Generalised Photocatalysis by Enzymes (GENPENZ)

酶催化正在以惊人的速度工业化，提供避免昂贵的重金属催化剂，高温和高压的化学转化途径，并提供令人印象深刻的对映体，区域和化学选择性。简而言之，生物催化剂是生物经济的基石：它们在活细胞或无细胞制剂中单独或级联使用，以每天制造化学品，材料，医疗保健产品，燃料和药品;它们是许多诊断和工业传感应用的组成部分。它们是支撑循环经济的技术的核心组成部分，并为实现净零增长、清洁增长和生物经济等全球挑战提供了工程生物学途径。快速和可预测地开发和定制生物催化剂活性的能力对于实现当代全球挑战和英国政府的创新战略至关重要。尽管天然酶和工程酶至关重要，但绝大多数天然酶和工程酶都是热活化的。这种对热活化催化的依赖性：i）将生物催化限制在生物学中天然存在的那些反应类型; ii）高度依赖昂贵且不稳定的辅因子/辅酶;以及iii）对能源的提供提出相当大的需求（生物化学/人工还原剂），“生物反应器”设计（例如在无细胞形式、纳米级装置或微生物细胞工厂内）;和iv）限制调节生物催化剂/生物过程活性的方法。利用光来驱动酶催化将绕过许多这些障碍。然而，除了三个已知的例外，自然界并不使用酶促降解。因此，生物学无法获得广泛的“难以实现”的反应，这些反应将在催化科学中产生变革，以及这些反应在现代世界中的应用。光是免费提供和非侵入性的，但天然辅因子如黄素的光化学多功能性很少被酶使用。因此，确保预测性光生物催化设计的通用路线是一个基本的生物挑战。如果成功的话，确定光生物催化剂工程和设计的通用路线将对新兴生物经济中的催化科学产生变革性影响。该项目将通过使用黄素的天然光化学来解决这一迫切需求，使任何含黄素的蛋白质都有可能被吸收。该计划（称为GENPENZ）位于生物学和酶设计和工程的前沿。它将利用现有（自上而下）和人造（自下而上）蛋白质支架来概括光生物催化剂设计和工程的概念，以生物编码具有广泛反应范围的新光生物催化剂，或从合成肽中重新组装蛋白质框架。它将在12个十年的时间（fs - s）内将可见/红外光谱区域的时间分辨1D / 2D光谱与光质谱（离子迁移率;氢氘交换），EPR光谱和光生物催化剂设计工程的新兴能力结合起来。高级计算化学将支撑所有蛋白质设计/工程工作，光谱学和结构解析。GENPENZ基于与酶促反应机制相关的发现科学的突破。实现光生物催化剂设计的通用平台将开辟新的高能反应途径，丰富催化结果，并避开新兴生物经济中热活化生物催化的许多科学/经济限制。

Enzyme catalysis is being industrialised at a phenomenal rate, offering routes to chemical transformations that avoid expensive heavy metal catalysts, high temperatures and pressures, and providing impressive enantio-, regio- and chemo-selectivities. In short, biocatalysts are a cornerstone of the bioeconomy: they are required individually, or as cascades, in live cells or cell-free preparations to manufacture every day chemicals, materials, healthcare products, fuels and pharmaceuticals; and they are integral to many diagnostic and industrial sensing applications. They are central components of technologies underpinning the circular economy and offer engineering biology routes to realising global challenges, including net zero, clean growth and the bioeconomy. An ability to exploit and tailor biocatalyst activities both rapidly and predictably is essential to realising the contemporary global challenges and the UK Government's Innovation Strategy.Despite their central importance, the vast majority of natural and engineered enzymes are thermally-activated. This dependence on thermally-activated catalysis: i) limits biocatalysis to those reaction types found naturally in biology; ii) places a high dependence on expensive and unstable cofactors / coenzymes; and iii) places a sizeable demand on the provision of energy source (biochemical / artificial reductants), 'bioreactor' designs (e.g. within cell-free formats, nanoscale devices or microbial cell factories); and iv) restricts approaches to regulating biocatalyst / bioprocess activity. The use of light to drive enzyme catalysis would bypass many of these hurdles. However, with only three known exceptions, nature does not make use of enzymatic photocatalysis. Therefore, biology cannot access a broad range of 'difficult-to-achieve' reactions that would be transformational in catalysis science, and applications of these reactions in the modern world. Light is freely available and non-invasive, yet the photochemical versatility of natural cofactors such as flavin is seldom used by enzymes. Therefore, securing generalised routes to predictive photobiocatalysis design is a fundamental biological challenge. If successful, identifying generalised routes to the engineering and design of photobiocatalysts would be transformative for catalysis science in the emerging bioeconomy. This project will address this urgent need by using the natural photochemistry of flavin to make possible photocatalysis by any flavin-containing protein. This programme (termed GENPENZ) is positioned at the frontier of biological photocatalysis and enzyme design and engineering. It will generalise the concept of photo-biocatalyst design and engineering using existing (top down) and man-made (bottom up) protein scaffolds to biologically encode new photo-biocatalysts with wide reaction scope, or to assemble de novo protein frameworks from synthetic peptides. It will unite time-resolved 1D / 2D spectroscopy in the visible / infra-red spectral regions, across 12 decades of time (fs - s), with emerging capabilities in photo mass spectrometry (ion mobility; hydrogen-deuterium exchange), EPR spectroscopy, and photo-biocatalyst design engineering. High-level computational chemistry will underpin all protein-design/engineering work, spectroscopy, and structure elucidation. GENPENZ is based on breakthroughs in discovery science relating to mechanisms of enzyme photocatalysis. Realisation of a generalised platform for photo-biocatalyst design will open up new high-energy reaction pathways, enrich catalysis outcomes, and sidestep many of the scientific / economic constraints of working with thermally-activated biocatalysis in the emerging bioeconomy.