Building synthetic enzyme fusions for chiral molecule production.

构建用于手性分子生产的合成酶融合体。

• 批准号: 1618783
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1618783
• 关键词: Building; synthetic; enzyme; fusions; chiral

Strategic Research Priority: World Class Underpinning ResearchOptically active amines play an important role as bioactive compounds in pharmaceutical and chemical industries. Because classical chemical approaches are complex and expensive, biocatalytic routes are desirable. One route for their bio-production involves conversion of alcohols to amines. Currently, there are no known enzymes that can convert alcohols directly to amines, however a two-step transformation is possible via alcohol oxidation and a subsequent reductive amination or transamination.In enzyme cascades, reaction engineering is necessary to ensure flux towards the products, and to avoid accumulation of, often, reactive intermediates. In contrast, naturally occurring multifunctional enzymes have evolved to keep reactive substrates within enzyme complexes until key transformations have been carried out. In vivo, multifunctional enzymes are thought to confer selective advantage via increased reaction rates and chemical stability or prevention of toxicity from reactive intermediates.The aim of this project was to characterise a natural fusion between a transaminase and a dehydrogenase, protein PP_2782, in Pseudomonas putida KT2440 (a non-pathogenic soil bacterium that has a diverse range of amino and fatty acid metabolic pathways), in order to exploit its biosynthetic abilities for novel enzyme fusion engineering. Using this natural fusion as a guide template, and to enable the synthesis of novel bioactive compounds that are challenging to produce chemically, a number of synthetic fusions comprising well-characterised enzymes was engineered and tested.

战略研究重点：光学活性胺作为生物活性化合物在医药和化工领域发挥着重要作用。由于经典的化学方法复杂且昂贵，因此生物催化途径是理想的。其生物生产的一种途径涉及将醇转化为胺。目前，还没有已知的酶可以将醇直接转化为胺，但是通过醇的氧化和随后的还原胺化或转胺化可以进行两步转化。在酶级联中，反应工程是必要的，以确保流向产物的通量，并避免反应中间体的积累。相比之下，天然存在的多功能酶已经进化为将反应性底物保持在酶复合物内，直到进行关键转化。在体内，多功能酶被认为通过增加反应速率和化学稳定性或防止反应中间体的毒性而赋予选择性优势，本项目的目的是在恶臭假单胞菌KT 2440中表达转氨酶和脱氢酶之间的天然融合蛋白PP_2782（一种具有多种氨基和脂肪酸代谢途径的非致病性土壤细菌），以利用其生物合成能力进行新型酶融合工程。使用这种天然融合物作为指导模板，并且为了能够合成具有化学生产挑战性的新型生物活性化合物，工程化并测试了许多包含充分表征的酶的合成融合物。