Impedance spectroscopy and conductometric biosensing for probing catalase reaction with cyanide as ligand and inhibitor

International audienceHerein, carboxypeptidase Y (CPY) and thermolysin (TLN) were compared as sensing elements to develop an original biosensor for the direct detection of ochratoxin A (OTA) in olive oil. Electrochemical detection of OTA was performed using biosensors containing either CPY or TLN immobilized through glutaraldehyde vapor cross-linking onto gold interdigitated microelectrodes. Different parameters affecting biosensor sensitivity towards OTA were first optimized. TLN and CPY exhibited an optimal activity under the same conditions: 35 min cross-linking time, working pH of 7 and temperature of 25 degrees C. TLN and CPY biosensors exhibited comparable analytical performances. The conductometric response of both modified electrodes was linear up to 75 mu M. The limits of detection were also very similar (1 mu M for the CPY biosensor and 0.7 mu M for the TLN biosensor). For both enzymes, initial rate vs. OTA concentration plots could be described by hyperbolic curves consistent with the Michaelis-Menten model. K-M(app) and V-max(app), deduced from these curves, were 26 mu M and 3.3 mu S min(-1) respectively for the TLN biosensor. The biosensors signal was reproducible, relative standard deviations calculated from three consecutive measurements of standards in the 1-75 mu M range, being lower than 5%. The signals were also very stable over a 30 days period when biosensors were stored at 4 degrees C in 20 mM phosphate buffer between two measurements. The TLN biosensor was further evaluated for OTA determination in spiked olive oil samples. Recovery values were close to 100% demonstrating the suitability of this method for OTA screening in olive oil. Interestingly, OTA concentrations could be determined without pretreatment of the sample and no matrix effect was observed

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“…38 Evolution of TLN and CPY biosensors response time with OTA concentration was given by Fig. Response time.…”

Section: Analytical Characteristics Of the Biosensorsmentioning

confidence: 99%

“…The response time was previously dened as the time needed to achieve steady-state response DG ss . 38 Evolution of TLN and CPY biosensors response time with OTA concentration was given by Fig. 6.…”

Section: Analytical Characteristics Of the Biosensorsmentioning

confidence: 99%

Comparison of carboxypeptidase Y and thermolysin for ochratoxin A electrochemical biosensing

et al. 2015

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“…Biosensors have potential applications in biotechnology because of their high specificity, sensitivity, and effectiveness. Most immobilized enzymes are used as biosensors (Wilson and Hu, 2000) among them, several dioxygenases and peroxidases are used to detect hydrogen peroxide, phenolic compounds, and metal ions (Bouyahia et al, 2011;Shamsipur et al, 2012).…”

Section: Biosensorsmentioning

confidence: 99%

Bacteria From the Southern Gulf of Mexico: Baseline, Diversity, Hydrocarbon-Degrading Potential and Future Applications

et al. 2021

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The Gulf of Mexico Research Consortium (Consorcio de Investigación del Golfo de México (CIGoM), 2020) was founded in 2015 as a consortium of scientific research and consulting services, specializing in multidisciplinary projects related to the potential environmental impacts of natural and human-induced oil spills in marine ecosystems, to understand and act in the case of possible large-scale oil spills in the Gulf of Mexico. CIGoM comprises more than 300 specialized researchers trained at the most recognized Mexican institutions. Among the main interests of CIGoM are developing the first baseline of the bacterial community inhabiting the southern Gulf of Mexico, investigating the natural degradation of hydrocarbons by bacterial communities and microbial consortia and identifying and characterizing industrially relevant enzymes. In this review, using third-generation sequencing methodologies coupled to function screening methodologies, we report the bacterial profile found in samples of water and sediments in Mexican regions that include the Perdido Fold Belt (northwest of Mexico), Campeche Knolls (in the southeast) and Southwest region of the Gulf of Mexico. We also highlight some examples of novel lipases and dioxygenases with high biotechnological potential and some culturable hydrocarbon-degrading strains used in diverse bioremediation processes.

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“…[31][32][33][34] On the other hand, electrode modification with nanostructure materials is more attractive in biosensing approaches thanks to their excellent properties such as large surface to volume ratio, increasing electron transfer, high surface reaction activity, and strong adsorption. [35][36][37][38][39][40] For example, Attar et al and Wang et al used gold nanoparticles and hydroxyapatite nanowires in the biosensing of cyanide to promote the performance of biosensors, respectively. [6,23] Recently, reduced graphene oxide (rGO) has emerged as a great candidate due to its specific electrical and mechanical properties.…”

Section: Introductionmentioning

confidence: 99%

A sensitive biosensor for cyanide detection using modified glassy carbon electrode with reduced graphene oxide and immobilization of horseradish peroxidase

Ghanavati,

Keshavarz Moraveji,

Iranshahi

2023

Can J Chem Eng

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Cyanide is a highly poisonous and hazardous substance which may release into the environment from natural sources or industrial effluent; therefore, cyanide detection is a fundamental step to prevent environmental pollution and secure health and safety. In this study, we prepared a sensitive amperometric inhibition biosensor for cyanide detection by immobilization of horseradish peroxidase (HRP) enzyme and reduced graphene oxide (rGO) on the surface of glassy carbon electrode (GCE). To do so, we performed the amperometric measurement by modified GCE to test its efficiency in detecting cyanide. The optimum conditions of pH equal to 7.5, −100 mV applied potential, 0.7 μM mediator concentration, and 0.5 mM substrate concentration were found. Then, experiments were performed at different boundary conditions in a range of 0.1 to 10 μM cyanide concentration at optimal conditions and a low detection limit of 0.01 μM was obtained. Also, the possible mechanism of inhibition was analyzed based on the Michalis–Menten equation and non‐competitive inhibition was observed. Due to high sensitivity, low detection limit, and low cost, this biosensor is proposed as a useful method for cyanide determination in real samples.

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Impedance spectroscopy and conductometric biosensing for probing catalase reaction with cyanide as ligand and inhibitor

Cited by 27 publications

References 29 publications

Comparison of carboxypeptidase Y and thermolysin for ochratoxin A electrochemical biosensing

Comparison of carboxypeptidase Y and thermolysin for ochratoxin A electrochemical biosensing

Bacteria From the Southern Gulf of Mexico: Baseline, Diversity, Hydrocarbon-Degrading Potential and Future Applications

A sensitive biosensor for cyanide detection using modified glassy carbon electrode with reduced graphene oxide and immobilization of horseradish peroxidase

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