EAGER: Novel biocatalysts based on artificial metallo-cofactors in flavoproteins

EAGER：基于黄素蛋白中人工金属辅因子的新型生物催化剂

• 批准号: 1546790
• 负责人: Stefan Lutz
• 金额: $ 11.77万
• 依托单位: Emory University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1546790&HistoricalAwards=false
• 关键词: EAGER; Novel; biocatalysts; based; artificial

Lutz (1546790)The proposed work will generate proof-of-concept data on re-engineering the active site of the flavoprotein, Old Yellow Enzyme (OYE), to perform hydrocarbon chemistry aimed at producing chemicals that are difficult to synthesize by non-biological approaches. To this end, the proposal will focus on creating new artificial biocatalysts by integrating catalytically active organometallic complexes in the active cofactor site in the interior of the enzyme. The new biocatalysts so-formed will be assessed for their potential to catalyze C-C bond forming reactions. Moreover, the general approach of substituting artificial prosthetic co-factors has potential for general applicability to many areas of chemical synthesis and chemical conversion to useful products. The project will also feature educational components to train students in the diverse engineering and scientific disciplines required for success in the bioengineering area, and will continue the principal investigator's (PI's) involvement in a number of programs aimed at increasing interest and education in STEM areas amongst underrepresented minorities in the greater Atlanta area. The proposal capitalizes on the utility of flavin-dependent oxidoreductases as enzymes capable of biocatalysis at the laboratory scale. Specifically, the PI and his students will create new artificial biocatalysts by integrating catalytically efficient copper-bis(oxazoline)(Cu-BOX) complexes in the active site of Old Yellow Enzyme. Although challenging, the PI is well positioned to carry out this synthesis successfully based on more than three years of work in his lab on OYE which has already demonstrated substantial improvements in the enzyme's native redox activity. These studies have confirmed the PI's capability to make substantial changes in the active site without compromising the overall structural integrity of the enzyme. The modified enzymes will then be tested for catalytic activity via asymmetric cyclopropanation of styrene and ethyl diazoacetate, a benchmark reaction for C-C bond formation. By exploring new possibilities for the flavoprotein, the incorporation of artificial cofactors based on organo-metallic ligands could dramatically expand the toolbox for biocatalysis.

Lutz（1546790）拟议的工作将产生关于重新设计黄素蛋白，老黄酶（OYE）的活性位点的概念验证数据，以进行碳氢化合物化学，旨在生产难以通过非生物方法合成的化学品。 为此，该提案将集中于通过在酶内部的活性辅因子位点中整合催化活性有机金属络合物来创建新的人工生物催化剂。将评估如此形成的新生物催化剂催化C-C键形成反应的潜力。此外，替代人工假体辅因子的一般方法具有普遍适用于化学合成和化学转化为有用产物的许多领域的潜力。 该项目还将包括教育部分，以培养学生在生物工程领域取得成功所需的各种工程和科学学科，并将继续主要研究者（PI）参与一些旨在提高兴趣和教育的计划在大亚特兰大地区代表性不足的少数民族之间的STEM领域。该提案利用了黄素依赖性氧化还原酶作为能够在实验室规模上进行生物催化的酶的效用。 具体来说，PI和他的学生将通过在Old Yellow Enzyme的活性位点中整合催化效率高的铜-双（恶唑啉）（Cu-BOX）络合物来创造新的人工生物催化剂。虽然具有挑战性，PI是很好的定位，以进行这种合成成功的基础上，在他的实验室超过三年的工作对OYE已经证明了显着改善酶的天然氧化还原活性。这些研究证实了PI在活性位点进行实质性改变而不损害酶的整体结构完整性的能力。然后将通过苯乙烯和重氮乙酸乙酯的不对称环丙烷化来测试修饰的酶的催化活性，这是C-C键形成的基准反应。 通过探索黄素蛋白的新可能性，基于有机金属配体的人工辅助因子的掺入可以大大扩展生物催化的工具箱。