Structure-based modelling of hemocyanin allergenicity in squid and its response to high hydrostatic pressure

The secondary, tertiary, and quaternary structures of squid hemocyanin (Hc) were characterised, and the relationship between Hc structure and allergenicity responses to high hydrostatic pressure (HHP) was modelled. The Hc allergenicity varied with its protein structure. Electrophoresis analysis revealed that HHP treatment significantly decreased the band intensity of Hc when increasing pressure from 200 and 400 MPa to 600 MPa. The protein structure analysis of squid Hc showed that while HHP treatment decreased the α-helix content, free sulfhydryl content, and R(g), it increased the random coil content, surface hydrophobicity index (Ho), Guinier aggregation number (〈N(agg)〉(G)) and average aggregation number (〈N(agg)〉(Q)). The α-helix and random coil contents of the 600 MPa treated samples were 23.67% and 37.54%, respectively, compared to 32.37% and 32.02% in the control, respectively. HHP treatment decreased the IgE and IgG-binding capacities, indicating a significant decrease in the allergenicity (P< 0.05) of squid Hc. This study provided meaningful information of applying HHP to reduce allergenicity, and explained the responses of Hc protein structure to HHP for lowering the allergenicity of squid.