Purification and structural characterization of a novel antioxidant and antibacterial protein from Arca inflata.

A novel protein (J2-C4) with antioxidant and antibacterial activities was purified from the edible portion of Arca inflata. The purity of J2-C4 was measured to be 97.62% by RP-HPLC analysis, and the molecular weight was 20,537.0 Da by ESI-MS/MS. The isoelectric point of J2-C4 was determined to be 5.18 by IEF-PAGE. Secondary structure analysis of J2-C4 showed that it contained 34.9% α-helix, 15.0% β-sheet, 16.3% β-turn and 34.0% random coil by CD spectroscopy. The complete amino acid sequence of J2-C4 was identified by gel electrophoresis and LC-MS/MS together with transcriptome database analysis. Based on the alignment with NCBI BLAST database, J2-C4 showed 71% homology with a sarcoplasmic calcium-binding protein isolated from Crassostrea virginica. Therefore, J2-C4 was proposed to be a new sarcoplasmic calcium-binding protein-like protein in A. inflata. In vitro antioxidant assays showed that J2-C4 exhibited favorable scavenging activities on ABTS+ (EC50 145.80 μg/mL) and DPPH (EC50 455.62 μg/mL). J2-C4 exhibited antibacterial activities against Pseudomonas aeruginosa (MIC: 375 μg/mL), Escherichia coli (MIC: 187.5 μg/mL) and Methicillin resistant Staphylococcus aureus (MIC: 750 μg/mL). The results showed that J2-C4 might be developed as a potential food additive agent.