Electrochemical characterization of a dual cytochrome-containing lactate dehydrogenase

Boyarski, Anastasya; Shlush, Noam; Paz, Shiraz; Eichler, Jerry; Alfonta, Lital

doi:10.1016/j.bioelechem.2023.108406

Cited by 4 publications

(1 citation statement)

References 35 publications

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“…The first reported Fc b 2 -based ABSs were developed using cells or cell lysates, from the yeasts S. cerevisiae and Hansenula anomala as the parental cells of this enzyme [ 30 , 31 ]. Many Lact-sensitive ABSs based on the recombinant cells have been proposed, especially in the last several years [ 32 , 33 , 34 , 35 , 36 , 37 ].…”

Section: Introductionmentioning

confidence: 99%

Flavocytochrome b2-Mediated Electroactive Nanoparticles for Developing Amperometric L-Lactate Biosensors

et al. 2023

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L-Lactate is an indicator of food quality, so its monitoring is essential. Enzymes of L-Lactate metabolism are promising tools for this aim. We describe here some highly sensitive biosensors for L-Lactate determination which were developed using flavocytochrome b2 (Fcb2) as a bio-recognition element, and electroactive nanoparticles (NPs) for enzyme immobilization. The enzyme was isolated from cells of the thermotolerant yeast Ogataea polymorpha. The possibility of direct electron transfer from the reduced form of Fcb2 to graphite electrodes has been confirmed, and the amplification of the electrochemical communication between the immobilized Fcb2 and the electrode surface was demonstrated to be achieved using redox nanomediators, both bound and freely diffusing. The fabricated biosensors exhibited high sensitivity (up to 1436 A·M−1·m−2), fast responses, and low limits of detection. One of the most effective biosensors, which contained co-immobilized Fcb2 and the hexacyanoferrate of gold, having a sensitivity of 253 A·M−1·m−2 without freely diffusing redox mediators, was used for L-Lactate analysis in samples of yogurts. A high correlation was observed between the values of analyte content determined using the biosensor and referenced enzymatic-chemical photometric methods. The developed biosensors based on Fcb2-mediated electroactive nanoparticles can be promising for applications in laboratories of food control.

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Section: Introductionmentioning

confidence: 99%

Flavocytochrome b2-Mediated Electroactive Nanoparticles for Developing Amperometric L-Lactate Biosensors

et al. 2023

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Visual dual-mode aptasensor for non-small cell lung cancer exosome detection via HRP self-coupling enhanced oxidized iridium nanoparticle aggregation

Xu,

Zhang,

Shen

et al. 2024

ANAL. SCI.

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Peroxidase-like Nanoparticles of Noble Metals Stimulate Increasing Sensitivity of Flavocytochrome b2-Based L-Lactate Biosensors

Gayda,

Demkiv,

Stasyuk

et al. 2024

Biosensors

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We report the development of amperometric biosensors (ABSs) employing flavocytochrome b2 (Fcb2) coupled with nanoparticles (NPs) of noble metals on graphite electrode (GE) surfaces. Each NPs/GE configuration was evaluated for its ability to decompose hydrogen peroxide (H2O2), mimicking peroxidase (PO) activity. The most effective nanoPO (nPO) was selected for developing ABSs targeting L-lactate. Consequently, several Fcb2/nPO-based ABSs with enhanced sensitivity to L-lactate were developed, demonstrating mediated ET between Fcb2 and the GE surface. The positive effect of noble metal NPs on Fcb2-based sensor sensitivity may be explained by the synergy between their dual roles as both PO mimetics and electron transfer mediators. Furthermore, our findings provide preliminary data that may prompt a re-evaluation of the mechanism of L-lactate oxidation in Fcb2-mediated catalysis. Previously, it was believed that L-lactate oxidation via Fcb2 catalysis did not produce H2O2, unlike catalysis via L-lactate oxidase. Our initial research revealed that the inclusion of nPO in Fcb2-based ABSs significantly increased their sensitivity. Employing other PO mimetics in ABSs for L-lactate yielded similar results, reinforcing our hypothesis that trace amounts of H2O2 may be generated as a transient intermediate in this reaction. The presence of nPO enhances the L-lactate oxidation rate through H2O2 utilization, leading to signal amplification and heightened bioelectrode sensitivity. The proposed ABSs have been successfully tested on blood serum and fermented food samples, showing their promise for L-lactate monitoring in medicine and the food industry.

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Electrochemical characterization of a dual cytochrome-containing lactate dehydrogenase

Cited by 4 publications

References 35 publications

Flavocytochrome b2-Mediated Electroactive Nanoparticles for Developing Amperometric L-Lactate Biosensors

Flavocytochrome b2-Mediated Electroactive Nanoparticles for Developing Amperometric L-Lactate Biosensors

Visual dual-mode aptasensor for non-small cell lung cancer exosome detection via HRP self-coupling enhanced oxidized iridium nanoparticle aggregation

Peroxidase-like Nanoparticles of Noble Metals Stimulate Increasing Sensitivity of Flavocytochrome b2-Based L-Lactate Biosensors

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