Biocatalytic Activation of Macromolecular Building Blocks

更多地利用所有生物资源部分将对一个在加拿大不断增长的生物经济和减少碳排放的国家承诺中发挥核心作用的森林部门至关重要。此外，抓住这一机会将实现加拿大供应全球生物产品市场的潜力，同时利用其在生物资源使用和管理方面的基础设施和专门知识的历史投资。 除了可再生生物质原料的创新使用外，生物经济还通过增加可持续的生物催化工艺的应用来体现。事实上，微生物酶对扩大可由植物来源的生物质制成的产品范围的重要性是众所周知的。然而，迄今为止的生物技术主要侧重于解构生物资源，用于微生物发酵或化学转化为商品燃料和化学品。虽然衍生自初级糖和单木质醇的产品对于捕获可再生植物生物质的全部潜力是必要的，但这些产品放弃了起始生物质资源的固有功能属性。 生物催化剂升级（而不是降解）生物聚合物化学的潜力提出了一种替代的做法，“解构重建”，占主导地位的当前生物精炼概念。因此，而不是目标酶的木质纤维素解构，拟议的计划将集中在酶系统的发现和应用，激活未充分利用的半纤维素片段，允许它们重新组装成生物聚合物结构，捕获这个主要的生物质部分的固有功能，导致生物基交联剂，表面涂层和流变改性剂。对生物催化途径的重视将有助于确保最终产品和合成途径都是可持续的，同时也利用了生物催化方法提供的催化精度。为实现该计划的目标，将采取以下四个综合步骤：1）生物信息学分析，优先选择碳水化合物氧化酶和转氨酶，并为蛋白质工程策略提供信息; 2）使用高通量和重组方法生产蛋白质靶点; 3）分离的酶和反应产物的生物化学表征以鉴定互补和相容的酶系统，和4）应用已建立的偶联途径来证明重新组装酶促活化的寡聚体的潜力。除了加速基因组学研究在生物制品领域的应用外，拟议的研究还将推动无细胞生物催化级联的新兴趋势，并创造新的工具，以促进从未充分利用的生物质馏分中设计和开发新型生物基材料。

Greater use of all bioresource fractions will be critical to an invigorated forest sector that plays a central role in Canada's growing bioeconomy and national commitments to reduced carbon emissions. Moreover, seizing this opportunity will realize Canada's potential to supply global bioproduct markets while capitalizing on its historical investment in infrastructure and know-how in bioresource use and management. In addition to innovative use of renewable biomass feedstocks, bio-economies are exemplified by increased application of sustainable, biologically catalyzed processes. Indeed, the importance of microbial enzymes to expanding the range of products that can be made from plant-derived biomass is well recognized. However, biotechnologies to date largely focus on the deconstruction of the bioresource for microbial fermentation or chemical conversion to commodity fuels and chemicals. While products derived from primary sugars and monolignols will be necessary for capturing the full potential of renewable plant biomass, these products relinquish the inherent functional attributes to the starting biomass resource. The potential of biocatalysts to upgrade (rather than degrade) biopolymer chemistries presents an alternative to the practice of “deconstruct to reconstruct” which dominates current biorefinery concepts. Accordingly, rather than target enzymes for lignocellulose deconstruction, the proposed program will focus on the discovery and application of enzyme systems that activate underused hemicellulose fragments, permitting their reassembly into biopolymer structures that capture the innate function of this major biomass fraction, leading to bio-based crosslinkers, surface coatings, and rheology modifiers. The emphasis on biocatalytic routes to activated building blocks will help ensure that both end-products and synthesis pathways are sustainable, while also harnessing the catalytic precision offered by a biocatalytic approach. The following four integrated steps will be followed to achieve the program goals: 1) bioinformatics analyses to prioritize carbohydrate oxidase and transaminase selections and inform protein engineering strategies; 2) production of protein targets using high-throughput and recombinant approaches; 3) biochemical characterization of isolated enzymes and reaction products to identify complementary and compatible enzyme systems, and 4) application of established coupling pathways to demonstrate the potential to re-assemble the enzymatically activated oligomers. In addition to accelerating the application of genomics research in the bioproducts sector, the proposed research will advance emerging trends in cell-free biocatalytic cascades, and create new tools to facilitate the design and development of novel bio-based materials from underused biomass fractions.