Self-sufficient heterogeneous biocatalysis systems based on NAD(P) recycling: new perspectives on construction

Liu, Si; Wang, Zhenfu; Wang, Lijie; Zhang, Songping; Sun, Yan

doi:10.1080/01614940.2023.2294215

Catalysis Reviews

2023

DOI: 10.1080/01614940.2023.2294215

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Self-sufficient heterogeneous biocatalysis systems based on NAD(P) recycling: new perspectives on construction

Si Liu,

Zhenfu Wang,

Lijie Wang

et al.

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2024

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Engineering Oxygen-Independent NADH Oxidase Integrated with Electrocatalytic FAD Cofactor Regeneration

Hou,

Yuan,

Dong

et al. 2024

JACS Au

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An electrochemically mediated enzyme process for nicotinamide adenine dinucleotide (NADH) oxidation and biosensing has been developed in which the oxygendependent activities of wild-type NADH oxidase are replaced by electrochemical regeneration of the flavin adenine dinucleotide (FAD) cofactor in the active site. Consequently, the present bioelectrocatalysis does not rely on a continuous oxygen supply through bubbling air or pure oxygen in biosynthetic applications, which reduces enzyme stability. The coupled electrochemical and enzymatic catalysis is achieved through a combination of enzyme immobilization on the electrode and electrochemical oxidation of FADH 2 in the active site mediated by the electron transfer mediator ferrocene carboxylic acid (FcCA). Furthermore, to minimize the effect of dissolved oxygen when the electrocatalytic process is exposed to air, we successfully designed mutations at the Leu40 and Cys42 sites of Leuconostoc mesenteroides (LmNOx) to block the oxygen passage into the active site and to eliminate the native FAD cofactor regeneration half-reaction. The engineered enzymes, whose activities are significantly reduced or inactive in solution, are electrocatalytically active toward conversion of NADH to NAD + , demonstrating successful FAD cofactor regeneration in the active site via electrochemistry. Finally, we developed two highly responsive electrochemical biosensors for NADH detection which has a superior substrate specific to standard detectors using metal electrodes, and comparable detection range and detection limit (1−3 μM).

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Engineering Oxygen-Independent NADH Oxidase Integrated with Electrocatalytic FAD Cofactor Regeneration

Hou,

Yuan,

Dong

et al. 2024

JACS Au

View full text Add to dashboard Cite

show abstract

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Self-sufficient heterogeneous biocatalysis systems based on NAD(P) recycling: new perspectives on construction

Cited by 1 publication

References 145 publications

Engineering Oxygen-Independent NADH Oxidase Integrated with Electrocatalytic FAD Cofactor Regeneration

Engineering Oxygen-Independent NADH Oxidase Integrated with Electrocatalytic FAD Cofactor Regeneration

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