Investigation of heavy metal effects on immobilized paraoxanase by glutaraldehyde

Abstract

Serum paraoxonase 1 (PON1) was purified from bovine serum using hydrophobic interaction chromotography on Sepharose 4B-coupled l-tyrosine 1-naphthylamine gel, and monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Paraoxonase enzyme was immobilized using different ratios of glutaraldehyde and the maximum activity was observed with 7% glutaraldehyde. The effects of inhibition by Mn⁺², Co⁺² and Cu⁺² heavy metals on the immobilized and free enzyme activities were studied. At the optimum pH and temperature, the K_m and V_max kinetic values for bovine serum paraoxonase and immobilized paraoxonase towards paraoxon substrate were determined as 0.296 × 10⁻³ M & 37.04 EU vs. 0.727–10⁻³ M & 36.36 EU, respectively.

Keywords::

• Paraoxonase
• enzyme immobilization
• glutaraldehyde
• enzyme inhibition

Acknowledgment

This work was carried out in the Balikesir University Research Center of Applied Sciences (BURCAS). The authors would like to thank to Dr. Malcolm Lyon for his invaluable contribution to this paper.

Declaration of interest

The authors report no conflicts of interest.