FPLC System for Bioactive Peptides Research

用于生物活性肽研究的 FPLC 系统

• 批准号: RTI-2021-00371
• 负责人: Aluko, Rotimi
• 金额: $ 8.15万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=718583
• 关键词: FPLC; System; Bioactive; Peptides; Research

Isolation, purification, functional characterization, and in vivo testing of bioactive peptides require qualitative and quantitative analysis using the fast protein liquid chromatography (FPLC) system. At the Department of Food and Human Nutritional Sciences, University of Manitoba, there is only one FPLC system, which was purchased in 2004 but is no longer serviceable because the model has been discontinued and there is no suitable FPLC system on the University of Manitoba campuses. Therefore, the aim of this proposal is to replace the non-serviceable 16-year old system with a new FPLC system that will provide critical tools for continued operation of my NSERC-funded Discovery research program on bioactive peptides. The FPLC provides the most rapid and cost-effective means of separating and isolating peptides in forms that can be characterized for structure-function properties or further purified on a higher pressure system (HPLC). Moreover, the FPLC facilitates rapid quantitative estimation of peptides of defined molecular sizes, which is a critical component of structural elucidation and a major goal of my NSERC-funded Discovery research program. The research program focuses on elucidation of the structure-function properties of food protein-derived bioactive peptides, which are usually contained in enzymatic digests of food proteins called protein hydrolysates. Because enzymatic hydrolysates contain a mixture (sometimes hundreds) of peptides, the initial rate-determining step for successful structure-function analysis is separation on an FPLC system. Thus, the FPLC is a required component of my Discovery research work and for successful training of highly qualified personnel (HQP). The FPLC will be used to separate bioactive peptides into fractions that are suitable for structural elucidation (amino acid sequencing) by liquid chromatography-tandem mass spectrometry. Knowledge of the amino acid sequence is required for the discovery of highly potent peptides that can serve as effective modulators of various metabolic pathways. Thus the FPLC system is a valuable tool in my quest to provide fundamental scientific information on the mechanism of action of bioactive peptides as well as the discovery of novel peptides that can reduce or prevent negative metabolic events. The system will also facilitate my research that seeks discovery of highly functional food ingredients that can enhance nutritional status of Canadians while promoting value-added utilization of Canadian crops and animal products. This is because results obtained from the instrument will provide researchers with the ability to design novel food ingredients that can be used to formulate functional foods and nutraceuticals. The FPLC system will be used by graduate and undergraduate students, postdoctoral fellows/research associates, and technicians on a daily basis for peptide fractionation and quantification. Therefore, the instrument is critical to the successful training of HQPs.

生物活性肽的分离、纯化、功能表征和体内测试需要使用快速蛋白质液相色谱 (FPLC) 系统进行定性和定量分析。曼尼托巴大学食品与人类营养科学系只有一套FPLC系统，该系统于2004年购买，但由于该型号已停产且曼尼托巴大学校园内没有合适的FPLC系统而不再使用。因此，本提案的目的是用新的 FPLC 系统替换已经无法使用的 16 年旧系统，该系统将为我的 NSERC 资助的生物活性肽发现研究计划的持续运行提供关键工具。 FPLC 提供了最快速、最具成本效益的分离肽的方法，其形式可以表征结构功能特性或在高压系统 (HPLC) 上进一步纯化。此外，FPLC 有助于快速定量估计特定分子大小的肽，这是结构阐明的关键组成部分，也是我的 NSERC 资助的发现研究计划的主要目标。该研究计划的重点是阐明食品蛋白质衍生的生物活性肽的结构功能特性，这些生物活性肽通常包含在称为蛋白质水解物的食品蛋白质的酶消化物中。由于酶水解产物含有多种肽的混合物（有时数百种），因此成功进行结构功能分析的初始速率确定步骤是在 FPLC 系统上进行分离。因此，FPLC 是我的 Discovery 研究工作和成功培训高素质人员 (HQP) 所必需的组成部分。 FPLC 将用于将生物活性肽分离成适合通过液相色谱-串联质谱法进行结构阐明（氨基酸测序）的级分。为了发现可以作为各种代谢途径的有效调节剂的高效肽，需要了解氨基酸序列。因此，FPLC 系统是我寻求提供有关生物活性肽作用机制的基本科学信息以及发现可以减少或预防负面代谢事件的新型肽的宝贵工具。该系统还将促进我的研究，寻求发现高功能食品成分，可以提高加拿大人的营养状况，同时促进加拿大农作物和动物产品的增值利用。这是因为从该仪器获得的结果将为研究人员提供设计新颖食品成分的能力，这些食品成分可用于配制功能性食品和营养保健品。 FPLC 系统将供研究生和本科生、博士后研究员/研究员和技术人员日常使用，以进行肽分级和定量。因此，该仪器对于指挥员的成功培训至关重要。