An amperometric pyruvate biosensor based on pyruvate oxidase nanoparticles immobilized onto pencil graphite electrode. (June 2020)

• Record Type: Journal Article
• Authors: Malik, Mansi; Chaudhary, Reeti; Pundir, C.S.
• Is Part Of: Process biochemistry. Volume 93(2020)
• Page Start: 12
• Page End: 20
• Publication Date: 2020-06
• Subjects: Pyruvate -- Pyruvate oxidase (POx) -- Pyruvate oxidase nanoparticles (POxNPs) -- Pyruvate biosensor -- Pencil graphite electrode (PGE); Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2020.03.008
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: POxNPs/PGE = Pyruvate oxidase nanoparticles; PGE = Pencil graphite electrode Highlights: Prepared and characterized pyruvate oxidase nanoparticles (POxNPs). Immobilized POxNPs onto pencil graphite electrode to construct improved amperometric pyruvate biosensor. Biosensor displayed limit of detection as 0.58 μM within a linear working range, 0.001–6000 μM. Biosensor measured elevated level of pyruvate in sera of heart patients. Working electrode showed storage stability of 210 days, while being stored dry at 4 °C. Abstract: This present study was aimed to fabricate a sensitive and improved amperometric biosensor by the nanoparticles of pyruvate oxidase, which were prepared and immobilized covalently onto pencil graphite electrode. The biosensor showed ideal working within 5 s under defined conditions of pH 6.0 and incubation temperature of 30 °C at an applied voltage of -0.1 V. Under standard assay conditions, a linear response was obtained between pyruvate concentration ranging from 0.001 to 6000 μM and current (μA). A lower detection limit (0.58 μM) and an excellent correlation coefficient (R 2 = 0.999) with standard spectrophotometric assay was obtained for the present biosensor. Within and between batches of coefficients of variation were calculated and found to be 3.61 % and 3.33 %, respectively. The biosensor was put to continual use for over 210 days. The biosensor was employed for the measurement of pyruvate level in sera of normal healthy individuals and persons suffering from heart disease.