Engineering Enzymes for New Stereoselective and Stereodynamic Processes: An Integrated Chemistry -Bioengineering- X-Ray Crystallography-Molecular Dynamics Approach

用于新立体选择性和立体动力学过程的工程酶：化学-生物工程-X射线晶体学-分子动力学综合方法

• 批准号: 2023250
• 负责人: Mark Wilson
• 金额: $ 60.39万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2023250&HistoricalAwards=false
• 关键词: Engineering; Enzymes; New; Stereoselective; Stereodynamic; Engineering; Enzymes; New; Stereoselective; Stereodynamic

Most pharmaceutical drugs and agrochemicals are what is referred to as fine chemicals. The molecules have very specific structures that give them their beneficial properties. Enzymes are highly effective in producing such structures. The objective of this project is to develop a strategy to design and tune the activity of enzymes that are involved in the production of a class of fine chemicals called taxanes. These compounds have anti-cancer properties. One of the taxanes, taxol, is currently used in the treatment of metastatic breast cancers. The project will include educating and mentoring undergraduate and graduate students at the interface of biomolecular engineering, chemistry and structural biology. A collaboration with the science department at a large public high school with a diverse student population is also planned. These efforts will develop a workforce to support a vibrant bioeconomy.The tools of molecular dynamics simulation and x-ray crystallography will guide the directed evolution of two promising nicotinamide-dependent oxidoreductases. The focus is on engineering these enzymes for reactions in which racemic substrates can be effectively deracemized and converted to primarily one of four possible stereoisomers through dynamic reductive kinetic resolution. The targeted transformations will provide access to a spectrum of valuable building blocks for natural products synthesis, chemical biology and medicinal chemistry. Several building blocks to be constructed are for chemotherapeutic agents of the taxane family, useful as anti-cancer drugs. In previous work, the team has shown that the Clostridial enzyme (CaADH) shows remarkable substrate promiscuity, while at the same time exhibits considerable stereochemical fidelity. Preliminary results suggest that the origins of this behavior lie in a flexible binding pocket. A key goal of this project is to understand how this flexible active site is able to adapt to substrates of different shapes and functionalities and stereoselectively transform these substrates. More specifically, the team seeks to elucidate the mechanisms by which such biocatalysts effectively make two ‘stereochemical choices’, namely enantiodiscrimination (choosing one of two rapidly equilibrating enantiomers-sets first stereocenter) and facial discrimination (delivering a nicotinamide-derived hydride equivalent to one of two diastereofaces-sets second stereocenter).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

大多数药物和农业化学物质是被称为优质化学物质的。这些分子具有非常具体的结构，可以使它们具有有益的特性。酶在产生此类结构方面非常有效。该项目的目的是制定一项策略，以设计和调整参与一类称为紫杉烷的精细化学物质的酶的活性。这些化合物具有抗癌特性。目前，其中一种紫杉烷紫杉醇用于治疗转移性乳腺癌。该项目将包括在生物分子工程，化学和结构生物学界面上教育和心理化的本科生和研究生。还计划在一所大型公立高中与科学系合作，学生人数多样化。这些努力将开发一支劳动力以支持充满活力的生物经济性。分子动力学模拟和X射线晶体学的工具将指导两个承诺的烟酰胺依赖性氧化氧化激酶的定向演变。重点是工程上这些酶进行反应，在这种反应中，可以通过动态减少动力学分辨率来有效地将外星底物被有效地化并转化为四个可能的立体异构体之一。有针对性的转换将为天然产品合成，化学生物学和医学化学的范围提供一系列有价值的构件。要构建的几个构件是用于紫杉烷家族的化学治疗剂，可作为抗癌药物。在先前的工作中，该小组表明，梭状芽胞杆菌（Caadh）表现出显着的底物滥交，而同时表现出相当大的立体化学保真度。初步结果表明，这种行为的起源在于柔性结合口袋。该项目的一个关键目标是了解该灵活的活动站点如何能够适应不同形状和功能的底物，并立体选择性地改变这些基板。更具体地说，该团队试图阐明这种生物催化剂有效地做出两个“立体化学选择”的机制，即对映异构歧视（选择两个快速等效的对映异构体 - 首先立体描述器之一），并提供一个面部歧视（提供尼古丁胺 - 氢化元素等分线，以供尼古丁胺 - 降水层脱位，立体中心）。该奖项反映了NSF的法定使命，并通过使用基金会的知识分子优点和更广泛的影响标准来评估，以诚实的支持。