Effects of metal ions on activity and structure of phenoloxidase in Penaeus vannamei

Abstract

Phenoloxidase (PO) is a typical metal enzyme, which requires metal ions as prosthetic groups to enable the full exertion of its activity. To study how metal ions affected the activity and structure of PO enzymes, while providing reference materials for in-depth investigations, we examined the effects of different metal ions (Cu²⁺, Zn²⁺, Mg²⁺, Ca²⁺, and Ba²⁺) on their activities. Furthermore, Cu²⁺ and Mg²⁺ were selected for further investigation through UV spectra, intrinsic fluorescence spectroscopy, AFM, and FTIR. It was revealed that Cu²⁺ had a more obvious effect on PO compared to Mg²⁺. The PO could be activated when the concentrations of Cu²⁺ and Mg²⁺ were lower than 10^-3 and 10^-2 mol/L, respectively, and maximum PO activities (182.14% and 141.02%) were observed at 10^-4 mol/L concentrations of Cu²⁺ and Mg²⁺. When the concentrations of Cu²⁺ and Mg²⁺ were higher than 10^-2 and 10^-1 mol/L, the activities PO were inhibited. The results of the UV-vis and fluorescence spectra revealed that Cu²⁺ shaped the tertiary structure of PO, whereas the effect of Mg²⁺ was slight. The AFM results demonstrated that high concentrations of Cu²⁺ and Mg²⁺ resulted in PO aggregation. FTIR analysis indicated that the total content of PO α-helices and β-sheets decreased with higher concentrations of Cu²⁺ and Mg²⁺.

Keywords: Activity; Metal ions; Penaeus vannamei; Phenoloxidase; Structure.