Assay of dehydrogenases with an O2-consuming biosensor.
Coimmobilization of L-lactate oxidase (LOD) and L-lactate dehydrogenase (LDH) on a film bound to an oxygen electrode makes it possible, by recycling L-lactate and pyruvate, to lower their detection limit to 10 nM. With LOD immobilized alone and LDH in solution, easy LDH measurements were performed. The detection limit was 1 IUl-1. Similarly, this mixed biosensor, using an LDH sensitive to mercurials, was used for p-chloromercuribenzoate (PCMB) detection. Coimmobilization of LOD + LDH with an NAD(+)-dependent dehydrogenase allows the measurements of all dehydrogenases. NAD+ is recycled. The detection limit was 50 nM. Applications can be found in medicine, the food industry, and the environment.
Haouz
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Laboratoire de Technologies des Enzymes et des Biomembranes, Université Paris 7-Denis Diderot, France.
Geloso-Meyer
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Burstein
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C
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D015374:Biosensing Techniques; D002470:Cell Survival; D002729:Chloromercuribenzoates / Q000032:analysis; D004566:Electrodes; D004579:Electron Transport; D004800:Enzymes, Immobilized; D004867:Equipment Design; D004926:Escherichia coli / Q000201:enzymology; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism; D007773:Lactates / Q000032:analysis / Q000378:metabolism; D019344:Lactic Acid; D006899:Mixed Function Oxygenases / Q000032:analysis / Q000378:metabolism; D009243:NAD / Q000032:analysis / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000032:analysis*; D010100:Oxygen; D011773:Pyruvates / Q000032:analysis / Q000378:metabolism; D019289:Pyruvic Acid; D012680:Sensitivity and Specificity; D020245:p-Chloromercuribenzoic Acid
D001426:Bacterial Proteins; D002729:Chloromercuribenzoates; D004800:Enzymes, Immobilized; D007773:Lactates; D011773:Pyruvates; D009243:NAD; D019344:Lactic Acid; D020245:p-Chloromercuribenzoic Acid; D019289:Pyruvic Acid; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; D007770:L-Lactate Dehydrogenase; C021497:lactate 2-monooxygenase; D010100:Oxygen