权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanism of a new flavin-derived cofactor involved in enzymatic decarboxylation reactions

一种新的黄素衍生辅助因子参与酶促脱羧反应的机制

基本信息

批准号：
1608553
负责人：
E. Neil Marsh
金额：
$ 50.35万
依托单位：
Regents of the University of Michigan - Ann Arbor
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1608553&HistoricalAwards=false
关键词：
Mechanism new flavin derived cofactor

项目摘要

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Neil Marsh from the University of Michigan for studies on a potentially useful new enzyme. Enzymes are proteins that speed-up many chemical reactions within cells and thus, enzymes are necessary for life. There is increasing interest in using enzymes to catalyze industrial chemical reactions because they are extremely efficient, renewable, biodegradable and non-toxic. One class of industrially promising enzymes is known as decarboxylases. These enzymes have the ability to add carbon dioxide to (or remove carbon dioxide from) molecules, which may allow them to be used in processes to develop new biofuels or important commodity chemicals such as styrene (used in making plastics). This project is studing the biochemical properties of a newly discovered decarboxylase enzyme with the long-term objective of producing this enzyme for industrial applications. In synergy with the scientific goals, the project is advancing the education, training and professional development of undergraduates, graduate students and postdoctoral scientists in the inter-disciplinary area of chemical biology. Students and postdocs are receiving training in a wide variety of specialized techniques ranging from organic synthesis to sophisticated spectroscopy experiments. The project investigates the mechanism of a novel 2-component decarboxylase system recently identified from yeast. Ferulic acid decarboxylase (FDC) catalyzes the decarboxylation of a wide range of substituted phenylacrylic acids and sorbic acid. The enzyme uses a novel, modified flavin mononucleotide (FMN) as a cofactor that contains an additional 6-membered ring derived from dimethylallyl phosphate. The second enzyme, termed PAD1, synthesizes the cofactor from reduced FMN and dimethylallyl phosphate. In particular, the Marsh group aims to determine the mechanism of FDC and role of the novel, modified flavin cofactor in catalyzing the decarboxylation reaction. They further aim to determine the mechanism by which PAD1 synthesizes the modified flavin cofactor. The experimental approaches includes measuring the kinetics of the FDC- and PAD1-catalyzed reactions using both steady state and pre-steady state techniques with a variety of substrate and coenzyme analogs. Deuterium, and 14C and 13C kinetic isotope effects are measured for FDC and combined with computational studies to fit the experimentally-determined kinetic isotope effects with models of the enzyme transition state.

有了这个奖项，化学部的生命过程化学项目将资助密歇根大学的尼尔·马什博士研究一种可能有用的新酶。酶是加速细胞内许多化学反应的蛋白质，因此，酶是生命所必需的。人们对利用酶来催化工业化学反应越来越感兴趣，因为它们非常高效、可再生、可生物降解和无毒。有一类在工业上很有前途的酶被称为脱羧酶。这些酶具有向分子中添加二氧化碳（或从分子中去除二氧化碳）的能力，这可能使它们能够用于开发新的生物燃料或重要的商品化学品，如苯乙烯（用于制造塑料）。该项目正在研究一种新发现的脱羧酶的生化特性，其长期目标是生产这种酶用于工业应用。在科学目标的协同作用下，该项目正在推进化学生物学跨学科领域的本科生、研究生和博士后科学家的教育、培训和专业发展。学生和博士后正在接受各种专业技术的培训，从有机合成到复杂的光谱实验。该项目研究了最近从酵母中鉴定出的一种新的双组分脱羧酶系统的机制。阿魏酸脱羧酶（FDC）催化一系列取代苯丙烯酸和山梨酸的脱羧反应。该酶使用一种新型的，修饰的黄素单核苷酸（FMN）作为辅助因子，该辅助因子含有来自磷酸二甲基烯丙基的额外6元环。第二种酶称为PAD1，由还原的FMN和磷酸二甲基烯丙基合成辅因子。特别是，Marsh小组旨在确定FDC的机制以及新型修饰的黄素辅助因子在催化脱羧反应中的作用。他们进一步旨在确定PAD1合成修饰的黄素辅助因子的机制。实验方法包括测量FDC-和pad1催化反应的动力学，使用稳态和预稳态技术，使用各种底物和辅酶类似物。FDC测量了氘、14C和13C的动力学同位素效应，并结合计算研究将实验确定的动力学同位素效应与酶过渡态模型拟合。