Engineering spatial segregation within bacterial hosts for bio-therapeutic protein production

在重组宿主中生产工业感兴趣的生物治疗蛋白正成为一种日益流行的替代方法，可以在哺乳动物细胞培养中生产，这可能是缓慢和昂贵的。该项目的总体目标是通过在大肠杆菌细胞质内进行工程区隔化，开发一种用于重组蛋白生产的细菌系统，这可能会为促进翻译后修饰蛋白的生产和纯化提供环境。在自然界中，区隔化是通过将酶定位在称为细菌微室（BMCs）的复杂蛋白质细胞器内实现的。在过去的十年中，在设计这些bmc方面取得了重大进展，允许在细菌细胞内构建合成微室。在这个项目中，这些结构用于生产和纯化重组蛋白的潜力进行了探索。该项目将扩大我们对工程合成区隔化基础知识的理解，蛋白质区隔化的好处和局限性，并为未来微区隔衍生技术的合理设计提供信息。该项目与EPSRC生物工艺工程领导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，并将影响成本效益和可持续生物基制造的发展。

The production of bio-therapeutic proteins of industrial interest in recombinant hosts is becoming an increasingly popular alternative to production in mammalian cell culture which can be slow and expensive. The overall aim of this project is to develop a bacterial system for recombinant protein production by engineering compartmentalisation within the cytoplasm of E. coli which may allow for an environment that promotes production and purification of post-translationally modified proteins. In nature, compartmentalisation is achieved by localising enzymes inside of sophisticated proteinaceous organelles called bacterial microcompartments (BMCs). In the past decade, significant progress has been made to engineer these BMCs, to permit the construction of synthetic microcompartments within the bacterial cell. In this project the potential of these structures to be used for the production and purification of recombinant proteins is explored.The project will expand our understanding of the fundamentals of engineering synthetic compartmentalisation, the benefits and limitations of protein compartmentalisation 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.