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

用于微滴酶进化实验的超灵敏、超快吸收光谱仪

基本信息

批准号：
BB/R022178/1
负责人：
Frank Vollmer
金额：
$ 19.21万
依托单位：
UNIVERSITY OF EXETER
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2018
资助国家：
英国
起止时间：
2018 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FR022178%2F1
关键词：
Ultra Sensitive Fast Absorption Spectrometer

项目摘要

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.

大多数生物过程是由酶的功能驱动的，这些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。我们的细胞是由各种各样的酶功能驱动的，这些功能决定了从细胞如何移动和生长到细胞如何将能量转化为可用形式的一切。许多生物、医学和工业科学都专注于了解酶的工作原理，并发现新的酶，为我们的医学和工业应用提供新的功能。然而，我们研究酶功能的能力往往局限于集中研究，包括基因/蛋白质的一些微小变化，而且，集中在一个相对较小的池以前表征的酶。我们现在需要的是新的技术，使我们能够研究和比较酶的功能，并使我们能够对这种酶进行大量的改变。此外，这些技术还应该使我们能够筛选大量的蛋白质，以便在各种各样的未知基因中寻找新型酶。这只能通过开发新技术来实现，使我们能够同时对数百万个基因进行采样和研究，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形成的，DNA中记录的遗传密码的性质最终决定了酶的性质及其工作方式。我们将开发一种新技术，使我们能够同时将数千个DNA片段表达为蛋白质。然后，我们将使用尖端的微管道将每个具有不同类型DNA/蛋白质的单细胞分离到一个单独的隔室中作为液滴。使用最先进的方法来研究化学反应，我们将建立一种新的方法来检测每个微室中的化学变化。这种具有创新性和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。所提出的光流体技术的力量将使我们能够进行超越当前方法的高灵敏度测量。具体来说，用于检测化学变化的新的光流体技术将比目前使用的技术灵敏1000倍左右。通过将其与基于液滴的划分相结合，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，这些方法既可以研究酶进化与酶功能的关系，也可以发现新的酶，还可以开发新的抗微生物药物。