Collaborative Research: Expanding the substrate range of Clostridium thermocellum to improve understanding of its metabolism

合作研究：扩大热纤梭菌的底物范围，以增进对其代谢的了解

• 批准号: 2313152
• 负责人: Daniel Olson
• 金额: $ 47.99万
• 依托单位: Dartmouth College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2313152&HistoricalAwards=false
• 关键词: Collaborative; Research; Expanding; substrate; range

The dry matter in plants is called lignocellulosic biomass. It is the most abundant source of renewable raw material in the world. Converting it to fuels and products would contribute substantially to a circular bioeconomy. Unfortunately, parts of it are difficult to deconstruct, which increases costs and lowers yields. Certain microbes are capable of deconstructing lignocellulose. The objective of this project is to elucidate the metabolism of one such microbe. Once understood, efforts will be made to expand the range of substrates the organism can utilize. In parallel with the research activities, both primary investigators will work through established channels to engage K-12 students in science fairs and summer research experiences. The limited substrate range of natively cellulolytic organisms (such as C. thermocellum) makes it difficult to understand their metabolism and engineer them for applied purposes. The goal of this study is to expand the substrate range of C. thermocellum to improve our understanding of its metabolism to diagnose limitations to product formation. To accomplish this, the following set of coordinated aims will be pursued. (1) Characterize substrate transport and reversibility of central metabolism to determine factors that limit substrate range. (2) Identify mutations that allow growth on alternative substrates, using a combination of stationary-phase evolution and a combinatorial multi-substrate chemostat-based approach. (3) Use alternative growth substrates to determine factors that limit ethanol production. The work in this proposal will improve our understanding of atypical forms of glycolysis, which will expand our understanding of the possible variations of this ubiquitous and highly conserved metabolic pathway. It will improve our understanding of the factors that control substrate range in organisms, including limitations due to transport and metabolic reaction reversibility. It will also allow us to understand factors that limit product titer, a central challenge in the field of metabolic engineering.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

植物中的干物质称为木质纤维素生物质。它是世界上最丰富的可再生原材料来源。将其转化为燃料和产品将为循环生物经济做出重大贡献。不幸的是，它的某些部分很难解构，这增加了成本并降低了产量。某些微生物能够解构木质纤维素。该项目的目标是阐明一种此类微生物的新陈代谢。一旦理解，将努力扩大生物体可以利用的底物范围。在开展研究活动的同时，两位主要研究者将通过既定渠道让 K-12 学生参与科学博览会和夏季研究体验。天然纤维素分解生物（例如热纤梭菌）的底物范围有限，因此很难了解它们的代谢并对其进行改造以用于应用目的。本研究的目的是扩大热纤梭菌的底物范围，以提高我们对其代谢的理解，从而诊断产物形成的局限性。为了实现这一目标，将追求以下一系列协调一致的目标。 (1) 表征底物转运和中心代谢的可逆性，以确定限制底物范围的因素。 (2) 结合使用稳定期进化和基于组合多基质恒化器的方法，识别允许在替代基质上生长的突变。 (3) 使用替代生长底物来确定限制乙醇生产的因素。该提案中的工作将增进我们对糖酵解非典型形式的理解，这将扩大我们对这种普遍存在且高度保守的代谢途径的可能变化的理解。它将提高我们对控制生物体底物范围的因素的理解，包括运输和代谢反应可逆性造成的限制。它还将使我们能够了解限制产物滴度的因素，这是代谢工程领域的一个核心挑战。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。