fibrinogen through in silico approach
The codon frequency table of E. coli was consulted for each amino acid of casoplatelin, and the DNA sequence was designed with two pepsin cleavage sites flanking the synthetic construct
Summary
Bioactive Peptides (BAP) constitute milk protein fragments with various reported therapeutic activities, including gastrointestinal, immunological, and nutritional effects [1]. BAPs are derived from animal and plant caseins, Overlapping peptide sequences of milk-derived peptides exerts various biological responses, such as ACE inhibition by β-casomorphins and β-lactorphin, immunostimulating effects. Casein glycomacropeptides (GMP) are found in sweet whey, and can be derived from the κ-casein using chymosin or trypsin. The carbohydrate portions N-acetylneuraminic acid (NANA) and N-acetylgalactosamine, and the peptide portions of κ-casein, are responsible for the biological effects of GMP. Mechanisms of carbohydrate-based biological responses are unknown, specific GMP peptides have distinct mechanisms for their respective biological functions [6]. Bioactive peptides from κ-casein have immense therapeutic potential as prophylactic formulations. Casoplatelin is a κ-casein derived bioactive peptide with anti-thrombotic activities
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