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

不断增加的全球能源消耗正在加速化石燃料的枯竭。当石油储量最终耗尽时，提炼过程中产生的重要副产品将不再可用。由于这些芳香化学物质用于许多过程，包括塑料、洗涤剂、化肥、药品、油漆和合成纤维的制造，因此找到替代来源至关重要。木质素是植物的“木质”部分，因此它可以可持续地生产，它是可再生的，是一种天然的碳储存形式。重要的是，它是一种由芳香族化学成分组成的复杂聚合物，是上述不可再生芳香族炼油副产品的潜在替代品。我们的目标是测试从木质素中生产低分子量芳香化学原料的可行性，木质素目前是木材加工和造纸的废物，因此它可能被用来制造有用的产品。我们建议开发一个“前端”来优化木质素转化为其组成的芳香化学构件。这项技术可以被固定在生物精炼厂的任何“后端”，以生产生物塑料、生物表面活性剂、生物材料等等。通过探索和优化一种允许细菌或真菌快速调整以利用木质素转化的技术，我们能够通过优化用作化学原料的降解产物和使最终生物产品的范围多样化来限制浪费。提出的工作使用一种新的合成生物学平台来创建基因组合，从而使废木质素的最佳降解（关于效率和产生的产品）。然后，这些组合可以用作在迭代设计-制造-测试过程中生成进一步组合的起点。这将在提高木质素转化为可用原料的产量方面为用户创造价值。除了优化任何特定应用的酶混合物外，对结果的分析将使团队能够开发启发式方法，这将有助于未来整个生物质处理系统的合理设计，并将导致对生物质降解过程的更深入了解。因此，简而言之，我们建议使用合成生物学来设计能够酶解造纸工业废物的微生物，以提供芳香原料分子的新来源，确保这些分子向工业供应，并确保社会可以继续使用我们都认为理所当然的日常物品。该项目的一个完整阶段将涉及应用最先进的分析技术，积极识别离散的低分子量芳香化学品，以产生来自新型工程木质素分解活性的可行商业线索。开发的技术将由Ingenza有限公司提供给市场。有许多可能的商业化途径，包括：1)生物制造大量的酶混合物，作为细胞裂解物或提取物直接销售给生物精炼厂运营商。二)。或者，特定细胞系本身可以在许可下提供给客户，作为木质素分解加工工具。iii)。为希望为他们感兴趣的特定生物质实施定制消化底盘的客户提供合同服务；可能需要某种形式的应变维护和进一步的修改。

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.