Bi-enzyme L-arginine-selective amperometric biosensor based on ammonium-sensing polyaniline-modified electrode

A novel L-arginine-selective amperometric bi-enzyme biosensor based on recombinant human arginase I isolated from the gene-engineered strain of methylotrophic yeast Hansenula polymorpha and commercial urease is described. The biosensing layer was placed onto a polyaniline-Nafion composite platinum electrode and covered with a calcium alginate gel. The developed sensor revealed a good selectivity to L-arginine. The sensitivity of the biosensor was 110 +/- 1.3 nA/(mM mm(2)) with the apparent Michaelis-Menten constant (K-M(app)) derived from an L-arginine (L-Arg) calibration curve of 1.27 +/- 0.29 mM. A linear concentration range was observed from 0.07 to 0.6 mM, a limit of detection being 0.038 mM and a response time - 10 s. The developed biosensor demonstrated good storage stability. A laboratory prototype of the proposed amperometric biosensor was applied to the samples of three commercial pharmaceuticals ("Tivortin", "Cytrarginine", "Aminoplazmal 10% E") for L-Arg testing. The obtained L-Arg-content values correlated well with those declared by producers. (C) 2012 Elsevier B.V. All rights reserved.