The Electrochemical Leaf:Rapid, Reversible Cycling of Nicotinamide Cofactors for Enzyme-based Organic Synthesis
Lead Research Organisation:
University of Oxford
Department Name: Mathematical, Physical&Life Sciences Div
Abstract
A new invention, the 'Electrochemical Leaf' integrates a universal component of green leaves with a conducting metal oxide to produce a unique bio-hydrid material to drive enzyme-based organic synthesis. The 'Leaf is a platform technology with numerous applications. The first bond-forming stage of photosynthetic CO2 uptake in plants involves a simple enzyme called ferredoxin-NADP-reductase (FNR) which uses photo-excited electrons to recycle the 'nicotinamide' cofactor NADPH that is consumed (yielding NADP+) in the primary CO2-fixing process (Calvin cycle). In the 'Leaf, FNR is embedded in mesoporous indium tin oxide (ITO) deposited on a support, resulting in an electrode (FNR@ITO/support) that recycles NADPH/NADP+ rapidly and reversibly, using electricity. More than 1000 enzymes use nicotinamides in selective organic reactions, but industry cannot exploit this chemistry. The 'Leaf' is a platform technology that represents a step change.
People |
ORCID iD |
| Fraser Armstrong (Principal Investigator) |
Publications
Armstrong F
(2022)
From Protein Film Electrochemistry to Nanoconfined Enzyme Cascades and the Electrochemical Leaf
in Chemical Reviews
Armstrong F
(2021)
Some fundamental insights into biological redox catalysis from the electrochemical characteristics of enzymes attached directly to electrodes
in Electrochimica Acta
Cheng B
(2022)
Deracemisation and stereoinversion by a nanoconfined bidirectional enzyme cascade: dual control by electrochemistry and selective metal ion activation.
in Chemical communications (Cambridge, England)
Evans RM
(2019)
The value of enzymes in solar fuels research - efficient electrocatalysts through evolution.
in Chemical Society reviews
Herold R
(2023)
Electrochemical Nanoreactor Provides a Comprehensive View of Isocitrate Dehydrogenase Cancer-drug Kinetics
in Angewandte Chemie
Herold R
(2021)
Exploiting Electrode Nanoconfinement to Investigate the Catalytic Properties of Isocitrate Dehydrogenase (IDH1) and a Cancer-Associated Variant
in The Journal of Physical Chemistry Letters
Herold RA
(2023)
Electrochemical Nanoreactor Provides a Comprehensive View of Isocitrate Dehydrogenase Cancer-drug Kinetics.
in Angewandte Chemie (International ed. in English)
Herold RA
(2023)
NADP(H)-dependent biocatalysis without adding NADP(H).
in Proceedings of the National Academy of Sciences of the United States of America
Liu X
(2023)
Natural and synthetic 2-oxoglutarate derivatives are substrates for oncogenic variants of human isocitrate dehydrogenase 1 and 2.
in The Journal of biological chemistry
Megarity C
(2019)
Electrified Nanoconfined Biocatalysis with Rapid Cofactor Recycling
in ChemCatChem
Megarity C
(2019)
Electrocatalytic Volleyball: Rapid Nanoconfined Nicotinamide Cycling for Organic Synthesis in Electrode Pores
in Angewandte Chemie
Megarity CF
(2019)
Electrocatalytic Volleyball: Rapid Nanoconfined Nicotinamide Cycling for Organic Synthesis in Electrode Pores.
in Angewandte Chemie (International ed. in English)
Megarity CF
(2020)
Electron flow between the worlds of Marcus and Warburg.
in The Journal of chemical physics
Morello G
(2019)
Efficient Electrocatalytic CO 2 Fixation by Nanoconfined Enzymes via a C3-to-C4 Reaction That Is Favored over H 2 Production
in ACS Catalysis
Morello G
(2021)
The power of electrified nanoconfinement for energising, controlling and observing long enzyme cascades.
in Nature communications
Reinbold R
(2022)
Resistance to the isocitrate dehydrogenase 1 mutant inhibitor ivosidenib can be overcome by alternative dimer-interface binding inhibitors
in Nature Communications
Wan L
(2018)
A hydrogen fuel cell for rapid, enzyme-catalysed organic synthesis with continuous monitoring.
