Physicochemical Properties and Stability of Antioxidant Peptides from Swim Bladder of Grass Carp (Ctenopharyngodon idella)

Grass carp swim bladder collagen peptides (GCPs) were purified by ultrafiltration and Sephadex G-15 chromatography to obtain GCP-II, which exhibited superior antioxidant activity. GCP-II exhibited 78.22% ABTS⁺ scavenging activity and 72.91% Fe²⁺ chelating activity, along with around 90% thermal stability between 4 °C and 100 °C. Environmental factors such as 4% NaCl reduced superoxide scavenging to 59.16% of the original and 0.2% citric acid reduced it to 71.57% of the original, while glucose showed minimal impact on the antioxidant activity of GCP-II. From 464 GCP-II sequences analyzed via LC-MS/MS, 7 bioactive peptides were selected based on antioxidant activity and functional sequence motifs, and were named peptides 1 to 7, respectively. These peptides were synthesized through solid-phase methods, validated for purity exceeding 95% using HPLC and mass spectrometry, and tested for antioxidant performance. Peptides 1, 3, 6, and 7 demonstrated notable antioxidant efficacy: peptide 6 showed 63.31% ABTS⁺ scavenging activity at 1 mg/mL, while peptides 3 and 6 exhibited synergistic effects in DPPH and hydroxyl radical scavenging experiments, surpassing theoretical values by 0.88% and 2.16%, respectively. This study underscores the potential of synthetic GCPs and GCP-II-derived peptides as functional antioxidants, particularly peptide 3 and peptide 6.