Cofactor Regeneration System

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This patent describes a modular cofactor regeneration system for efficiently generating and regenerating biological cofactors such as NADH, NAD+, NADPH, and NADP+. The system utilizes two electron transfer components—one being a specific oxidoreductase (such as a diaphorase derived from different sources or variants thereof) and the other being a hydrogenase or non-biological nanoparticles—both immobilized on an electronically conducting surface (such as carbon materials or metals). This setup enables rapid, energy-efficient electron transfer for cofactor recycling, overcoming major limitations of traditional regeneration approaches.

Use CasesContent extracted from patent full text and abstract with AI.

• Industrial biocatalysis processes where continuous regeneration of cofactors is needed, for example in the pharmaceutical, agrochemical, and fine chemical industries.
• Production of high-value chemicals or pharmaceuticals via enzyme-catalyzed synthesis (e.g., alcohol dehydrogenase or P450 mono-oxygenase reactions).
• Bioreactors for large-scale enzymatic processes requiring cofactor recycling.
• Research applications in synthetic biology where efficient in vitro or cell-free cofactor cycling is required.
• Environmental bioremediation or biosensing systems using oxidoreductase enzymes.
• Development of enzyme kits for laboratory and industrial applications where reliable cofactor supply is essential.

BenefitsContent extracted from patent full text and abstract with AI.

• Greatly increases the efficiency and speed of cofactor regeneration compared to existing biochemical or electrochemical methods.
• Reduces or eliminates contamination from by-products typically produced in standard cofactor recycling systems (e.g., formate or glucose residues).
• Minimizes energy loss by operating near the thermodynamic potential (very low overpotential), making it more energetically efficient.
• Highly modular and customizable—the electron transfer components and surface material can be tuned for specific enzymes, applications, or desired conditions (e.g., oxygen tolerance).
• Improved robustness and operational stability of the system, enabled by covalent or non-covalent immobilization on durable surfaces.
• Facilitates easy separation and reuse of the regeneration particles or surfaces, supporting process scale-up and cost effectiveness.
• Eliminates the need for additional reactants, as systems utilizing H2 avoid introducing difficult-to-remove soluble waste.
• Supports efficient operation across a wider range of conditions than traditional systems, expanding applicability.

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Patent Abstract

The present invention relates to cofactor regeneration systems, components and uses thereof and methods for generating and regenerating cofactors. The cofactor regeneration system comprises a first electron transfer component selected from a polypeptide comprising a NADH:acceptor oxido-reductase or NADPH:acceptor oxido-reductase, a second electron transfer component selected from a hydrogenase moiety and/or non-biological nanoparticles and an electronically conducting surface. The first and second electron transfer components are immobilised on the electrically conducting surface, and the first and second electron transfer components do not occur together in nature as an enzyme complex.