Abstract
An improved amperometric pyruvate biosensor was fabricated by immobilizing covalently commercial pyruvate oxidase (POx) from Aerococcus sps. onto nanocomposite of c-MWCNT (carboxylated multi- walled carbon nanotubes), copper nanoparticles (CuNPs) and polyaniline (PANI) electrodeposited onto gold (Au) electrode. The copper nanoparticles were prepared by chemical reduction method and characterized by transmission electron microscopy (TEM), UV- visible spectroscopy and X- ray diffraction (XRD). The working electrode (POx/c-MWCNT/CuNPs/PANI/AuE was studied via scanning electron microscopy (SEM), Fourier transform infrared spectrometry (FTIR) and electrochemical impedance spectroscopy (EIS) at different stages of its construction. The biosensor showed optimum activity at a pH of 5.0 and 35°C and a linearity for pyruvate in the concentration range, 0.1 µM to 2000 µM. The analytical recovery of added pyruvate was 99.6% and 99.2%. The within and between batch coefficients of variation (CV) were 0.052% and as 0.022% respectively. There was a commendable correlation between sera pyruvate values as measured by standard spectrophotometric method and the present method. The biosensor was applied to measure sera pyruvate level and compared with that biosensor based on pyruvate oxidase nanoparticles covalently bound to Au electrode.
 Malik et al. (2019) Int. J. Appl. Sci. Biotechnol. Vol 7(2): xxx-xxx
 Graphic Abstract
 Schematic representation of fabrication of POx/c-MWCNTs/CuNPs/PANI/AuE and electrochemical reactions involved in its response measurement (POx- Pyruvate oxidase; c-MWCNTs- Carboxylated multi-walled carbon nanotubes; CuNPs- Copper nanoparticles; PANI- Polyaniline; AuE- Gold electrode).
Highlights
Pyruvate is one of the important intermediary molecule holding an important position in several metabolic processes of human body like glycolysis, tricarboxylic acid cycle, lactogenesis and other metabolic pathways (Situmorang et al, 2002)
Characterization of Copper Nanoparticles The Transmission electron microscope (TEM) images of the prepared copper nanoparticles revealed their tiny, round and spherical shape having a mean diameter of 19.70 nm and particle size lying between the range, 15.21 nm to 22.81 nm (Fig. 1a)
A uniform, smooth and featureless pictorial view was obtained by analyzing the bare electrode via Scanning electron microscope (SEM), which upon subsequent deposition with several nanomaterials resulted into the change in the morphology
Summary
Pyruvate is one of the important intermediary molecule holding an important position in several metabolic processes of human body like glycolysis, tricarboxylic acid cycle, lactogenesis and other metabolic pathways (Situmorang et al, 2002). Pyruvate is involved in different energy production pathways, its presence is widely observed in living organisms. The biosynthesis of pyruvate occurs in the cells of the human body (Akylimaz and Yorganci, 2008). Over the last 20 years, several research groups have focused research on developing a novel platform for the detection of pyruvate, as it possesses
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