13TSB_SynBio Ligniflex: A synthetic biology platform to optimise the process and products of enzymatic lignin disruption

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Increasing global energy consumption is accelerating the rate of fossil fuel depletion. When oil reserves are eventually exhausted the essential by-products produced in refining will no longer be available. Since these aromatic chemicals are used in many processes including the manufacture of plastics, detergents, fertilizers, pharmaceuticals, paints and synthetic fibres it is vital that alternative sources become available.Lignin is the 'woody' part of plants and therefore it can be produced sustainably, it is renewable and a natural form of carbon storage. Importantly for this proposal, it is a complex polymer made of aromatic chemical building blocks, potential replacements for the aforementioned non-renewable aromatic oil-refining by-products.Our goal is to test the feasibility of producing low molecular weight aromatic chemical feedstocks from the lignin that is currently a waste product from wood processing and paper manufacturing, so that it may be used to manufacture useful products. We propose to develop a "front-end" to optimise the conversion of lignin into its constitutive aromatic chemical building blocks. This technology may be bolted to any "back-end" in a biorefinery to produce bioplastics, biosurfactants, biomaterials and so on. By exploring and optimising a technology which allows for the rapid tuning of bacteria or fungi for exploiting the conversion of lignin, we stand to limit waste by being able to optimise the degradation products being used as chemical feedstocks and diversify the range of end-bioproducts possible.The proposed work uses a novel synthetic biology platform to create gene combinations which give the optimal degradation (with regards to efficiency and products produced) of waste lignin. These combinations may then be used as a starting point to generate further combinations in an iterative design-make-test process. This will create value for the user in terms of improvements in the yield of lignin conversion to useable feedstock. In addition to enabling the optimisation of enzyme blends for any given application, analysis of the results will allow the team to develop heuristics which will facilitate the rational design of whole biomass processing systems in the future, and will lead to a deeper understanding of biomass degradation processes.Therefore, in brief, we are proposing to use synthetic biology to engineer microorganisms capable of enzymatically degrading waste from the paper manufacturing industry to provide a new source of aromatic feedstock molecules, securing the supply of these molecules to industry and ensuring society can continue to use the everyday items we all take for granted. An integral phase of the project will involve applying state of the art analytical techniques to positively identify discrete low molecular weight aromatic chemicals to produce viable commercial leads derived from novel engineered lignolytic activity.The technology developed will be offered to the marketplace by Ingenza Ltd. There are a number of possible routes for commercialisation including: i.) Biomanufacture of bulk quantities of the enzyme blend for sale direct to biorefinery operators as cell lysate or extract. ii.) Alternatively, the specific cell line itself could be made available under license to a customer, as a lignolytic processing tool. iii.) Provision of a contract service to customers wishing to have a bespoke digestion chassis implemented for the particular biomass of interest to them; probably involving some form of strain maintenance and further modification as required.