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Updated: Aug 5, 2026

A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
Published on: August 17, 2019
A simple strategy for enhancing NADPH-dependent biocatalysis via phosphate analogue-mediated maintenance of the NADPH
Rong Liu1,2, Jialing Liu1, Renmei Zhang1
1Department of Biochemistry, School of Preclinical Medicine, Zunyi Medical University Zunyi 563000 Guizhou China yangjw@zmu.edu.cn.
Abstract:
NADPH-dependent biocatalysis is widely used for the synthesis of valuable chemicals, and maintaining an effective NADPH pool during catalysis is critical for its performance. Although cofactor regeneration has been extensively studied, the maintenance of the NADPH pool in complex biocatalytic environments, such as crude-enzyme and whole-cell systems, has received comparatively limited attention. Herein, we found that phosphate analogue additives improved NADPH maintenance in crude-enzyme and whole-cell biocatalytic systems, thereby improving catalytic efficiency. Improved NADPH maintenance was observed in both GDH- and FDH-based regeneration systems. In multiple catalytic systems, including reductase-catalysed reactions, monooxygenase-catalysed reactions, a redox deracemisation cascade, and a multienzyme whole-cell system, catalytic performance was improved by up to approximately 6-fold. These results highlight that maintaining the effective NADPH pool can serve as a simple and cost-effective strategy to improve NADPH-dependent biocatalysis in complex catalytic environments.
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