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

在合成化学中，利用工程酶来进行化学转化已经越来越突出。这些生物催化剂很有吸引力，因为它们在产量和选择性方面提供了高效的合成，同时由于对卤化或金属原料的依赖最小而具有固有的环境可持续性。特别是，目前人们对将生物催化反应整合到更大的多步合成方案中以实现分子的“伸缩”结构非常感兴趣。在需要快速构建复杂分子（如制药、农用化学品）的化学工业中，这是一个特别感兴趣的领域，但该领域的研究仍处于起步阶段。例如，我们最近展示了两种聚合生物催化反应的组合：一种是过氧化物酶介导的儿茶酚氧化成醌，另一种是经过卤化酶催化的溴化和/或氯化的芳香胺——所有这些反应都在一个容器中进行。目前特别感兴趣的是原位生成邻醌类化合物（QM），这是化学合成中的多功能中间体。本项目将通过对反应条件的修改和酶的诱变工程，进一步探索这些QMs的生物催化生成，并开发“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这个项目的主要目标。例如，开发具有高氧化还原电位的酶（使非活性芳香族化合物能够氧化），并且能够抵抗极端pH值和高浓度有机溶剂。因此，学生将在有机化学（生物有机化学）和分子生物学（酶学）之间的界面工作。因此，他们将获得广泛的专业知识，这对工业生物技术部门至关重要-通过对生物分子结构和化学反应性的基本见解，合理地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.