Discovery, characterization, and application of carbohydrate-active enzymes from diverse biological niches

酶促合成和降解复合碳水化合物是自然界中最重要的生物催化过程之一。因此，阐明碳水化合物活性酶(CAZymes)的催化多样性以及相关的碳水化合物结合蛋白和碳水化合物运输蛋白的特异性对生物学具有至关重要的基础意义。基础酶的发现同样对开发高效的工业生物技术应用至关重要，从用于食品、饲料、燃料和材料的生物质加工，到有机生物催化和医疗用途。在此背景下，我的研究计划的中心愿景是对来自植物和环境微生物(特别是瘤胃细菌、腐生细菌和腐生/植物病原真菌)的单个酶和酶联合体的功能提供新的见解。将使用先进的分子系统学方法挖掘基因组和转录本，以确定目标CAZymes、碳水化合物结合蛋白和TonB依赖的转运体，用于重组生产和详细的酶/生化特征，以及结构生物学和反向遗传分析。该计划在加拿大的广度和系统方法方面是独一无二的，它将通过三个主要目标推动碳水化合物酶学和糖生物学领域的发展：(1)阐明GH16β-葡聚糖水解酶和转糖基酶在植物细胞壁生物学中的功能进化；(2)环境细菌复杂的多糖利用位点的系统分析；(3)发现用于(生物)材料功能化的新型铜自由基氧化酶。该计划的重点将是生产高影响力的基础研究成果，尽管知识产权的产生有巨大的潜力，可以将新的酶和微生物技术转移到工业中。通过多学科交叉培训，包括技术、沟通、团队合作和指导技能，该计划将在HQP阶段进入加拿大高科技林业、农业、酶技术和一系列其他生物技术行业的领先地位。

The enzymatic synthesis and degradation of complex carbohydrates ranks among the most important biocatalytic processes in Nature. Thus, illuminating the catalytic diversity of carbohydrate-active enzymes (CAZymes), as well as the specificity of associated carbohydrate-binding and carbohydrate-transport proteins is of crucial fundamental importance to biology. Fundamental enzyme discovery is likewise essential to the development of efficient industrial biotechnology applications, ranging from biomass processing for food, feed, fuel, and materials, to organic biocatalysis and medical uses. In this context, the central vision of my research program is to provide new insight into the function of individual enzymes and enzyme consortia from plants and environmental microorganisms (specifically ruminal bacteria, saprophytic bacteria, and saprophytic/phytopathogenic fungi). Genomes and transcriptomes will be mined using advanced molecular phylogenetic methods to identify target CAZymes, carbohydrate-binding proteins, and TonB-dependent transporters for recombinant production and detailed enzymological/biochemical characterization, as well as structural biology and reverse-genetic analysis via well-established collaborations. Unique in Canada in terms of its breadth and systems-based approach, this program will advance the fields of carbohydrate enzymology and glycobiology through three primary Objectives: (1) Elucidating the functional evolution of GH16 beta-glucan hydrolases and transglycosylases in plant cell wall biology, (2) Systems analysis of complex polysaccharide utilization loci from environmental bacteria (3) Discovery of novel copper-radical oxidases for (bio)materials functionalization. The focus of this program will be the production of high-impact, fundamental research output, although there is significant potential for the generation of intellectual property leading to the transfer of novel enzyme and microbial technologies to industry. Through multidisciplinary cross-training, including technical, communication, teamworking, and mentorship skills, the program will stage HQP to enter leading roles in Canada's high-tech forestry, agricultural, enzyme technology, and a range of other biotechnology industries.