Mechanistic studies on bioremediation metalloenzymes

金属酶生物修复机理研究

基本信息

批准号：
8957328
负责人：
FERMAN Albert CHAVEZ
金额：
$ 33.42万
依托单位：
OAKLAND UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8957328
关键词：
Abate Accidents Active Sites Aerobic Amino Acid Motifs Area Aromatic Compounds Aromatic Hydrocarbons Bacteria Biological Biological Assay Biological Models Biomimetics Bioremediations Burn injury Carbon Cleaved cell Complex Cysteine dioxygenase Data Deposition Developed Countries Dioxygenases Electronics Environment Enzymes Family Fossil Fuels Gentisates Geometry Goals Health Human Hydrocarbons Hydroxylation Industry Iron Iron Compounds Life Ligands Metal Binding Site Metals Methodology Methods Modeling Mutation Nitrogen Organic Iron Compounds Organism Pesticides Petroleum Plastics Poison Property Psyche structure Publishing Qualifying Reaction Reporting Research Salicylic Acids Soil Solvents Source Structure Students Study models System Testing Tweens Waste Products Work analog carboxylate catalyst cold temperature collaborative environment enzyme mechanism extradiol dioxygenase innovation insight metal complex metalloenzyme microorganism novel oxidation physical property pollutant public health relevance rapid growth

项目摘要

DESCRIPTION (provided by applicant): Aromatic compounds are commonly formed during the incomplete burning of fossil fuels, solvents, pesticides and plastics. Their placement within the biosphere either intentionally or accidentally has been a problem for industrialized countries. Bioremediation is the intentional use and manipulation of living organisms to remove environ- mental pollutants. The combination of rapid growth rate, global abundance and high rate of mutations enables bacteria to adapt to utilize pollutants as a carbon source. In aerobic microorganisms, activation of an aromatic substrate is usually effected by hydroxylation of the ring and subsequent dearomatization. Ring-fission dioxygenases that catalyze these reactions contain Fe3+ ions (intradiol dioxygenases) or Fe2+ ions (extradiol cleaving enzymes). A third Class of dioxygenases has been recently identified. These enzymes belong to the cupin super- family, which is characterized by a six-stranded -barrel fold and conserved amino acid motifs providing 3His or 2- or 3His-1Glu ligand environments to metal ions. The enzymes gentisate 1,2-dioxygenase (GDO) and salicylate 1,2-dioxygenase (SDO) belong to this new class and contain a 3-His metal binding site. Some mechanistic work has been reported for GDO, however, essentially no mechanistic work for SDO has been published. This provides an opportunity to investigate the mechanism for SDO using model studies since it is unlikely to be the same as GDO due to structural differences in the substrates. Additionally, a scarcely characterize bioremediation enzyme (2,4'-dihydroxyacetophenone dioxygenase, DAD) capable of oxidizing 2,4'-dihydroxyacetophenone (DHAP) has been identified. This cupin enzyme contains a 3-His-1-Glu active site. Synthetic model studies can assist in elucidating the mechanism for this new enzyme as well. In this proposal we aim to synthesize iron compounds using organic ligands of varying properties. We will synthesize three nitrogen and three nitrogen-one-carboxylate ligands. These ligands will be reacted with iron(II) and iron(III) to generate model systems with tunable coordination geometry and electronic properties. The complexes will be systematically studied to generate correlations be- tween the model complex structure and spectroscopic and physical properties. Compounds which are deemed good structural and spectroscopic models for selected bioremediation enzymes will be studied for biomimetic activity. Next, we will probe the reactivity of model complexes towards aromatic ring-containing compounds. Assays will be performed to test the complexes for both stoichiometric and catalytic dioxygenase activity. If intermediates are observed at low temperature, we will attempt to isolate and characterize these species. Such information will provide mechanistic insights relevant to bioremediation enzymes thus providing a better understanding of how these new classes of enzymes performs their function.

