Amperometric biosensor for lactate analysis in wine and must during fermentation

Claude Bernard University Lyon 1, Villeurbanne, France
Materials Science and Engineering C (Impact Factor: 3.09). 07/2008; 28(5-6):943-948. DOI: 10.1016/j.msec.2007.10.038
Source: OAI


A lactate oxidase-based amperometric biosensor is designed for lactate determination. Two methods of lactate oxidase immobilization on the surface of commercial SensLab platinum printing electrodes are compared. The sensor with lactate oxidase immobilized by physical adsorption in Resydrol polymer is shown to have both narrower dynamic range (0.004–0.5 mМ lactate) and higher sensitivity (320 nA/mM) as compared with that immobilized in poly(3,4-ethylenedioxythiophene) by electrochemical polymerization (0.05–1.6 mM and 60 nA/mM respectively). The operational stability of the biosensors developed is studied; the immobilization method is shown to be of no influence. The lactate content in wine and in wine material during fermentation is analyzed. The data obtained by amperometric lactate biosensor correlated with those of standard chromatography. The biosensor developed can be used in food industry for control and optimization of process of wine fermentation as well as for control of wine quality.

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Available from: Alexei P Soldatkin, Oct 03, 2015
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    • "An important biocompound that is in need of reliable monitoring is l-lactate. Biosensor applications for the determination of l-lactate range from biotechnology (food quality control [13], cell culture monitoring [14]) to monitoring different aspects of human health [15]. Some previous examples for the amperometric biosensing of l-lactate have been described using l-lactate oxidases (FMNdependent ) and l-lactate dehydrogenases (NAD + -dependent) from various natural sources. "
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    ABSTRACT: A comparison between engineered and commercially available l-lactate oxidases from Aerococcus viridans was conducted for biosensing applications. Enzymes were adsorbed onto the surfaces of graphite electrodes modified with multi-walled carbon nanotubes. Thermostable l-lactate oxidases were cloned with a (i) N-, (ii) a C-terminal His-tag and (iii) a wild-type enzyme. Subsequently to the heterologous expression in Escherichia coli and purification, we determined the kinetic parameters of these enzymes in solution. The kinetics of the wild-type, of the N-terminally His-tagged enzyme and of the commercial l-lactate oxidase from A. viridans were studied with a classical Michaelis–Menten as well as with a substrate inhibition model, while the enzyme carrying a C-terminal His-tag showed no activity. The active enzymes were used to fabricate and comparatively investigate multi-walled carbon nanotubes-based biosensors. The enzyme kinetic results were compared with electrochemical studies. By using both spectrophotometric and amperometric techniques, the inhibition phenomenon fits better to the data especially those data related with Lox-His-N. The electrochemical data of the fabricated enzymatic biosensors showed that the N-terminally His-tagged l-lactate oxidase performed best on carboxyl-modified carbon nanotubes. The sensor based on this engineered enzyme showed the highest sensitivity and lowest detection limit in the range of l-lactate concentration 0–1 mM as well as long term stability over one month.
    Electrochimica Acta 03/2013; 93:72-79. DOI:10.1016/j.electacta.2013.01.080 · 4.50 Impact Factor
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    • "Electrochemical polymerization is of remarkable interest due to its technological facilities. It enables to select and maintain dimensions , shape and thickness of the matrix and to provide exact control over precipitation [18] [19]. Poly(3,4-ethylenedioxythiophene) (PEDT) is one of polythiophenes , conductive polymers with novel promising properties. "
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    ABSTRACT: A comparative analysis of the efficiency of using glycerol oxidase preparations, which differ in their characteristics and methods of production, in the development of an amperometric biosensor for glycerol determination, was performed. The enzyme preparation which, being immobilized on the transducer surface, ensured the best working characteristics of the sensor, was selected. Electrochemical polymerization of the selected enzyme preparation in polymer poly(3,4-ethylenedioxythiophene) was chosen as the most effective method of glycerol oxidase immobilization on the surface of amperometric biosensor. pH optimum of the developed amperometric biosensor was determined to be 7.2. Buffer capacity and background electrolyte concentration in buffer solution were shown to have no effect on the work of glycerol biosensor. Glycerol concentration in wine sample solutions was measured using the developed amperometric biosensor based on glycerol oxidase.
    Sensors and Actuators B Chemical 02/2010; 144(2-144):361-367. DOI:10.1016/j.snb.2008.11.051 · 4.10 Impact Factor
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    • "Electrochemical polymerization is of remarkable interest due to its technological facilities. It enables to select and maintain dimensions , shape and thickness of the matrix and to provide exact control over precipitation [18] [19]. "
    Sensors and Actuators B Chemical 01/2010; 144(2):361-367. · 4.10 Impact Factor
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