Enzyme-Nanoparticle Cofactor Regeneration System
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Solution Overview
Problem
Current methods for cofactor regeneration, such as the formate dehydrogenase and glucose dehydrogenase systems, are inefficient and costly, requiring high overpotential and resulting in contamination of products, while electrochemical regeneration at conventional electrodes is energetically inefficient and generates bio-inactive forms of NAD+/NADH.
Innovation Solution
A cofactor regeneration system comprising a first electron transfer component, such as a NADH:acceptor oxido-reductase or NADPH:acceptor oxido-reductase, a second electron transfer component like a hydrogenase moiety or non-biological nanoparticles, and an electronically conducting surface, where these components are immobilized and work together to regenerate cofactors like NADH and NADPH with minimal overpotential and without contaminating products.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If formate dehydrogenase system is used for NADH regeneration, then cofactor regeneration is achieved, but CO2 is produced which strongly affects the pH of the reaction solution and formate contaminates the product
Solution Approach 1:
The patent extracts and eliminates the harmful byproducts (CO2 and formate) from the regeneration system by using a different biochemical pathway. Instead of using formate dehydrogenase which produces CO2 and formate contamination, the invention uses a system that regenerates NADH without generating these harmful substances, thereby removing the source of pH change and contamination.
Solution Approach 2:
The patent introduces an intermediary substance or mechanism to facilitate cofactor regeneration without the harmful effects. By using alternative electron transfer components and intermediaries in the regeneration pathway, the system achieves NADH regeneration while avoiding the direct production of CO2 and formate that would otherwise contaminate the product and alter pH.
2Reliability
If glucose dehydrogenase system is used for NADPH regeneration, then cofactor regeneration is achieved, but glucose and its oxidized form contaminate the product
Solution Approach 1:
The patent removes the source of contamination by replacing the glucose dehydrogenase system with an alternative regeneration pathway that does not use glucose. This extraction of the harmful substrate (glucose) from the system eliminates the contamination issue while maintaining effective NADPH regeneration.
Solution Approach 2:
The patent employs transient or disposable intermediary substances that facilitate regeneration but do not persist in the final product. By using short-lived intermediaries in the electron transfer chain, the system achieves regeneration without leaving contaminating substances in the product, as these intermediaries are rapidly converted or degraded.
3Reliability
If electrochemical regeneration at conventional electrodes is used, then cofactor regeneration is achieved, but large overpotential is required meaning loss of energy
Solution Approach 1:
The patent replaces the electrochemical system (conventional electrodes requiring large overpotential) with a biochemical system using enzymes and electron transfer proteins. This substitution of mechanical/electrical energy with biochemical energy transfer mechanisms eliminates the need for high overpotential while maintaining effective cofactor regeneration through natural enzymatic pathways.
Solution Approach 2:
The patent changes the fundamental parameters of the regeneration system by moving from electrochemical to biochemical mechanisms. By altering the energy transfer parameters from electrical (requiring high overpotential) to biochemical (using enzyme-catalyzed reactions), the system achieves regeneration with minimal energy loss and no large overpotential requirement.
4Loss of energy
If modified electrodes with poly(Neutral Red) are used, then electrochemical regeneration is improved, but modifiers may be toxic or damaging to enzymes
Solution Approach 1:
The patent extracts and removes the harmful poly(Neutral Red) modifier from the electrode system entirely. By taking out this toxic substance, the invention eliminates the risk of enzyme damage while still achieving effective cofactor regeneration through alternative biochemical mechanisms that do not require such modifiers.
Solution Approach 2:
The patent replaces persistent toxic modifiers with transient biochemical intermediaries that perform their function and then degrade or are consumed. These short-lived biochemical intermediaries facilitate electron transfer without remaining as toxic residues that would damage enzymes, thus eliminating the harm while maintaining functionality.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The system provides a highly efficient, rapid, and robust cofactor regeneration with minimal energy loss, allowing for flexible optimization based on specific applications and conditions, and does not require additional soluble reagents, thus avoiding product contamination.
Implementation Method 1
a first electron transfer component selected from one or more polypeptides comprising a NADH:acceptor oxido-reductase or a NADPH:acceptor oxido-reductase
Implementation Method 2
a second electron transfer component selected from a hydrogenase moiety and/or non-biological nanoparticles
Data Source
AI summary
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.