New routes to driving enzyme-catalysed chemical synthesis using hydrogen gas

Lead Research Organisation: University of Oxford
Department Name: Oxford Chemistry

Abstract

Bacterial cells act as miniature chemical factories and have evolved specialised routes to making many of the sorts of molecules that we use as pharmaceuticals, pleasant fragrances, food additives or chemicals for use in agriculture. The key parts of the cells for carrying out this work are the enzymes. It is possible to break cells open and isolate an enzyme for making a specific molecule. Bacteria can also be engineered to make artificial chemicals, expanding the range of molecules they can produce. Procedures are now well-established for growing bacteria on a large scale and isolating large quantities of enzymes, and at the same time, chemical companies are starting to realise the benefits of using enzymes instead of traditional chemical routes. In the production of complicated molecules such as drugs, fragrances and food flavourings, enzymes generate much less waste, make purer chemical products, and allow chemistry to be carried out in water rather than toxic, polluting solvents. The purity of the end product is particularly important in the food and pharmaceutical industries where contaminants may have serious, harmful effects.

Although there has been increasing interest in using enzyme catalysis in chemical production, many challenges remain to be overcome before this approach can be widely adopted. Once isolated from their cells, enzymes are often quite unstable. Their stability can be improved by attaching them to surfaces, but this often requires complicated attachment processes and can be expensive. Secondly, many enzymes only work in the presence of special helper-molecules called cofactors which are used up by the enzymes in the process of making chemicals. The cofactors are also expensive, and for enzyme processes to be economically viable, it is essential to have some way of recycling the cofactors. Unfortunately, the currently-available methods for recycling the cofactors make even more waste which contaminates the desired chemical products. We have developed a technology that addresses both of these challenges, offering a much-needed step change for enzyme catalysis. At the moment, our technology has only been demonstrated on a small scale in our laboratories, but we now need to convince the chemical, pharmaceutical and food industries that this offers real benefits for the future of chemical production.

Our technology works as follows: once we have isolated enzymes from the bacterial cells, we immediately attach them onto cheap carbon beads. This is a very simple one-step process. We attach several different types of enzyme to each bead so that the enzymes can work together to carry out each step in making chemicals. We supply the beads with low, safe levels of hydrogen gas, and this provides the energy for recycling the cofactors and to drive the enzyme machinery necessary to make the required chemicals. To make a desired chemical, we start by putting a cheap chemical building-block in water, we bubble in a little hydrogen gas, add our enzyme-modified beads, and after a few hours, the desired chemical product is ready to collect! The enzyme-modified beads can be easily scooped out of the reaction mixture, leaving nothing else except the desired product and a tiny trace of the harmless cofactor. As an added bonus, the beads can be collected and re-used a number of times, minimising the cost of using enzymes.

To take our concept from a lab-scale idea to a technology ready for industry to adopt, we need to show that we can produce the enzymes on a large scale. We need to show how quickly the beads can produce chemicals, and how pure the products are. This project will answer these sort of questions, so that at the end of the 5 years, we can convince potential customers (chemical, pharmaceutical and food additive companies) that our technology will allow them to make chemicals more cheaply and in a more environmentally-friendly way.

Planned Impact

As described in proposal submitted to IUK
 
Description The grant supports translation towards commercialisation of a biocatalysis technology for fine chemical synthesis. We have made significant progress towards de-risking this technology, for example showing that the enzyme-modified HydRegen catalyst particles can be coupled with a range of different NADH dependent enzymes, operated over a wide pH range, and stored and transported readily.
Exploitation Route We are exploring commercialisation options, including licensing or a spin out.
Sectors Agriculture, Food and Drink,Chemicals,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

URL http://vincent.chem.ox.ac.uk/hydregen.htm
 
Description BBSRC INDUSTRIAL CASE (iCASE) STUDENTSHIP, Interdisciplinary Bioscience DTP, University of Oxford
Amount £80,000 (GBP)
Funding ID Project 1946990, related to BB/M011224/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2017 
End 08/2021
 
