Research Starter Grant

来自芽孢杆菌（Bacillus subtilus）的漆酶CotA属于多铜氧化酶家族。由于漆酶是这个家族中最简单的代表，它们被用作结构/功能关系的模型系统。CotA是一种“多面手”，一种具有广泛底物特异性的酶，它将通过实验室进化而成为一种“专家”，一种具有狭窄底物特异性的酶。揭示酶对不同底物获得特异性所必需的分子细节具有重要的智力价值。从这个项目中发现的酶应该提供可以应用于其他酶的蛋白质工程原理。底物特异性漆酶在生物传感器、生物修复和环境清洁催化剂等方面的应用前景广阔。最后，分析酶谱系的结构、动力学和光谱特性可以阐明酶如何适应其新功能。更广泛的影响：该项目有许多方面致力于促进教与学。研究生和博士后研究人员将在分子进化的新兴领域进行培训。本科生将有机会通过NJIT的Ronald E. McNair项目贡献原创研究。该项目旨在帮助低收入和第一代或代表性不足的学生学习科学、工程和数学。研究结果将在国内和国际会议上发表。此外，由于项目是多学科的，结果将在重点不同的会议上提出。这项研究也将通过新泽西理工大学预科项目中心向非技术观众进行介绍。与一名高中教师一起，首席研究员将开发一个交互式演示文稿，描述将被纳入高中课程的研究。此外，计划中的研究还能加强国际和部门内部的伙伴关系。这些有趣变体的晶体结构将由他与葡萄牙奥伊拉斯市新里斯本大学的Ligia O. Martins博士合作解决。此外，能源最小化的计算将与NJIT的Carol Venanzi教授一起完成。

The laccase CotA from Bacillus subtilus belongs to the family of multi-copper oxidase enzymes. Since laccases are the simplest representative of this family, they are used as a model system for structure/function relationships. CotA is a "generalist", an enzyme with broad substrate specificity, and it will be engineered by laboratory evolution to function as a "specialist", an enzyme with narrowed substrate specificity. Revealing the molecular details necessary for the enzyme to gain specificity toward different substrates has significant intellectual merit. Enzymes discovered from this project should provide protein engineering principles that can be applied to other enzymes. Substrate specific laccases can be further developed for applications in biosensors, bioremediation, and environmentally clean catalysts. Finally, analyzing the structures, kinetics, and spectroscopic properties of the enzyme's lineage can elucidate how the enzyme adapts towards its new function. Broader impact: The project has many aspects devoted to promoting teaching and learning. Graduate students and postdoctoral researchers will be trained in the emerging field of molecular evolution. Undergraduates will have an opportunity to contribute original research through the Ronald E. McNair program at NJIT. This program is designed to help low-income and first-generation or underrepresented students in science, engineering and mathematics. The research results will be presented at national and international meetings. Furthermore, since the project is multidisciplinary, the results will be presented at meetings that vary in focus. The research will also be described to a non-technical audience through the Center of Pre-College Programs at NJIT. Together with a high school instructor, the principal investigator will develop an interactive presentation describing the research which would be incorporated into the high school curriculum. Also, the planned research enhances international and intradepartmental partnerships. The crystal structures of the interesting variants will be solved in collaboration with Dr. Ligia O. Martins at the Universidade Nova de Lisboa in Oeiras, Portugal. Furthermore, energy minimization calculations will be done with Prof. Carol Venanzi at NJIT.