描述（由申请人提供）：芳香族化合物是在化石燃料、溶剂、农药和塑料不完全燃烧过程中通常形成的化合物，它们有意或无意地进入生物圈一直是工业化国家的一个问题。生物修复是有意识地利用和操纵活生物体来去除环境污染物，快速生长、全球丰度和高突变率的结合使细菌能够适应利用污染物作为碳源，从而激活需氧微生物。芳香族底物通常受到环的羟基化和随后的环裂变双加氧酶的影响，催化这些反应含有 Fe3+ 离子（间二醇）。双加氧酶）或 Fe2+ 离子（extradiol 裂解酶）最近已鉴定出第三类双加氧酶，这些酶属于 cupin 超家族，其特征是六链  桶折叠和提供 3His 的保守氨基酸基序。或 2- 或 3His-1Glu 配体环境与金属离子龙胆酸 1,2-双加氧酶。 (GDO) 和水杨酸 1,2-双加氧酶 (SDO) 属于这一新类别，并含有 3-His 金属结合位点。已报道了 GDO 的一些机械工作，但是，基本上没有发表过 SDO 的机械工作。提供了使用模型研究研究 SDO 机制的机会，因为由于底物的结构差异，它不太可能与 GDO 相同。此外，这是一种几乎没有特征的生物修复酶。 (2,4'-二羟基苯乙酮双加氧酶，DAD) 已被鉴定出能够氧化 2,4'-二羟基苯乙酮 (DHAP)。这种 cupin 酶含有 3-His-1-Glu 活性位点，合成模型研究可以帮助阐明。在这个提案中，我们的目标是使用不同性质的有机配体合成铁化合物，我们将合成三个氮和三个氮一羧酸盐。这些配体将与铁（II）和铁（III）反应生成具有可调配位几何和电子性质的模型系统，将系统地研究这些配合物，以生成模型配合物结构与光谱和物理性质之间的相关性。被认为是良好的结构和光谱模型的选定的生物修复酶将进行仿生活性研究接下来，我们将探讨模型复合物对含芳香环化合物的反应性。将进行测试复合物的化学计量和催化双加氧酶活性，如果在低温下观察到中间体，我们将尝试分离和表征这些物种，这些信息将提供与生物修复酶相关的机制见解，从而更好地了解如何进行。这些新型酶发挥其功能。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Synthesis and reactivity of a 4His enzyme model complex.

4His 酶模型复合物的合成和反应性。

DOI：
发表时间：
2017
期刊：
影响因子：
3.9
作者：
Li, Jia;Banerjee, Atanu;Hasse, Timothy A;Loloee, Reza;Biros, Shannon M;Staples, Richard J;Chavez, Ferman A
通讯作者：
Chavez, Ferman A

A Biomimetic System for Studying Salicylate Dioxygenase.

用于研究水杨酸双加氧酶的仿生系统。

DOI：
发表时间：
2019-01-01
期刊：
影响因子：
0
作者：
Banerjee, Atanu;Li, Jia;Molenda, Monika A;Brennessel, William W;Chavez, Ferman A
通讯作者：
Chavez, Ferman A

Self-assembly of multiferroic core-shell composites using DNA functionalized nanoparticles.

使用 DNA 功能化纳米粒子自组装多铁性核壳复合材料。

DOI：
10.1016/j.jmmm.2018.03.075
发表时间：
2018-08-15
期刊：
Journal of magnetism and magnetic materials
影响因子：
2.7
作者：
Banerjee A;Zhang J;Zhou P;Tuppil K;Sreenivasulu G;Qu H;Zhang T;Timilsina R;Chavez FA;Srinivasan G
通讯作者：
Srinivasan G

A Structural Model for the Iron-Nitrosyl Adduct of Gentisate Dioxygenase.

龙胆酸双加氧酶铁-亚硝酰加合物的结构模型。

DOI：
10.1002/ejic.201800992
发表时间：
2018-11-27
期刊：
European journal of inorganic chemistry
影响因子：
2.3
作者：
A. Banerjee;J. Li;Amy L. Speelman;C. White;P. Pawlak;W. Brennessel;N. Lehnert;Ferman A. Chavez
通讯作者：
Ferman A. Chavez

Structural, Spectroscopic, Electrochemical, and Magnetic Properties for Manganese(II) Triazamacrocyclic Complexes.

三氮杂大环锰 (II) 配合物的结构、光谱、电化学和磁性。

DOI：
10.1016/j.ica.2018.11.013
发表时间：
2019-02-24
期刊：
Inorganica chimica acta
影响因子：
2.8
作者：
Banerjee A;Tolla AS;Stjepanovic S;Sevilla MD;Goodsell JL;Angerhofer A;Brennessel WW;Loloee R;Chavez FA
通讯作者：
Chavez FA