Design and Evolution of Enzymes with Non-Canonical Catalytic Mechanisms

具有非典型催化机制的酶的设计和进化

• 批准号: 2449667
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2449667
• 关键词: Design; Evolution; Enzymes; Non; Canonical

The ability to rationally design an enzyme for any desired transformation would have major impacts across the pharmaceutical and wider chemical sectors, leading to more efficient and sustainable synthetic routes to high value chemicals. The combination of computational enzyme design and directed evolution is currently the most attractive strategy to achieve this ambition. However, the limited range of functional groups presented by the genetic code (i.e. 20 amino acids) restricts the range of catalytic mechanisms which can be installed into designed active sites, thus severely limiting the repertoire of chemical transformations accessible. Advanced protein engineering technologies available in our laboratory now allow us to install non-canonical 'chemically inspired' amino acids into enzyme active sites, thus greatly expanding upon the limited range of functionality accessible with Nature's genetically encoded residues. We have recently combined this genetic code expansion technology with computational enzyme design and laboratory evolution to create the first enzyme that operates via a non-canonical organocatalytic mechanism (Nature 2019, 570, 219). In this PhD studentship, we will now expand ambitiously upon this early success, to demonstrate that the integration of non-canonical amino acids into computational design and directed evolution workflows can lead to the creation of enzymes with novel catalytic mechanisms and new activities. The design of new functional amino acids will be inspired by small molecule organocatalysts which are able to promote a plethora of valuable transformations not observed in Nature. Our approach merges the complementary disciplines of organocatalysis and biocatalysis, combining the mechanistic and functional versatility of small molecule systems with the enormous rate accelerations and reaction selectivities achievable within evolvable protein scaffolds.The project is firmly embedded at the interface of chemistry, biology and biophysics, a key driver for the BBSRC in the 'Exploiting new ways of working' agenda. It draws on key bioscience skills, including protein engineering, directed evolution and enzyme characterization, and specifically exploits a multi-disciplinary approach to the creation of enzymes with novel function, which is a major objective in the Industrial Biotechnology sector.

为任何期望的转化合理设计酶的能力将对整个制药和更广泛的化学部门产生重大影响，从而产生更有效和可持续的高价值化学品合成途径。计算酶设计和定向进化的结合是目前实现这一目标最具吸引力的策略。然而，遗传密码所提供的有限的官能团（即20个氨基酸）限制了可以安装到设计活性位点的催化机制的范围，从而严重限制了化学转化的可获得性。在我们的实验室中，先进的蛋白质工程技术现在允许我们将非规范的“化学启发”氨基酸安装到酶的活性位点，从而大大扩展了自然遗传编码残基的有限功能范围。我们最近将这种遗传密码扩展技术与计算酶设计和实验室进化相结合，创造了第一种通过非规范有机催化机制起作用的酶（Nature 2019, 570, 219）。在这个博士研究生项目中，我们将在这一早期成功的基础上雄心勃勃的扩展，以证明将非规范氨基酸整合到计算设计和定向进化工作流程中可以导致具有新型催化机制和新活性的酶的创造。新功能氨基酸的设计将受到小分子有机催化剂的启发，这些催化剂能够促进自然界中未观察到的大量有价值的转化。我们的方法融合了有机催化和生物催化的互补学科，将小分子系统的机制和功能通用性与可进化蛋白质支架内可实现的巨大速率加速和反应选择性相结合。该项目牢牢地嵌入了化学、生物学和生物物理学的界面，这是BBSRC“探索新的工作方式”议程的关键驱动力。它利用关键的生物科学技能，包括蛋白质工程、定向进化和酶表征，并特别利用多学科方法来创造具有新功能的酶，这是工业生物技术部门的主要目标。

项目成果

Engineering enzyme activity using an expanded amino acid alphabet.

• DOI: 10.1093/protein/gzac013
• 发表时间: 2023-01-21
• 期刊: PROTEIN ENGINEERING DESIGN & SELECTION
• 影响因子: 2.4
• 作者: Birch-Price, Zachary;Taylor, Christopher J.;Ortmayer, Mary;Green, Anthony P.
• 通讯作者: Green, Anthony P.