NADH-dependent formate dehydrogenase mutants for efficient carbon dioxide fixation

Xue, Yaju; Ji, Xiuling; Li, Zhuang; Ma, Fuqiang; Jiang, Jingjie; Huang, Yuhong

doi:10.1016/j.biortech.2023.130027

“…The first type includes enzymes without a metal active center (NADH-dependent FDH). 18 This type of FDH usually exists in plants, yeast, and some aerobic bacteria and uses NADH as a cofactor for CO 2 reduction. Figure 1 summarizes crystal structures, the type of residues in the catalytic center of FDH from Candida boidinii (CbFDH), and the mechanism of CO 2 RR catalyzed by this enzyme at pH ∼7.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Biocatalysts, such as enzymes and microbes, are of particular interest in the CO 2 RR due to their high selectivity and activity toward desired products under mild conditions. , Formate dehydrogenases (FDH) are redox enzymes, which specifically catalyze the reversible conversion between CO 2 and formate (or oxidation of formate to CO 2 ). , There are two types of FDH. The first type includes enzymes without a metal active center (NADH-dependent FDH) . This type of FDH usually exists in plants, yeast, and some aerobic bacteria and uses NADH as a cofactor for CO 2 reduction.…”

Section: Introductionmentioning

confidence: 99%

Electrocatalysis of CO₂ Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

Su,

Elmahdy,

Biernat

et al. 2024

Langmuir

1

0

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Nicotinamide adenine dinucleotide-dependent formate dehydrogenase from Candida boidinii was immobilized in a 1,2-dimyristoyl-sn-glycero-3-phosphocholine/cholesterol floating lipid bilayer on the gold surface as a biocatalyst for electrochemical CO2 reduction. We report that, in contrast to common belief, the enzyme can catalyze the electrochemical reduction of CO2 to formate without the cofactor protonated nicotinamide adenine dinucleotide. The electrochemical data indicate that the enzyme-catalyzed reduction of CO2 is diffusion-controlled and is a reversible reaction. The orientation and conformation of the enzyme were investigated by surface-enhanced infrared reflection absorption spectroscopy. The α-helix of the enzyme adopts an orientation nearly parallel to the surface, bringing its active center close to the gold surface. This orientation allows direct electron transfer between CO2 and the gold electrode. The results in this paper provide a new method for the development of enzymatic electrocatalysts for CO2 reduction.

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A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

Li,

Fu,

Chen

et al. 2024

Angewandte Chemie

0

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Industrial fermentation applications typically require enzymes that exhibit high stability and activity at high temperatures. However, efforts to simultaneously improve these properties are usually limited by a trade‐off between stability and activity. This report describes a computational strategy to enhance both activity and thermal stability of the mesophilic organophosphate‐degrading enzyme, methyl parathion hydrolase (MPH). To predict hotspot mutation sites, we assembled a library of features associated with the target properties for each residue and then prioritized candidate sites by hierarchical clustering. Subsequent in silico screening with multiple algorithms to simulate selective pressures yielded a subset of 23 candidate mutations. Iterative parallel screening of mutations that improved thermal stability and activity yielded, MPHase‐m5b, which exhibited 13.3 °C higher Tm and 4.2 times higher catalytic activity than wild‐type (WT) MPH over a wide temperature range. Systematic analysis of crystal structures, molecular dynamics (MD) simulations, and Quantum Mechanics/Molecular Mechanics (QM/MM) calculations revealed a wider entrance to the active site that increased substrate access with an extensive network of interactions outside the active site that reinforced αβ/βα sandwich architecture to improve thermal stability. This study thus provides an advanced, rational design framework to improve efficiency in engineering highly active, thermostable biocatalysts for industrial applications.

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A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

Li,

Fu,

Chen

et al. 2024

Angew Chem Int Ed

1

0

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Industrial fermentation applications typically require enzymes that exhibit high stability and activity at high temperatures. However, efforts to simultaneously improve these properties are usually limited by a trade‐off between stability and activity. This report describes a computational strategy to enhance both activity and thermal stability of the mesophilic organophosphate‐degrading enzyme, methyl parathion hydrolase (MPH). To predict hotspot mutation sites, we assembled a library of features associated with the target properties for each residue and then prioritized candidate sites by hierarchical clustering. Subsequent in silico screening with multiple algorithms to simulate selective pressures yielded a subset of 23 candidate mutations. Iterative parallel screening of mutations that improved thermal stability and activity yielded, MPHase‐m5b, which exhibited 13.3 °C higher Tm and 4.2 times higher catalytic activity than wild‐type (WT) MPH over a wide temperature range. Systematic analysis of crystal structures, molecular dynamics (MD) simulations, and Quantum Mechanics/Molecular Mechanics (QM/MM) calculations revealed a wider entrance to the active site that increased substrate access with an extensive network of interactions outside the active site that reinforced αβ/βα sandwich architecture to improve thermal stability. This study thus provides an advanced, rational design framework to improve efficiency in engineering highly active, thermostable biocatalysts for industrial applications.

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NADH-dependent formate dehydrogenase mutants for efficient carbon dioxide fixation

Cited by 5 publications

References 27 publications

Electrocatalysis of CO₂ Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

Electrocatalysis of CO₂ Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

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NADH-dependent formate dehydrogenase mutants for efficient carbon dioxide fixation

Cited by 5 publications

References 27 publications

Electrocatalysis of CO2 Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

Electrocatalysis of CO2 Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

A Dual‐Focus Workflow for Simultaneously Engineering High Activity and Thermal Stability in Methyl Parathion Hydrolase

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Product

Resources

About

Electrocatalysis of CO₂ Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center

Electrocatalysis of CO₂ Reduction by Immobilized Formate Dehydrogenase without a Metal Redox Center