Development and Application of Next Generation Synthetic Biology Tools

对包括E. coli和革兰氏阳性菌S.酿酒酵母（面包酵母）用于商业生物生产过程以制造多种产品，范围从高价值低容量产品，例如生物治疗剂（例如重组胰岛素）到中等价值产品（例如生物催化剂和专用化学品），到低价值的大宗商品产品（例如琥珀酸和生物燃料）。此外，这些生产过程中使用的生物技术也常用于基础生命科学研究。因此，开发新的基因控制系统将在生物学和生物医学研发的许多应用和基础领域中具有极大的益处。特别是，本研究旨在开发新的蛋白质生产和代谢工程工具，并展示这些新的合成生物学工具在生物加工工业中的应用。在制药行业，生物制药在10种最畅销产品中占了7种。2010年，全球蛋白质生物制药市场的销售额为700亿美元，到2015年，这一数字将增至1100亿美元。因此，预计全球销售量增加的原因是：i）这些治疗性蛋白质的许多靶标被认为是传统小分子方法“无法控制的”，ii）治疗性蛋白质通常具有上级的安全性和功效特征，iii）治疗性蛋白质的平均开发时间比传统疗法短，iv）通常存在有限或缺乏竞争。然而，尽管生物制药提供了许多健康益处沿着大量的商业机会，但其生产仍然是一个重大的技术挑战。通过目前的研究，我将开发并展示新型基因共表达技术的四个重要特征，以使多聚体蛋白质产品能够更有效地生产，沿着提供更简单、更有效的生产工艺。此外，这些共表达技术将用于优化多个多元共表达挑战，有助于指导代谢工程的努力。这项研究的成果将提高生物工艺的效率，有可能减少药物开发时间和生产成本，从而减轻国家医疗保健提供者的财政负担。该研究项目还将产生有助于保持英国竞争优势的技术和知识，并将培养训练有素的熟练研究人员。

A variety of micro-organisms including E. coli and S. cerevisiae (baker's yeast), are used in commercial bio-production processes to manufacture a number of products, ranging from high-value low-volume products, such bio-therapeutics (e.g. recombinant insulin), to mid-value products, (e.g. biocatalysts and specialised chemicals), to low-value bulk commodity products (e.g. succinic acid and biofuels). In addition, the same biotechnologies used in these production processes are commonly used in basic life science research. As such, the development of new gene control systems would be of great benefit in a number of applied and fundamental areas of biological and biomedical R&D. In particular, the research here seeks to develop novel protein production and metabolic engineering tools, and to demonstrate the applications of these novel synthetic biology tools in the context of the bioprocessing industry. Within the pharmaceutical sector, biopharmaceuticals constitute 7 out of the 10 bestselling products. The global protein-based biopharmaceutical market had sales worth $70b in 2010, and this figure is set to increase to $110b by 2015. The reasons for the increase in projected global sales are thus: i) many of the targets of these therapeutic proteins are deemed 'undruggable' by traditional small molecule approaches, ii) therapeutic proteins often have superior safety and efficacy profiles, iii) the development times on average are shorter for therapeutic proteins than traditional therapies, iv) there is often limited or absent competition. However, although biopharmaceuticals offer many health benefits along with substantial commercial opportunities, their production remains a significant technical challenge. Through this current study, I shall develop and demonstrate four important flavours of a novel gene co-expression technology, to allow multimeric protein products to be produced more effectively, along with the potential to provide a simpler and more efficient manufacturing process. Additionally, these co-expression technologies will be used to optimise a number of multivariate co-expression challenges, helping to guide metabolic engineering efforts. The outputs from this fellowship will lead to improved bioprocess efficiencies, with the potential to reduce both drug development times and manufacturing costs, and therefore the financial burden upon national healthcare providers. This research project will also generate technology and knowledge that will help maintain the UK's competitive edge, and will produce highly trained and skilled research personnel.