Description BBSRC INDUSTRIAL CASE (iCASE) STUDENTSHIP, University of Oxford
Amount £80,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2018 
End 09/2022
 
Description Biocatalysis for Sustainable Chemistry - Understanding Oxidation/Reduction of Small Molecules by Redox Metalloenzymes via a Suite of Steady State and Transient Infrared Electrochemical Methods
Amount € 2,980,000 (EUR)
Funding ID ERC-2018-COG 819580 
Organisation European Research Council (ERC) 
Sector Public
Country European Union (EU)
Start 03/2019 
End 02/2023
 
Description Public Engagement with Research Undergraduate Bursary Award for Summer Project
Amount £2,000 (GBP)
Organisation University of Oxford 
Sector Academic/University
Country United Kingdom
Start 06/2018 
End 08/2018
 
Title Protein Film Infrared Electrochemistry 
Description The method couples direct electrochemical control over electrode-immobilised redox enzymes ('protein film electrochemistry') with infrared spectroscopy to study aspects of the mechanistic of redox enzymes under catalytic turnover conditions. 
Type Of Material Improvements to research infrastructure 
Year Produced 2015 
Provided To Others? Yes  
Impact We have been able to employ this in a series of related publications to understand aspects of the mechanism of NiFe hydrogenases. 
 
Description Cofactor dependent biocatalysis for chemical synthesis 
Organisation Johnson Matthey
Country United Kingdom 
Sector Private 
PI Contribution We have tested a number of dehydrogenase enzymes from Johnson Matthey in conjunction with our H2-driven approach to NADH recycling; we have explored an inherent NAD+ recycling activity present in a number of samples of dehydrogenases.
Collaborator Contribution Johnson Matthey Catalysis and Chiral Technologies have supplied us with a large number of enzyme samples in kind. These include commercially available enzymes as well as newer enzymes from the JMCCT collection which are not yet commercially available.
Impact Enzyme provided in kind by JMCCT contributed to the following publications: (1) Zor, C., Reeve, H.A., Quinson, J., Thompson, L.A., Lonsdale, T.H., Dillon, F., Grobert, N., Vincent K.A., 'H2-driven Biocatalytic Hydrogenation in Continuous Flow using Enzyme-Modified Carbon Nanotube Columns', Chem. Commun., 2017, 53, 9839-9841; (2) Reeve, H.A., Ash, P.A., Park, H., Huang, A., Posidias, M., Tomlinson, C., Lenz, O., Vincent, K.A., 'Enzymes as modular catalysts for redox half reactions in H2-powered chemical synthesis: from biology to technology', Biochemical Journal, 2017, 474, 215-230; (3) Reeve, H.A., Lauterbach, L., Lenz, O., Vincent, K.A. 'Enzyme-Modified Particles for Selective Bio-Catalytic Hydrogenation via H2-driven NADH Recycling' ChemCatChem, 2015, 7, 21, 3480-3487.
Start Year 2014
 
Title COFACTOR REGENERATION SYSTEM 
Description European Patent Granted 5 December 2018 to protect a cofactor recycling system, using electrically conducting supports modified with enzymes for hydrogen-driven NADH recycling. 
IP Reference EP2764107 
Protection Patent granted
Year Protection Granted 2014
Licensed No
Impact Commercial negotiations in progress.
 
Description 'A collaborative journey to greener chemicals' - Article for Adjacent Government 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Wrote an article about the impact that RCUK funding has had on our research and the potential impact of our research. Article highlights the benefit of funding for collaborations with Industry and the impact this can have on development of innovative technologies, specifically in the biotech sector.
Year(s) Of Engagement Activity 2016
URL http://www.adjacentopenaccess.org/farming-environment-marine-sustainable-news/collaborative-journey-...
 
