Development of oxidases for synthesis of bioplastics intermediates

开发用于合成生物塑料中间体的氧化酶

• 批准号: BB/V003100/1
• 负责人: Andrew Carnell
• 金额: $ 32.19万
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV003100%2F1
• 关键词: Development; oxidases; synthesis; bioplastics; intermediates

Due to environmental pressure, there is an urgent need to develop novel, high-performance, biodegradable and sustainable plastics from bio-renewable feedstocks. Plant lignocellulose-derived 2,5-furan dicarboxylic acid (FDCA) has great potential for replacing petroleum-derived chemicals in various plastic polymers, for instance in polyethylene terephthalate (PET). Bio-derived and biodegradable plastics incorporating FDCA, such as poly(ethylene furanoate) (PEF) and poly(butylene adipate-co-butylene furandicarboxylate) (PBAF) are being developed. However, currently there are no large-scale producers of the FDCA monomer which is needed to support large scale production of these potentially high-value polymers. This project addresses the need to a develop novel process for the production of FDCA from biomass which will feed into bioplastic manufacturing pipelines for which there is growing demand.Enzymatic conversions confer advantages over traditional industrial chemical routes as they require less energy and are carried out in milder conditions. Enzymes often permit highly selective production of desired target compounds and deliver a higher level of purity. This is particularly important when producing monomers such as FDCA since impurities can interfere with the polymerisation process. Recent research at Liverpool in collaboration with Biome Bioplastics and Leeds University showed proof-of-concept for a process that uses a combination of 4 enzymes to produce FDCA, some of which were difficult to produce on scale. At Liverpool we identified a single new enzyme (E56) which could replace 3 of these enzymes. While the enzyme is unique in this ability, its efficiency is currently low. We also discovered another new enzyme (E5), with higher activity, but that works for part of the pathway and could be used as a combination with the other new enzyme. This project seeks to employ advanced synthetic biology techniques and state-of-the art equipment to improve the enzymes' capabilities through directed evolution. This technique allows evolution of enzyme catalytic activity in the laboratory and the approach will involve generating and screening a very large number of genetic variants. Having identified new enzymes, we will test their activity and suitability as a drop-in replacement for the previously used enzymes.The project outcomes will include development of improved enzymes that can be used in the production of bioplastic precursors, and establishment of an efficient ultra-high throughput screening platform that can be applied to other enzymes in need of improvement.

由于环境压力，迫切需要以生物可再生原料为原料开发新型、高性能、可生物降解和可持续发展的塑料。植物木质纤维素衍生的2，5-呋喃二酸(FDCA)在各种塑料聚合物中取代石油衍生的化学物质具有很大的潜力，例如在聚对苯二甲酸乙二醇酯(PET)中。含有FDCA的生物衍生和生物可降解塑料，如聚(呋喃甲酸乙二醇酯)(PEF)和聚(己二酸丁二醇酯-呋喃甲酸丁二醇酯)(PBAF)正在开发中。然而，目前还没有大规模的FDCA单体生产商，这是支持这些潜在高价值聚合物大规模生产所需的。该项目致力于开发一种从生物质中生产FDCA的新工艺，这些生物质将被输送到需求不断增长的生物塑料制造管道中。与传统的工业化学路线相比，酶转化具有优势，因为它们需要更少的能源，并且在较温和的条件下进行。酶通常允许高度选择性地生产所需的目标化合物，并提供更高水平的纯度。这在生产FDCA等单体时尤其重要，因为杂质会干扰聚合过程。利物浦最近与Biome BioPlatform和利兹大学合作进行的研究表明，使用4种酶组合生产FDCA的工艺是概念验证的，其中一些很难规模化生产。在利物浦，我们确定了一种新的酶(E56)，它可以取代其中的3种酶。虽然这种酶在这种能力上是独一无二的，但它的效率目前很低。我们还发现了另一种新的酶(E5)，活性更高，但它对该途径的一部分起作用，可以与另一种新的酶结合使用。该项目寻求使用先进的合成生物学技术和最先进的设备，通过定向进化来提高酶的能力。这项技术允许在实验室中进化酶的催化活性，该方法将涉及产生和筛选非常大量的遗传变异。在找出新的酵素后，我们会测试它们的活性和适合性，以替代以往使用的酵素。计划的成果包括开发可用于生产生物塑胶前体的改良酵素，以及建立一个有效的超高通量筛选平台，可应用于其他需要改良的酵素。