Innovative Routes to Monoterpene Hydrocarbons and Their High Value Derivatives

Propelling chemicals/natural products production towards 'green' and more sustainable manufacturing processes will require the harnessing of the power of Synthetic Biology (SynBio) and integration with more traditional biocatalytic and chemo-catalytic processes. SynBio is a potentially disruptive technology, but major de-risking is required before the SynBio of chemicals manufacture can replace traditional manufacturing processes. In part, this is reflected in the major costs associated with the development of new manufacturing plant in the industrial sector, which is required to transition from conventional petrochemical/natural resource-based manufacturing to SynBio-based chemicals manufacturing.

In this programme we will design, build and integrate new SynBio technology that accelerates delivery of bespoke SynBio solutions for chemicals/natural products synthesis. Our focus is on a major group of industrially valuable compounds, the monoterpenoids, which offer new opportunities for bio-based production using SynBio. By adopting modular 'plug-and-play' platform approaches and a production pipeline that embraces the 'design-build-test-deploy' life-cycle we will turn knowledge assets into innovative chemicals production solutions for monoterpenoid compounds. We will recruit and design bespoke biological parts (e.g. enzyme catalysts; transporters; membrane components) and assemble them in novel ways to create a bio-based production pipeline contained within a synthetic, engineered microbial biofactory. Iterative design-build-test-deploy life-cycles will be used to optimise production of monoterpenoid chemicals in defined proof-of-principle projects. Using these Demonstrator Projects, we will deliver innovative SynBio approaches for chemicals/natural products biosynthesis and promote leading capability development that will stimulate interactions with industry and other stakeholders. These projects will require the unique, multidisciplinary environment (including bioscience, engineering, chemistry and computational science) and collaborative working culture represented by the team of applicants.

Enzyme-catalysed SynBio-based synthesis is attractive: regio- and stereo-selective biotransformation, use of 'green' reaction conditions and lack of toxic by-products combine to make biosynthesis attractive in sustainable manufacturing. US and European legislation acknowledges that products from biosynthetic manufacturing processes are considered 'natural'. This adds significant market value, especially in foods and flavourings manufacture, as is the case in the production and use of monoterpenoids. The SynBio approaches we develop will also generate new chemical entities not readily accessible using traditional synthetic approaches. This will provide new opportunities for industrial exploitation, including the synthesis of new chemical libraries that will support industrial and academic drug discovery programmes. More broadly, the programme will provide general tools, technology platforms and SynBio 'know-how' that will impact widely in sustainable manufacture of chemicals and natural products and de-risk SynBio approaches for more rapid take-up and development by the industrial sector.

Terpene cyclases/synthases (TC/S) catalyse the most complex reactions in biology and are the entry points to the most structurally and stereochemically diverse group of natural products, the terpenoids. Despite its rich chemical diversity, the terpenome is derived from common metabolites through a series of complex reactions initiated by the TC/S enzyme family. This application is focussed on cyclic/acyclic monoterpene hydrocarbons and their high value derivatives (monoterpenoids). These compounds are found in plant extracts and essential oils, and can be derived from petrochemical feedstocks. They are synthesised through the activity of the large TC/S family, which catalyse 'high energy' cyclisation reactions involving unstable carbocation intermediates.

Through plug-and-play engineering we will link knowledge of TC/S chemistry with metabolic engineering to create novel biosynthetic pathways for important monoterpenoids. We will generate tools and knowledge to develop robust microbial strains for monoterpenoid biosynthesis. We will integrate expertise across several knowledge areas including: enzyme/pathway building block discovery; design and engineering for creation of a library of parts; refactoring of novel metabolic pathways; genome-scale metabolic modelling and experimental pathway validation; chassis engineering and end-product yield optimisation. A feature of the programme is the need to integrate knowledge of enzyme mechanism with bio/chemo-catalysis, systems biology (SysBio) and SynBio, including pathway/chassis engineering.

Our vision is to propel the production of monoterpenoids towards 'green' and sustainable manufacturing by harnessing the power of SynBio and linking this to detailed understanding of TC/S chemistry. We will provide essential tools, resources and basic science 'know-how' that de-risks the application of SynBio to monterpenoid production and open up SynBio routes to synthesis of the chemically diverse monoterpenome.

Global trends are driving an increasing demand for bio-based chemical production. With the historic volatility of global oil prices, limited and seasonal variation in natural resources, global pollution, consumers becoming eco-aware and desiring to live more sustainably, governments and companies are acting to implement policies/processes to reduce global dependence on existing natural/oil-based products and transition to more sustainable ways of living. This programme is at the heart of this agenda developing bio-based SynBio technology for chemicals manufacture of commercially important monoterpenoids.
Benefits from the research:
(i) People. We will train a new generation of scientists at all levels who can work across discipline boundaries, harnessing expertise in predictive design and the SynBio of chemicals biosynthesis. This will capture innovative approaches being developed in chemo/bio-catalysis process design, SynBio, experimental and computational SysBio, and knowledge-based biocatalyst/component design fields. An ability to integrate skills from these different research areas to develop novel bio-catalytic solutions for industry will be a major part of training. We will enable training and delivery of project milestones in laboratory environments built to facilitate working across biological and chemical disciplines, supported by strengths in computation, biophysics and mechanistic chemistry, structural biology, SynBio and SysBio. Training in platform technologies and Responsible Innovation will produce staff who will become SynBio ambassadors.
(ii) Knowledge. Bio-based chemical synthesis through IB and SynBio has the potential to offer powerful solutions to major societal challenges such as health, energy supply and sustainable manufacture. This will be driven through integration of biological and chemical processes and major changes to chemical production for agriculture, pharmaceuticals, and bulk chemicals manufacturing across a range of sectors. These are burgeoning economies, restricted in scope through our ability to design/re-profile enzyme catalysts, requiring innovative chemo-/bio-catalytic solutions to processing and the coupling of chemo- and bio-catalysis through SynBio approaches to facilitate novel chemical transformations. These new developments will drive wealth creation in chemicals manufacturing and its allied economies. There is strong potential for inward investment from industrial stakeholders, and the possibility of technology licensing/spin out/spin in opportunities for technologies developed in the research programme.
(iii) Economy. The programme is positioned predominantly in the TRLs 1-3 space, and our mission will be to accelerate commercialisation with our partners in a 'market-focused approach' by driving the technology we develop towards the higher TRLs. We have identified goals and strategies to achieve these aims and a number of mechanisms for attaining targets/deliverables. Early IP targets will be focused on new tools for SynBio, including development of novel building blocks/components for redesign, and the engineering of novel biocatalysts, regulatory elements and chassis. Industry will benefit from these tools. Novel processes for chemicals/natural products synthesis based on monoterpene scaffolds will emerge, which will foster economic competitiveness in the UK in the area of chemicals manufacture. The work will impact quality of life by adopting sustainable, green processes for manufacture and provide tools/resources more broadly for the chemicals manufacturing industry over the 5 years of the award.
The Responsible Innovation elements of our programme will include real-time assessment and anticipation of research and innovation trajectories, deliberation and reflection, and collaborative development. This will impact by informing publics, policy-makers and wider stakeholders about the impact and benefits/limitations of SynBio in chemicals manufacture.