Development of a highly sensitive chemiluminescence flow-injection analysis sensor for phosphate-ion detection using maltose phosphorylase

• Published in: Journal of biotechnology 1999-10, Vol.75 (2), p.127-133
• Main Authors: Nakamura, Hideaki, Hasegawa, Mami, Nomura, Yoko, Arikawa, Yoshiko, Matsukawa, Ritsuko, Ikebukuro, Kazunori, Karube, Isao
• Format: Article
• Language: English
• Subjects: Arthromyces ramosus peroxidase; Biological and medical sciences; Biosensors; Biotechnology; Chemiluminescence; Enzyme immobilization; Enzyme kinetics; Flow-injection analysis; Fundamental and applied biological sciences. Psychology; Methods. Procedures. Technologies; N-Hydroxysuccinimide beads; Organic acids; Phosphate sensor; Phosphates; Various methods and equipments
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/S0168-1656(99)00150-9

A chemiluminescence flow-injection analysis biosensor has been constructed for phosphate-ion detection. This system is coenzymeless and employs a maltose phosphorylase, mutarotase, and glucose-oxidase (MP–MUT–GOD) reaction system combined with an Arthromyces ramosus peroxidase–luminol reaction system. The system consists of a column packed with MP–MUT–GOD immobilized on N-hydroxysuccinimide beads, a mixing joint for the chemiluminescence reaction, and a photomultiplier. The response provided by this system was linear, with a wide range between 10 nM and 30 μM phosphate ion, and a measuring time of 3 min per sample. Under the optimal condition, the sensor was able to detect 1.0 μM phosphate ion for at least 2 weeks. For a practical application, the determination of phosphate ion in river water was examined using ethylenediaminetetraacetic acid, and the results were estimated by comparing with the molybdenum-blue method.