Design and Evolution of Photoenzymes for Triplet Energy Transfer Catalysis

三重态能量转移催化光酶的设计和进化

• 批准号: EP/Y023722/1
• 负责人: Anthony Green
• 金额: $ 215.83万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FY023722%2F1
• 关键词: Design; Evolution; Photoenzymes; Triplet; Energy

The ability to program new modes of catalysis into enzymes would have profound impacts across chemistry and biotechnology, delivering sustainable biocatalytic processes to address societal needs. Genetic code expansion technology opens up exciting new opportunities in biocatalyst design and engineering, by allowing site-selective introduction of new functional elements into proteins as non-canonical amino acid side chains. In TRIPase, I will exploit an expanded genetic code to develop efficient, selective and programmable photoenzymes that operate via triplet energy transfer processes, a versatile mode of reactivity in organic synthesis that is currently not accessible to biocatalysis. These photoenzymes will contain functional elements needed to harness light energy and promote valuable chemical reactions including [2+2]cycloadditions, electrocyclizations, rearrangements and deracemizations. My approach will employ engineered translation components to introduce organic photosensitizers into protein active sites. Protein cavities offer attractive and versatile chiral environments for mediating enantioselective photochemistry, where substrates and key catalytic elements can be accurately positioned within a single pocket. Since the photosensitizers are genetically encoded, active photoenzymes can be optimized via directed evolution to enhance catalytic efficiency and quantum yields, or to impart new functions that are challenging to achieve with small molecule photocatalysts. Structural and biochemical analysis of engineered photoenzymes will shed light on the active site features and mechanistic strategies responsible for enhanced photocatalysis to guide future biocatalyst design. Overall, the platform technology developed in TRIPase will open the door to a wealth of new excited-state chemistry in proteins and in doing so will underpin the development of a new generation of evolvable photocatalysts with efficiencies and specificities akin to natural enzymes.

将新催化模式编程为酶的能力将在化学和生物技术方面产生深远的影响，从而提供可持续的生物催化过程，以满足社会需求。遗传代码扩展技术通过允许将新的功能元素引入蛋白质作为非经典氨基酸侧链，从而为生物催化剂设计和工程开辟了令人兴奋的新机会。在 TRIPase 中，我将利用扩展的遗传密码来开发高效、选择性和可编程的光酶，这些光酶通过三重态能量转移过程进行操作，这是有机合成中的一种多功能反应模式，目前生物催化无法实现。这些光酶将含有利用光能和促进有价值的化学反应（包括[2+2]环加成、电环化、重排和去消旋化）所需的功能元素。我的方法将采用工程翻译组件将有机光敏剂引入蛋白质活性位点。蛋白质空腔为介导对映选择性光化学提供了有吸引力且多功能的手性环境，其中底物和关键催化元素可以准确地定位在单个口袋内。由于光敏剂是基因编码的，因此可以通过定向进化来优化活性光酶，以提高催化效率和量子产率，或赋予小分子光催化剂难以实现的新功能。工程光酶的结构和生化分析将揭示负责增强光催化作用的活性位点特征和机制策略，以指导未来的生物催化剂设计。总体而言，TRIPase 开发的平台技术将为蛋白质中大量新的激发态化学打开大门，从而支持新一代可进化光催化剂的开发，其效率和特异性类似于天然酶。