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

来自不同生物领域的碳水化合物活性酶的发现、表征和应用

• 批准号: RGPIN-2018-03892
• 负责人: Brumer, Harry
• 金额: $ 7.14万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=692605
• 关键词: Discovery; characterization; application; carbohydrate; active

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.

复杂碳水化合物的酶促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转运蛋白的特异性对于生物学至关重要。 基础酶的发现对于开发高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对植物和环境微生物（特别是瘤胃细菌、腐生细菌和腐生/植物病原真菌）中的单个酶和酶群的功能提供新的见解。 将使用先进的分子系统发育方法来挖掘基因组和转录组，以识别目标 CAZymes、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，以及通过完善的合作进行结构生物学和反向遗传分析。 该项目以其广度和基于系统的方法在加拿大独一无二，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的发展：(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) 对环境细菌的复杂多糖利用位点进行系统分析 (3) 发现新的 用于（生物）材料功能化的铜自由基氧化酶。 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以促进新型酶和微生物技术向工业转移方面具有巨大潜力。 通过多学科交叉培训，包括技术、沟通、团队合作和指导技能，该计划将让 HQP 成为加拿大高科技林业、农业、酶技术和一系列其他生物技术行业的领导角色。