Electroenzymatic Model System for the Determination of Catalytic Activity of Erwinia carotovora L-Asparaginase

Abstract

An electrochemical method for the determination of the catalytic activity of L-asparaginase (ASNase) from Erwinia carotovora was proposed. Our approach is based on the electrooxidation of amino acids from L-asparaginase polypeptide backbones. The electrochemical behavior of ASNase on electrodes obtained by screen-printing modified with single-wall carbon nanotubes (SPE/SWCNTs) as sensing elements demonstrated a broad oxidation peak at 0.5–0.6 V centered at 0.531 ± 0.010 V. We have shown that in the presence of the substrate L-asparagine, the oxidation current of the enzyme was reduced in a concentration-dependent manner. The specificity of electrochemical analysis was confirmed in experiments with glycine, an amino acid with no substrate activity on ASNase and does not reduce the oxidation peak of L-asparaginase. The addition of glycine did not significantly influence the amplitude of the oxidation current. The innovative aspects of the proposed electrochemical sensor are the direct monitoring of ASNase catalytic activity and a reagentless approach, which does not require additional reagents or labels.