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A Novel Amperometric Glutamate Biosensor Based on Glutamate Oxidase Adsorbed on Silicalite

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ABSTRACT

In this work, we developed a new amperometric biosensor for glutamate detection using a typical method of glutamate oxidase (GlOx) immobilization via adsorption on silicalite particles. The disc platinum electrode (d = 0.4 mm) was used as the amperometric sensor. The procedure of biosensor preparation was optimized. The main parameters of modifying amperometric transducers with a silicalite layer were determined along with the procedure of GlOx adsorption on this layer. The biosensors based on GlOx adsorbed on silicalite demonstrated high sensitivity to glutamate. The linear range of detection was from 2.5 to 450 μM, and the limit of glutamate detection was 1 μM. It was shown that the proposed biosensors were characterized by good response reproducibility during hours of continuous work and operational stability for several days. The developed biosensors could be applied for determination of glutamate in real samples.

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Dependence of responses of biosensor based on GlOx, adsorbed on silicalite, on enzyme concentration at membrane deposition. Measurements in 20 mM HEPES buffer, pH 7.4, at a constant potential of +0.6 V vs Ag/AgCl reference electrode. Glutamate concentration, 1 mM
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Fig6: Dependence of responses of biosensor based on GlOx, adsorbed on silicalite, on enzyme concentration at membrane deposition. Measurements in 20 mM HEPES buffer, pH 7.4, at a constant potential of +0.6 V vs Ag/AgCl reference electrode. Glutamate concentration, 1 mM

Mentions: An effect of the enzyme concentration was more remarkable. Figure 6 shows the experimental results regarding an influence of GlOx concentration during adsorption on the biosensor responses. As it can be seen, an increase of GlOx concentration in HEPES solution from 0.5 to 4% led to almost four-times increase in the biosensor response because of the increased amount of adsorbed GlOx. However, GlOx concentrations higher than 4% did not improve the results, probably because of an excess of the enzyme. Thus, 4% GlOx solution can be considered as optimal for the biosensor creation.Fig. 6


A Novel Amperometric Glutamate Biosensor Based on Glutamate Oxidase Adsorbed on Silicalite
Dependence of responses of biosensor based on GlOx, adsorbed on silicalite, on enzyme concentration at membrane deposition. Measurements in 20 mM HEPES buffer, pH 7.4, at a constant potential of +0.6 V vs Ag/AgCl reference electrode. Glutamate concentration, 1 mM
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5383914&req=5

Fig6: Dependence of responses of biosensor based on GlOx, adsorbed on silicalite, on enzyme concentration at membrane deposition. Measurements in 20 mM HEPES buffer, pH 7.4, at a constant potential of +0.6 V vs Ag/AgCl reference electrode. Glutamate concentration, 1 mM
Mentions: An effect of the enzyme concentration was more remarkable. Figure 6 shows the experimental results regarding an influence of GlOx concentration during adsorption on the biosensor responses. As it can be seen, an increase of GlOx concentration in HEPES solution from 0.5 to 4% led to almost four-times increase in the biosensor response because of the increased amount of adsorbed GlOx. However, GlOx concentrations higher than 4% did not improve the results, probably because of an excess of the enzyme. Thus, 4% GlOx solution can be considered as optimal for the biosensor creation.Fig. 6

View Article: PubMed Central - PubMed

ABSTRACT

In this work, we developed a new amperometric biosensor for glutamate detection using a typical method of glutamate oxidase (GlOx) immobilization via adsorption on silicalite particles. The disc platinum electrode (d = 0.4 mm) was used as the amperometric sensor. The procedure of biosensor preparation was optimized. The main parameters of modifying amperometric transducers with a silicalite layer were determined along with the procedure of GlOx adsorption on this layer. The biosensors based on GlOx adsorbed on silicalite demonstrated high sensitivity to glutamate. The linear range of detection was from 2.5 to 450 μM, and the limit of glutamate detection was 1 μM. It was shown that the proposed biosensors were characterized by good response reproducibility during hours of continuous work and operational stability for several days. The developed biosensors could be applied for determination of glutamate in real samples.

No MeSH data available.


Related in: MedlinePlus