Towards vibrational control of enzymes for biotechnology and biocatalysis

绝大多数生物反应都是由酶催化的，酶往往表现出极高的选择性和巨大的催化效率。随着酶工程的进步和人们对更可持续的合成的日益渴望，酶介导的生物催化正在成为传统的商品和高价值化学生产方法的替代方法。在许多情况下，希望有一种快速和实时的方法来调节酶的活性，例如通过多酶级联或体内代谢途径来控制通量。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建立和断裂，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低振动模式来改变反应速度。在凝聚态物质(蛋白质、表面等)中，激发的分子振动能量预计将在皮秒内消散，从而由于产生的样品加热而不可能连续激发。超快激光光谱学的最新进展提供了一种解决方案-使用超快(亚皮秒)红外(IR)光脉冲直接激发分子振动现在变得可行，这可以在不过度加热的情况下显著提高气相和表面上的热反应速度。这个项目现在的目标是扩展和发展这一方法，使用超快红外光脉冲来控制酶反应的速度。这项工作将涉及仪器和方法的开发，以及新的数据分析和实验设计程序。将使用计算化学(MD模拟和/或DFT建模)来协助实验设计和结果解释。最终，我们的目标是设计和测试一种新的超快激光控制的生物反应器。该项目属于“技术和方法开发”和“工业生物技术”的范畴，并牢固地植根于化学、生物和物理的交界处，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学方法。在英国，这些技能都是“危险的”技能，因此，这些领域的培训对于确保英国生物科学领域维持一个平衡和知识渊博的人才库至关重要。总体而言，这项工作将为基于(生物)化学/生物物理学的研究提供高度跨学科的方法，为博士生提供高度多样化的培训机会，他们将获得额外的好处，能够每天进入主管的实验室，因为他们都位于同一大楼(MIB)内。

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).