Discovery and Development of New Enzymes for Biocatalysis in Organic Synthesis

用于合成的新酶的产生和进化是一个令人兴奋的新研究领域。非常低的初始活性就是进化出新的非天然生物催化剂所需的全部起点。一个合适的高通量筛选方法的可用性对于酶的定向进化是至关重要的。卡内尔团队最近开发了一种醛氧化酶(PaoABC)的比色固相屏幕，使我们能够一次筛选数千个突变。在这个项目中，我们计划开发我们的新型平板筛选，通过突变PaoABC来进化新的酶反应。这种酶是称为黄嘌呤氧化还原酶(XOR)的家族的一部分，该家族对包括醛、嘌呤和亚胺在内的一系列底物使用相同的催化化学。我们的目标是开发这些酶，并将它们集成到合成酶级联中，用于合成生物活性分子、生物质转化和醛的对映选择性氧化。

The creation and evolution of new enzymes for synthesis is an exciting new area of research. A very low initial activity is all that is needed as a starting point to evolve new non-natural biocatalysts. The availability of a suitable high throughput screening method is essential for directed evolution of enzymes. The Carnell group has recently developed a colorimetric solid phase screen for the enzyme aldehyde oxidase (PaoABC) enabling us to screen several thousand mutants at a time. In this project we plan to exploit our novel plate screen to evolve new enzyme reactions by mutation of PaoABC. This enzyme is part of a family called xanthine oxidoreductases (XOR) that use the same catalytic chemistry for a spectrum of substrates including aldehydes, purines and imines. Our intention is to develop these enzymes and integrate them into synthetic enzyme cascades for the synthesis of bioactive molecules, for biomass conversion and for enantioselective oxidation of aldehydes.