Tailoring biocatalysts by combining protein and cofactor engineering

酶是一种高效、选择性和可持续的化学催化剂替代品。然而，它们的应用往往受到自然界对这些生物催化剂的功能要求与实验室和工业过程要求之间的差异的阻碍。通过合理设计和实验室进化的蛋白质工程为定制酶来克服这些差异提供了强有力的解决方案。埃默里大学的首席研究员Stefan Lutz和Dale Edmondson正在探索新的工程策略，以调整黄素依赖的氧化还原酶的催化功能、底物特异性和对映体选择性。黄素依赖的氧化还原酶是生物技术和制药中不对称还原和氧化化学的重要酶类，以及生物修复应用。从蛋白质工程的角度来看，这类酶特别有趣的地方在于它们的功能互补性。该蛋白主要作为结合和预定向辅因子和底物的支架，而辅因子负责活性位点的化学反应。通过同时重塑蛋白质和辅因子，pi ？生成和评估多种变体的方法为探索协同效应提供了一个独特的机会，这些协同效应可以转化为酶的重新设计，并显著增强现有生物催化剂的功能多功能性、催化性能和物理化学性质。该合作研究项目为研究生和本科生提供了一个高度跨学科的培训环境，并为亚特兰大地区的高中教师提供了一个涉及新课程开发活动的推广框架。与埃默里大学科学教育中心和乔治亚州教师实习奖学金（GIFT）项目合作，将开发新的绿色化学和生物催化实践实验。ppi还将继续参加埃默里大学的暑期本科生研究经验（SURE）项目，与来自美国东南部的年轻学者合作，研究调整生物催化剂的新想法，其中许多是少数民族和代表性不足的群体的代表。

#1159434Lutz, StefanEnzymes can be highly effective, selective and sustainable alternatives to chemical catalysts. Nevertheless, their application is often hampered by discrepancies between the functional demands on these biocatalysts in nature versus requirements in laboratory and industrial processes. Protein engineering by rational design and laboratory evolution offers a powerful solution for tailoring enzymes to overcome these differences. Principle investigators Stefan Lutz and Dale Edmondson of Emory University are exploring new engineering strategies for tailoring the catalytic function, substrate specificity and enantioselectivity of flavin-dependent oxidoreductases, an important category of enzymes for asymmetric reduction and oxidation chemistry in biotechnological and pharmaceutical, as well as bioremediation applications. What makes this category of enzymes particularly interesting from a standpoint of protein engineering is their functional complementarity. The protein primarily serves as a scaffold to bind and pre-orient cofactor and substrate(s) while the cofactor is responsible for the chemistry in the active site. By simultaneously remodeling the protein and cofactor, the PIs? approach to generating and assessing many variants offers a unique opportunity to explore synergistic effects that can translate into enzyme redesign and significant enhancements of the functional versatility, catalytic performance and physicochemical properties of existing biocatalysts.The collaborative research project offers opportunity for a highly interdisciplinary training environment for graduate and undergraduate students, as well as provides a framework for outreach involving new curriculum development activities for Atlanta-area high school teachers. Working with the Center for Science Education at Emory and the Georgia Intern-Fellowship for Teachers (GIFT) program, new hands-on experiments on green chemistry and biocatalysis will be developed. The PIs will also continue to participate in Emory's summer undergraduate research experience (SURE) program, working with young scholars from across the southeastern United States, many of them representatives from minorities and underrepresented groups, on new ideas to tweak biocatalysts.