Description 'Award-winning HydRegen technology offers path to 'clean, safe' chemical production' - Article on University's Science Blog 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact The University of Oxford's media team wrote an article about the HydRegen Technology to promote the research to a wide audience.
Year(s) Of Engagement Activity 2016
URL http://www.ox.ac.uk/news/science-blog/award-winning-hydregen-technology-offers-path-clean-safe-chemi...
 
Description 'Cleaner greener method for making lab chemicals' - Radio Show (The Science Show on Australian Broadcasting Commission) 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Kylie Vincent and Holly Reeve were interviewed by Robyn Williams on the ABC radio show - The Science Show. This show has a large audience in Australia and world wide.The purpose of the interview was to explain the HydRegen technology to a general, but scientifically interested, audience. This has led to an increase in awareness of our work, biocatalysis and industrial biotechnology internationally.
Year(s) Of Engagement Activity 2016
URL http://www.abc.net.au/radionational/programs/scienceshow/cleaner-greener-method-for-making-lab-chemi...
 
Description 'Learning from nature' - podcast on the University of Oxford's 'Big Questions' series 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact A podcast about how bacteria may hold the key to using hydrogen as a clean energy source, both as a fuel and as a route to chemicals.
Year(s) Of Engagement Activity 2016
URL http://www.oxfordsparks.ox.ac.uk/content/learning-3-learning-nature
 
Description 2018 STEM Christmas Lecture at Natural History Museum, Oxford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact PDRA on the project, Jack Rowbotham, gave the Annual STEM Christmas Lecture at the Natural History Museum, Oxford. Around 100 secondary school students attended.
Year(s) Of Engagement Activity 2018
 
Description Animation and teaching resources 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Schools
Results and Impact Created an animation with the University public engagement team. The animation highlights the importance of industrial biotechnology as well as the technology developed as part of this project. Resources for schools were also produced. This animation was put on social media by the University of Oxford and reached more than 10,000 views in the first month.
Year(s) Of Engagement Activity 2017
URL http://www.oxfordsparks.ox.ac.uk/content/what-can-chemists-learn-nature
 
Description Annual Women in Science event at Jesus College, Oxford 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Lecture to inspire female 6th form school students about applying to STEM subjects at University, presented by project PDRA, Lisa Thompson.
Year(s) Of Engagement Activity 2019
 
Description Exhibit at the University of Oxford's European Researchers' Nigh event - Curiosity Carnival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact We held an exhibit at the Curiosity Carnival in the Natural History Museum (Oxford). More than 2000 people attending during the evening. Our exhibit - What can Chemists Learn from Nature? centered on bio-inspired technologies. We had a game where people could match technologies to the bit of nature that inspired it, we showcased the animation we have produced about the area of industrial biotechnology and we performed live experiments (#RacingEnzymes) to show how nature can be intensified in the lab to make the chemicals we need. The event sparked questions and discussions in the area of industrial biotechnology and inspired students about the meeting point of the sciences as well as the overlap between academia and industry.
Year(s) Of Engagement Activity 2017
URL http://www.ox.ac.uk/curiosity-carnival/about
 
Description STEM Christmas lecture for 300 year 10 students 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Gave a presentation called: Inspired by nature: from planes and trains, to your box of Christmas chocolates! 300 students attended.
Year(s) Of Engagement Activity 2017
 
Description School outreach demonstration and lecture 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Research-related demonstrations and lectures given to 5 secondary classes at Hodge Hill Girls School, Birmingham, to inspire female students to consider careers in STEM and educate them about research in biotechnology.
Year(s) Of Engagement Activity 2018
 
Description Smarter Smells - Interview for a Chemistry World article 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Interviewed for a Chemistry World articles abut biocataylsis for the production of flavour and fragrances.
Year(s) Of Engagement Activity 2016
URL https://www.chemistryworld.com/feature/smarter-smells/1017487.article
 
Description YouTube channel to promote biotechnology research 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Series of podcasts and animations educating the public about biotechnology
Year(s) Of Engagement Activity 2018
URL https://www.youtube.com/channel/UCq9YqdQt4oLtXZ1Y94a7-uw