Directed Evolution of New Enzymes for Sustainable Chemical Manufacture in Microbial Cells

用于合成类药物分子的新酶的产生使化学制造发生了革命性的变化，特别是在工业规模上。[1]然而，尽管近年来在这一领域取得了重大进展，但并不存在用于每一次有机转化的酶替代。虽然新发现的微生物的测序和环境元基因组的分析将继续填充生物催化工具箱，但这种方法似乎不太可能成为为任何给定的有机反应创建新的生物催化剂的通用解决方案。因此，设计和重新设计现有的酶来执行新的化学转化是有机化学中的一个重要挑战。[2]这个项目将使用定向进化和有机化学的机械知识来创造一个新的酶家族，能够催化(据我们所知)在自然界中从未存在的反应。然后，这些酶将被用作设计代谢途径的模块组件，通过微生物发酵直接从可再生资源中生产制药行业感兴趣的小分子。[3]本项目的高度多学科性质将使学生能够熟练掌握各种化学和生物技术(例如重组DNA技术、聚合酶链式反应、定向进化、微生物学、体外试验开发、基因组工程、多步骤化学合成和光谱分析)。

The creation of new enzymes for the synthesis of drug-like molecules has revolutionised chemical manufacture, especially on the industrial scale.[1] However, despite significant advances in this field in recent years, enzymatic replacements for every organic transformation do not exist. Whilst the sequencing of newly-discovered microorganisms and the analysis of environmental metagenomes will continue to populate the biocatalytic toolbox, it seems unlikely that this approach will ever become a general solution to creating a new biocatalyst for any given organic reaction. The design and re-engineering of existing enzymes to perform novel chemical transformations is therefore an important challenge in organic chemistry.[2]This project will use directed evolution and a mechanistic knowledge of organic chemistry to create a new family of enzymes capable of catalysing reactions that have (to the best of our knowledge) never existed in Nature. These enzymes will then be used as modular components in designer metabolic pathways to produce small molecules of interest to the pharmaceutical industry directly from renewable resources via microbial fermentation.[3]The highly multidisciplinary nature of this project will allow the student to develop a strong proficiency in a variety of chemical and biological techniques (e.g. recombinant DNA techniques, PCR, directed evolution, microbiology, in vitro assay development, genome engineering, multistep chemical synthesis and spectroscopic analysis).