Towards vibrational control of enzymes for biotechnology and biocatalysis

绝大多数生物反应都是由酶催化的，酶通常表现出极高的选择性和催化速率的大幅提高。随着酶工程的进步和对更可持续的合成的日益增长的需求，酶介导的生物催化正在成为商品和高价值化学品生产的传统方法的既定替代方案。在许多情况下，期望具有快速且实时的调节酶活性的方法，例如以控制通过多酶级联或体内代谢途径的通量。一种可能性是通过酶反应的直接振动控制。化学反应通常涉及键的形成和断裂，这伴随着反应物和周围环境的分子振动的变化，因此可以通过激发促进或降级振动模式来改变反应速率。在凝聚态物质（蛋白质、表面等）中，来自激发分子振动的能量预计会在皮秒内消散，由于导致样品加热，连续激发是不可能的。超快激光光谱学的最新进展提供了一种解决方案-现在可以使用红外（IR）光的超快（亚皮秒）脉冲直接激发分子振动，这可以导致气相和表面上的热反应的显着速率增强，而不会过度加热。该项目现在的目标是扩展和发展这种方法，使用超快IR光脉冲来控制酶反应的速率。这项工作将涉及仪器和方法开发以及新的数据分析和实验设计程序。计算化学（MD模拟和/或DFT建模）将用于辅助实验设计和结果解释。最终，我们将设计和测试一种新的超快激光控制生物反应器。该项目福尔斯属于“技术和方法开发”和“工业生物技术”的职权范围，并牢固地嵌入化学，生物学和物理学的界面，这是BBSRC在“探索新的工作方式”议程中的关键驱动力。它借鉴了关键的生物科学技能，包括适用于生物科学的新领域（例如蛋白质工程，动态结构科学）和最先进的时间分辨光谱方法。这些都是“风险”的技能在英国的景观和培训，因此，在这些领域是至关重要的，以确保平衡和知识渊博的人才库是在英国生物科学保持。总的来说，这项工作将提供一个高度跨学科的方法（生物）化学/生物制药学为基础的研究，提供高度多样化的培训机会，博士生，谁将有额外的好处，能够每天访问主管的实验室，因为他们都位于同一栋楼（MIB）。

The vast majority of biological reactions are catalysed by enzymes, which often exhibit exquisite selectivity and massive catalytic rate enhancements. With advances in enzyme engineering and an increasing desire for more sustainable synthesis, enzyme-mediated biocatalysis is becoming an established alternative to traditional methods of both commodity and high-value chemical production. In many cases, it would be desirable to have a rapid and real-time method of tuning enzyme activity, e.g. to control flux through a multi-enzyme cascade or in vivo metabolic pathway. One possibility is through direct vibrational control of the enzyme reaction. Chemical reactions typically involve the making and breaking of bonds, which are accompanied by changes in molecular vibrations of the reactants and surrounding environment, so it is possible to alter the rate of reaction through excitation of promoting or demoting vibrational modes. In condensed matter (proteins, surfaces, etc) energy from excited molecular vibrations is expected to dissipate in picoseconds, making continuous excitation impossible due to resulting sample heating. Recent advances in ultrafast laser spectroscopy allows a solution - It is now becoming practical to directly excite molecular vibrations using ultrafast (sub-picosecond) pulses of infrared (IR) light, which can lead to significant rate enhancements of thermal reactions in the gas-phase and on surfaces without excessive heating. This project now aims to extend and develop this methodology, using ultrafast IR light pulses to control the rate of enzyme reactions.This work will involve instrument and method development alongside new data analysis and experimental design procedures. Computational chemistry (MD simulations and/or DFT modelling) will be used to aid in experimental design and interpretation of results. Ultimately, we will aim to design and test a new ultrafast laser-controlled bioreactor. The project falls within the remit of both 'technologies and methodological development' and 'industrial biotechnology' and is firmly embedded at the interface of chemistry, biology and physics, a key driver for BBSRC in the 'Exploiting new ways of working' agenda. It draws on key bioscience skills, including new areas of applicable to bioscience (e.g. protein engineering, dynamic structural science) and state-of-the-art time resolved spectroscopy approaches. These are 'at risk' skills in the UK landscape and training in these areas is therefore vital to ensure a balanced and knowledgeable talent pool is maintained in the UK biosciences. Overall, the work will provide a highly interdisciplinary approach to (bio)chemistry/ biophysics-based research, offering highly diverse training opportunities to a PhD student, who will have the additional benefit of being able to access the supervisors' laboratories on a daily basis, as they are all co-located within the same building (MIB).