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

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• 作者：Nataliya Stasyuk^a ; Oleh Smutok^b ; Galina Gayda^b ; Bohdan Vus^c ; Yevgen Koval'chuk^a ; Mykhailo Gonchar^b ; ^d ; ^{gonchar@cellbiol.lviv.ua}
• 关键词：Amperometric biosensor ; l-arginine ; Polyaniline ; Arginase I ; Urease
• 刊名：Biosensors and Bioelectronics
• 出版年：2012
• 出版时间：August-September, 2012
• 年：2012
• 卷：37
• 期：1
• 页码：46-52
• 全文大小：395 K

文摘

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¡À1.3 nA/(mM mm²) with the apparent Michaelis-Menten constant (K_M^app) derived from an l-arginine (l-Arg) calibration curve of 1.27¡À0.29 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.