in Chemical communications (Cambridge, England)
| Description | We have investigated a number of biosynthetic reactions and started a very productive collaboration with Nick Turner (University of Manchester) aimed at revolutionising the study and exploitation of organic synthesis catalysed by redox enzymes. A very important new factor is our ability to monitor, continuously, the progress of the reaction underway, and make interventions to check different aspects and make adjustments. We are examining reductive amination, imine reduction, alcohol/ketone interconversions and carboxylate reduction using the respective 'coupling enzymes'. We have designed reactors in which the process is turned into a fuel cell. In one case, we have used hydrogen to convert 2-ketoglutarate and ammonia into L-glutamate, and used this reaction to study the kinetics of the two-enzyme system (now published 2018). In another case, we use air (passed over a Pt cathode) to convert alcohols into ketones, in many examples with R/S stereoselectivity. In what is a major discovery, we have established that the electrochemical leaf works because of nanoconfinement of two different enzymes between which NADP(H) can be recycled very rapidly. The coupling enzyme (E2) must bind very close to the NADP(H) recycling enzyme FNR, within each of the billions of nanospaces formed by electrophoretic deposition of indium tin oxide (ITO) nanoparticles. The nanoconfinement of FNR, E2 and NADP(H) in the nanozones |(analogous to that represented by a 10 x 10 x 10 nm cube) results in a massive increase in the concentration of each component and minimises the distance that NADP(H) must transfer during recycling. The system is unique in linking cofactor recycling between two enzymes trapped in electrode nanopores, fast and reversible electrochemical driving of NADP(H) via one of the enzymes (FNR), and sophisticated organic transformations by E2, which can be any of hundreds of different oxidoreductase. A patent is now published. We have shown that we now have a method for setting up and controlling extended enzyme cascades, effectively making myriad enzymes directly controllable and observable by electrochemical technology. We are now developing the technology along further fronts: we have starting using titanium foam as the support, which offers a very high surface area. As well as revolutionalising biocatalyst discovery (because real-time observation and control makes this so easy) we are now designing scaled-up reactors that should be proof-of-principle for pharma companies and we have designed a system for deracemizing secondary alcohols. Most recently we are exploiting the technology platform to investigate inhibitors of cancer-associated enzymes. |
| Exploitation Route | The concept of an energising, controlling and monitoring enzyme cascades trapped in electrode nanopore zones is unprecedented. New fundamental studies will include materials, mathematics, enzymology and organic synthesis via biocatalysis. The work is poised to revolutionise enzyme-based organic synthesis because not only are we able to recycle nicotinamide cofactors very rapidly (and for > 40,000 turnovers) but we are also able to monitor the process continuously in real time and make adjustments by simple interventions as required. We have already made good progress with scaling up and have demonstrated a device for the straightforward deracemization of secondary alcohols. Our work will also make it very easy to screen enzymes for desired activities, both in detail and in a fraction of the time currently required by laboratories. We have shown how we can run extended enzyme cascades with control over rate and direction with simultaneous monitoring, and are now demonstrating its application for the study of drugs that inhibit cancer-associated variant enzymes. |
| Sectors | Agriculture, Food and Drink,Chemicals,Energy,Environment,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | The 'Electrochemical Leaf' (e-Leaf) is proving much more powerful than first realised. We are on track to commercialise the technology after having had a patent and more than 10 significant papers published. I have had much more help now from Oxford University Innovation, due to positive intervening by Serena de Nahlik. I am in contact with several companies and a video has been produced. |
| First Year Of Impact | 2017 |
| Sector | Chemicals,Education,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Cultural,Economic |
| Description | BBSRC Follow on Fund |
| Amount | £197,917 (GBP) |
| Funding ID | BB/P023797/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 07/2017 |
| End | 12/2018 |
| Description | Collaboration started with Professor Nick Turner, University of Manchester |
| Organisation | University of Manchester |
| Department | Manchester Institute of Biotechnology MIB |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Turner is supplying enzymes that we are using in our research. We are now writing our first joint paper, after 6 months collaboration. |
| Collaborator Contribution | Supply of various enzymes. |
| Impact | A grant application was submitted in October 2017 to BBSRC (Manchester and Oxford) to develop the science and technology for biocatalyst discovery. |
| Start Year | 2017 |
| Title | ELECTRODES |
| Description | An electrode (1), the electrode (1) comprises a substrate (4, 5) on which is located a porous layer of a conducting or semi-conducting oxide (6) and having located thereon Ferredoxin NADP Reductase (FNR) (3). The electrode (1) can be used to drive organic synthesis via nicotinamide cofactor regeneration. |
| IP Reference | WO2017158389 |
| Protection | Patent application published |
| Year Protection Granted | 2017 |
| Licensed | No |
| Impact | Discovery of a fundamentally new way of driving enzyme cascades for organic synthesis. The discovery is of value for both practical and theoretical reasons. The science is explained in a 2019 'hot' paper in Angewandte Chemie. |
| Description | Invited lecture |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Invited lecture at the Gerischer-Kolb Symposium, Reisenberg Castle, Germany, October 11-13, 2017 |
| Year(s) Of Engagement Activity | 2017 |