Engineered oxidative enzymes as catalysts for the formation of quinone methides in integrated chemical synthesis.

在合成化学中，使用工程化酶来执行化学转化已经越来越突出。这些生物催化剂是有吸引力的，因为它们在产率和选择性方面提供了高效的合成，以及源自对卤化或金属原料的最小依赖的固有环境可持续性。特别是，目前有很大的兴趣，在整合生物催化反应到更大的多步合成方案，使“套叠”的分子建设。这是一个在化学工业中特别感兴趣的领域，其中需要快速构建复杂分子（例如药物，农用化学品），但该领域的研究仍处于起步阶段。例如，我们最近已经证明了两个收敛的生物催化反应的组合：过氧化物酶介导的儿茶酚氧化为醌，这是耦合与芳香胺，已经历卤化酶催化的溴化和/或氯化-所有在一个单一的vessel.当前特别感兴趣的是在原位生成的邻醌甲基化物（QM），这是在化学合成中的通用中间体。该项目旨在进一步探索这些QMs的生物催化生成，并通过修改反应条件以及酶的诱变工程来开发“一锅”反应序列。事实上，开发特别适合合成化学的工程过氧化物酶将是该项目的主要目标。例如，开发具有高氧化还原电位的酶（使不反应的芳香族化合物氧化），以及耐极端pH和高浓度有机溶剂的酶。因此，学生将在有机化学（生物有机化学）和分子生物学（酶学）之间的界面上工作。因此，他们将获得对工业生物技术领域至关重要的广泛专业知识-通过对生物分子结构和化学反应性的基本见解，合理设计新的高价值生物催化剂的能力。

In synthetic chemistry, there has been increasing prominence in the use of engineered enzymes to execute chemical transformations. These biocatalysts are attractive since they offer highly efficient synthesis in terms of yields and selectivity, together with an inherent environmental sustainability stemming from a minimal reliance on halogenated or metallic feedstocks. In particular, there is currently great interest in the integration of biocatalytic reactions into larger multi-step synthetic schemes to enable the "telescoped" construction of molecules. This is an area that is of particular interest in chemical industries where there is a need for the rapid construction of complex molecules (e.g. pharmaceuticals, agrochemicals), yet research in this area is still in its infancy. For example, we have recently demonstrated the combination of two convergent biocatalytic reactions: a peroxidase-mediated oxidation of catechols to quinones, which is coupled with an aromatic amine that has undergone halogenase-catalysed bromination and/or chlorination - all in a single vessel.Of current particular interest is the in situ generation of o-quinone methides (QM), which are versatile intermediates in chemical synthesis. This project will aim to further explore the biocatalytic generation of these QMs and develop the "one-pot" reaction sequences, through modifications of the reaction conditions as well as the mutagenic engineering of the enzymes. Indeed, the development of engineered peroxidase enzymes that are particularly suited for synthetic chemistry will be a major goal of this project. For example, the development of enzymes that have a high redox potential (to enable the oxidation of unreactive aromatic compounds), and that are resistant to extremes of pH and high concentrations of organic solvents.The student will therefore work at the interface between organic chemistry (bioorganic chemistry) and molecular biology (enzymology). They will thus gain a wide range of expertise that are critical for the industrial biotechnology sector - the ability to rationally engineer new, high-value biocatalysts through fundamental insights in biomolecular structure and chemical reactivity.