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

复杂碳水化合物的酶合成和降解是自然界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，以及相关碳水化合物结合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。基础酶的发现对于开发有效的工业生物技术应用同样至关重要，从食品、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。在这种背景下，我的研究计划的中心愿景是为植物和环境微生物（特别是瘤胃细菌，腐生细菌和腐生/植物致病真菌）的单个酶和酶联合体的功能提供新的见解。基因组和转录组将使用先进的分子系统发育方法进行挖掘，以确定目标酶、碳水化合物结合蛋白和tonb依赖性转运蛋白，用于重组生产和详细的酶学/生化表征，以及结构生物学和反向遗传分析。该项目在加拿大独一无二，其广度和基于系统的方法将通过三个主要目标推进碳水化合物酶学和糖生物学领域：(1)阐明植物细胞壁生物学中GH16 β -葡聚糖水解酶和转糖基酶的功能进化；(2)环境细菌中复杂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。该计划的重点将是产生高影响力的基础研究成果，尽管在产生知识产权方面有很大的潜力，从而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，该计划将使HQP在加拿大的高科技林业、农业、酶技术和一系列其他生物技术产业中发挥主导作用。

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.