Ultra-Sensitive and Ultra-Fast Absorption Spectrometer for Micro-Droplet-based Enzyme Evolution Experiments

大多数生物过程是由酶的功能驱动的，酶是与化合物结合的蛋白质，通过这种相互作用将化合物改变为不同的化学物质。我们的细胞由各种各样的酶功能驱动，这些功能决定了一切，从我们的细胞如何运动和生长，到细胞如何将能量转化为可用的形式。许多生物、医学和工业科学都专注于了解酶是如何工作的，并发现新的酶可以为我们提供医疗和工业应用的新功能。然而，我们研究酶功能的能力往往局限于围绕基因/蛋白质的几个微小变化的重点研究，而且还集中在相对较小的先前表征的酶的池中。我们现在需要的是新的技术，让我们能够研究和比较酶的功能，并允许我们对酶进行大量的改变。此外，这样的技术还应该允许我们筛选大量的蛋白质，以便为新类型的酶搜索广泛多样的未知基因。这只能通过开发新技术来实现，这些技术允许我们同时采样和调查数百万个基因，然后在短时间内进行数百万个酶功能实验。这个项目的目的是开发新技术，使我们能够做到这一点。酶是由DNA上的基因编码的蛋白质组成的，DNA中记录的遗传密码的性质最终决定了酶的性质和它的工作方式。我们将开发一种新技术，使我们能够一次将数千个DNA片段表达成蛋白质。然后，我们将使用尖端的微型管道将每个带有不同类型DNA/蛋白质的单个细胞分离到一个单独的隔间作为液滴。使用最先进的研究化学反应的方法，我们将建立一种新的方法来检测每个微格内的化学变化。这种创新和具有挑战性的技术组合将使我们能够筛选大量的变异酶，并在未知功能的DNA中寻找新的基因功能。提出的光流控技术的力量将使我们能够进行高灵敏度的测量，超过目前的方法。具体地说，将用于探测化学变化的新的光流控技术将比目前使用的技术灵敏约1000倍。通过将其与基于液滴的分区相结合，我们将能够每秒筛选1000个独立的实验。我们提出的项目将打开一系列新方法的大门，既可以研究酶的进化与酶功能的关系，也可以发现新的酶，以及下游新的抗菌药物的开发。

Most biological processes are driven by the function of enzymes, these are proteins that bind to a chemical compound and through this interaction alter the chemical compound into a different chemical substance. Our cells are driven by a huge diversity of enzyme functions that dictate everything from how our cells move and grow to how cells convert energy into a usable form. Much of biological, medical and industrial science is focused on understanding how enzymes work and discovering new enzymes that can provide us with new functions for medical and industrial applications. However, our ability to study enzyme function is often limited to focused studies encompassing a handful of minor changes in the gene/protein and, furthermore, is focused on a relatively small pool of previously characterised enzymes. What we need now is new technologies that allow us to study and compare the function of enzymes and which allows us to make huge numbers of changes to that enzyme. Furthermore, such technologies should also allow us to screen huge numbers of protein in order to search a wide diversity of unknown genes for new types of enzyme. This can only be achieved by developing new technologies that allow us to simultaneously sample and investigate millions of genes and then conduct millions of enzyme function experiments in short time frames. The aim of this project is to develop new technologies that allow us to do this. Enzymes are formed of proteins which are encoded by genes on DNA, the nature of the genetic code recorded in DNA ultimately governs the nature of an enzyme and how it works. We will develop a new technology that will allow us to express thousands of fragments of DNA into protein all at once. We will then use cutting edge micro-plumbing to separate single cells each with different types of DNA/protein into a separate compartment as droplets. Using state of the art methods for studying chemical reactions we will build a new way of detecting chemical changes within each micro-compartment. This innovative and challenging combination of technology will allow us to screen huge numbers of variant enzymes and search DNA of unknown function for new gene functions. The power of the optofluidic technology proposed will allow us to take highly sensitive measurements surpassing current approaches. Specifically, the new optofluidic technology that will be utilised for detecting chemical changes will be around 1000 times more sensitive than current technologies used. By combining it with the droplet based compartmentation we will be able to screen 1000 independent experiments a second. Our proposed project will unlock the door to a range of new approaches for both investigating how enzyme evolution relates to enzyme function, discovery of new enzymes and down-stream the development of new antimicrobial drugs.