Biochemical characterisation of the Glycyl Radical Enzyme Containing Microcompartment (GRM2) from Proteus mirabilis

奇异变形杆菌中含有甘氨酰基酶的微区室 (GRM2) 的生化特性

• 批准号: 1930945
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1930945
• 关键词: Biochemical; characterisation; Glycyl; Radical; Enzyme

The production of bio-therapeutic proteins, small molecule drugs and chemical intermediates of industrial interest in recombinant hosts is becoming an increasingly popular alternative to production in mammalian cell culture, natural hosts (e.g. plants, fungi) or chemical synthesis, which can be slow and expensive. Achieving high product yields in recombinant synthetic pathways requires that the fluxes of each enzymatic reaction of the pathway of interest be balanced to limit the accumulation of intermediates, particularly those toxic to the host. Conventional strategies include the modulation of expression levels of individual enzymes or directed evolution of rate-limiting enzymes and have focused on balancing pathway flux, but don't consider other crucial factors such as diffusion, transport, localisation of intermediates and the cost-effective purification and recovery of products. Nature has developed strategies to deal with many of these issues by confining certain biochemical pathways to various organelles. In prokaryotes, bacterial microcompartments (BMCs) encapsulate enzymes associated with certain metabolic processes within a large protein shell, bringing sequentially acting enzymes into spatial proximity thus increasing local enzyme concentration, protecting the cell from potentially reactive intermediates and facilitating co-factor recycling. In the past decade significant progress has been made to engineer BMCs to permit the construction of BMC bioreactors with novel functions within the cell. The overall aim of this project is to develop BMCs as synthetic biology platforms for spatial organization of enzymes for enhanced production of valuable compounds in E. coli. The project will expand our understanding of the fundamentals of engineering synthetic compartmentalisation, the benefits and limitations of pathway localisation and inform the rational design of future microcompartment-derived technologies. The project is aligned with the EPSRC centre for Doctoral Training in Bioprocess Engineering Leadership. It provides new synthetic biology approaches to the redesign of cellular chemistry and will impact in the development of cost-effective and sustainable bio-based manufacture.BMC tools and technologies will be developed with industrial applications in mind. The work is supported by in-kind contribution from the industrial biotechnology company Ingenza, a world leader in the application of synthetic biology for the provision of efficient bioprocesses for the manufacture of chemicals, biologics, pharmaceuticals and biofuels. Compartmentalisation of enzymes/pathways may provide a valuable strategy for the improvement of industrial strains of interest. The industrial partner will offer enzymes/pathways of interest and advice/consultancy on how industry would like to see the BMC technology applied.

在重组宿主中生产生物治疗蛋白质、小分子药物和具有工业意义的化学中间体正成为一种日益流行的替代方案，替代哺乳动物细胞培养、天然宿主（例如植物、真菌）或化学合成中的生产，这些方法可能缓慢且耗时。昂贵的。在重组合成途径中实现高产物产量需要平衡感兴趣途径的每个酶促反应的通量，以限制中间体的积累，特别是那些对宿主有毒的中间体。传统的策略包括调节单个酶的表达水平或限速酶的定向进化，并侧重于平衡途径通量，但没有考虑其他关键因素，例如扩散、运输、中间体的定位和经济有效的纯化和产品的回收。大自然已经制定了通过限制各种细胞器的某些生化途径来解决许多这些问题的策略。在原核生物中，细菌微区室（BMC）将与某些代谢过程相关的酶封装在一个大的蛋白质壳内，使顺序作用的酶在空间上接近，从而增加局部酶的浓度，保护细胞免受潜在反应中间体的影响并促进辅因子回收。在过去的十年中，BMC 工程取得了重大进展，能够构建在细胞内具有新功能的 BMC 生物反应器。该项目的总体目标是开发 BMC 作为合成生物学平台，用于酶的空间组织，以提高大肠杆菌中有价值化合物的产量。该项目将扩大我们对工程合成区室化基础知识、通路定位的优点和局限性的理解，并为未来微区室衍生技术的合理设计提供信息。该项目与 EPSRC 生物过程工程领导力博士培训中心保持一致。它为细胞化学的重新设计提供了新的合成生物学方法，并将影响具有成本效益和可持续的生物基制造的发展。BMC 工具和技术将在开发时考虑到工业应用。这项工作得到了工业生物技术公司 Ingenza 的实物捐助的支持，该公司是合成生物学应用领域的世界领先者，为化学品、生物制品、药品和生物燃料的制造提供高效的生物工艺。酶/途径的区室化可能为改进感兴趣的工业菌株提供有价值的策略。工业合作伙伴将提供感兴趣的酶/途径以及工业界希望如何应用 BMC 技术的建议/咨询。