Biosensor for rapid determination of 3-hydroxybutyrate using bi-enzyme system.
Biosens Bioelectron
; 21(7): 1101-6, 2006 Jan 15.
Article
en En
| MEDLINE
| ID: mdl-15886000
ABSTRACT
A bi-enzyme-based Clark electrode was developed for the determination of 3-hydroxybutyrate. This sensor is based on the specific dehydrogenation by 3-hydroxybutyrate dehydrogenase (HBDH, E.C. 1.1.1.30) in combination with salicylate hydroxylase (SHL E.C. 1.14.13.1). The enzymes were entrapped by a poly(carbamoyl) sulfonate (PCS) hydrogel on a Teflon membrane. The principle of the determination scheme is as follows the specific detecting enzyme, HBDH, catalyses the specific dehydrogenation of 3-hydroxybutyrate consuming NAD(+). The products, NADH, initiate the irreversible decarboxylation and the hydroxylation of salicylate by SHL in the presence of oxygen. SHL forces the equilibrium of dehydrogenation of 3-hydroxybutyrate by HBDH to the product side by consuming NADH. Dissolved oxygen acts as an essential material for SHL during its enzymatic reactions. This results in a detectable signal due to the SHL-enzymatic consumptions of dissolved oxygen in the measurement of 3-hydroxybutyrate. Interferences from different amino acids and electroactive substances were found to be minimal due to the specificity of HBDH and the application of a Teflon membrane. The sensor has a fast response (2s) and short recovery time (2 min) with a linear range between 8 and 800 microM 3-hydroxybutyrate and a detection limit of 3.9 microM. A good agreement (R(2)=0.9925) with theoretical calculation was obtained in spiked serum sample measurements.
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Bases de datos:
MEDLINE
Asunto principal:
Transductores
/
Técnicas Biosensibles
/
Ácido 3-Hidroxibutírico
/
Electroquímica
/
Hidroxibutirato Deshidrogenasa
/
Oxigenasas de Función Mixta
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
Biosens Bioelectron
Asunto de la revista:
BIOTECNOLOGIA
Año:
2006
Tipo del documento:
Article
País de afiliación:
China