Ultra-sensitive conductometric detection of heavy metals based on inhibition of alkaline phosphatase activity from Arthrospira platensis.

Nadèje Tekaya 1 Olga Saiapina 2 Hatem Ben Ouada 3 Florence Lagarde 4 Hafedh Ben Ouada 5 Nicole Jaffrezic-Renault 4
1 SIMS - Surfaces-(bio)Interfaces - Micro & Nano Systèmes
ISA - Institut des Sciences Analytiques, Laboratoire de Physique & Chimie des Interfaces
2 Laboratory of Biomolecular Electronics
3 INSTM - Institut National des Sciences et Technologies de la Mer
4 SIMS - Surfaces-(bio)Interfaces - Micro & Nano Systèmes
ISA - Institut des Sciences Analytiques
5 Laboratoire de Physique & Chimie des Interfaces
Abstract : This study is based on the conductometric measurement of alkaline phosphatase activity (APA) from the cyanobacterium, Arthrospira platensis, called Spirulina. Cyanobacterium cells were directly immobilized, by physical adsorption, on the ceramic part of gold interdigitated transducers. This activity was inhibited in the presence of heavy metals and a variation of the local conductivity was measured after addition of the substrate. The Michaelis-Menten constant (Km) was evaluated to be 0.75 mM through a calibration curve of the substrate, disodium 4-nitrophenylphosphate p-nitrophenyl phosphate (pNPP). Inhibition of APA was observed with cadmium and mercury with a detection limit of 10(-20) M. The half maximal inhibitory concentration (IC50) was determined at 10(-19) M for Cd(2+) and 10(-17) M for Hg(2+), and the binding affinity of heavy metal (Ki) was equal to the IC50. On the sensor surface, scanning electron microscopy (SEM) images revealed a remarkable evolution of the cyanobacterium's external surface that was attributable to the first defense mechanism against toxic heavy metals in trace. This effect was also confirmed through the important increase of response time τ(90%) recorded for APA response towards the substrate pNPP after cell exposure to metallic cations. Lifetime of the Spirulina-based biosensor was estimated to be more than 25 days.
Keywords : Inhibition APA Conductometric Spirulina Heavy metals BIOSENSOR SENSORS
Document type :
Journal articles
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Submitted on : Wednesday, October 30, 2013 - 1:25:21 PM
Last modification on : Wednesday, April 3, 2019 - 1:30:16 AM

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ISA | INC-CNRS | UNIV-LYON1 | ENS-LYON

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Nadèje Tekaya, Olga Saiapina, Hatem Ben Ouada, Florence Lagarde, Hafedh Ben Ouada, et al.. Ultra-sensitive conductometric detection of heavy metals based on inhibition of alkaline phosphatase activity from Arthrospira platensis.. Bioelectrochemistry, Elsevier, 2013, 90, pp.24-29. ⟨10.1016/j.bioelechem.2012.10.001⟩. ⟨hal-00878619⟩

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