Metabolic adaptability of E. coli: engineering autotrophic CO2 fixation

大肠杆菌的代谢适应性：工程自养二氧化碳固定

• 批准号: 8311290
• 负责人: Wade M Hicks
• 金额: $ 4.92万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8311290
• 关键词: Assimilations; Atherosclerosis; Bacteria; Biochemical Pathway; Carbon; Carbon Dioxide; Consumption; Coupling; Cyanobacterium; Development; Diabetes Mellitus; Disease; Engineering; Environmental Protection; Enzymes; Escherichia coli; Evolution; Gene Targeting; Genes; Goals; Growth; Health; Human; Knowledge; Light; Malignant Neoplasms; Measurement; Metabolic; Metabolic Pathway; Metabolism; Methods; Modeling; Mutation; Obesity; Organism; Pathway interactions; Production; Property; Regulation; Scheme; Source; atmospheric carbon dioxide; base; carbon fixation; directed evolution; enzyme deficiency; genome sequencing; improved; insight; metabolomics; planetary Atmosphere; reconstitution; remediation; sample fixation

DESCRIPTION (provided by applicant): The wealth of knowledge about central carbon metabolism has allowed organisms to be engineered for the production of useful molecules and has provided insight into many diseases, including obesity, diabetes, atherosclerosis, and some cancers (1, 2). Further advances in metabolic engineering have the potential to significantly impact both environmental protection and human health. For example, CO2 levels in the atmosphere are rising as human consumption of carbon-based fuels increases (3, 4), and the consequences on a 50-year timescale are expected to be dramatic. The goal of this proposal is to improve our understanding of central carbon metabolism by determining the changes needed to convert an organism from a heterotrophic to autotrophic mode of growth. Specifically, I propose to convert the well-characterized heterotrophic bacterium E. coli to an autotrophic mode of growth in which it fixes CO2 as its sole carbon source, using the Calvin cycle. This will involve introduction of foreign genes, targeted mutation of endogenous genes, quantitative modeling and measurement of metabolic fluxes, and directed evolution. The results will provide information about the regulation, evolution and plasticity of central carbon metabolism, and may enable new methods for carbon sequestration from the atmosphere. PUBLIC HEALTH RELEVANCE: Furthering our understanding of how the E. coli metabolic network can adapt to major changes will have at least a two-fold benefit. First, we hope to shed light on how bacterial metabolism can adapt to compensate for major changes to key steps of carbon utilization. Secondly, we hope to use this knowledge to drive the development of a rationally engineered strain of E. coli capable of surviving solely on carbon dioxide for the purpose of coupling atmospheric CO2 remediation with the production of molecules with medicinal or other useful properties.

描述（由申请人提供）：有关中央碳代谢的丰富知识使生物可以用于生产有用分子的生产，并为包括肥胖，糖尿病，动脉粥样硬化和一些癌症等许多疾病提供了见解（1，2）。代谢工程的进一步进步有可能显着影响环境保护和人类健康。例如，随着人类对碳基燃料的消费量的增加，大气中的二氧化碳水平正在上升（3、4），预计50年时间尺度的后果将是戏剧性的。该提案的目的是通过确定将生物从异养和自养生长模式转化所需的变化，以提高我们对中央碳代谢的理解。具体而言，我建议将特征良好的异嗜性细菌大肠杆菌转换为一种自养生长模式，其中使用加尔文循环将其固定为二氧化碳作为其唯一的碳源。这将涉及外国基因的引入，内源基因的靶向突变，代谢通量的定量建模和测量以及定向进化。结果将提供有关中央碳代谢的调节，演变和可塑性的信息，并可能为来自大气中的碳固换提供了新的方法。 公共卫生相关性：增进我们对大肠杆菌代谢网络如何适应重大变化的理解，至少将具有两倍的好处。首先，我们希望阐明细菌代谢如何适应以补偿对碳利用的关键步骤的重大变化。其次，我们希望利用这些知识来开发能够仅在二氧化碳生存的大肠杆菌的合理工程菌株，以便将大气二氧化碳补救与具有药用或其他有用特性的分子产生耦合。