Mechanistic Enzymology of Bacterial Lignin Degradation

细菌木质素降解的机理酶学

基本信息

批准号：
BB/M003523/1
负责人：
Timothy Bugg
金额：
$ 68.37万
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2014
资助国家：
英国
起止时间：
2014 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FM003523%2F1
关键词：
Mechanistic Enzymology Bacterial Lignin Degradation

项目摘要

The chemicals that are used to make plastics, drugs, household chemicals & industrial solvents come primarily from oil, so one of the greatest scientific challenges for the 21st century is how we can find renewable sources for these important chemicals. One obvious place to look is plant lignocellulose, the major component of plant cell walls, found in biomass such as agricultural waste. One component of plant lignocellulose is lignin: an aromatic polymer that is naturally hard to break down, since it is held together by C-O ether linkages and C-C bonds that are not broken down by acid or base treatment. We have isolated bacteria from soil that are able to break down lignin, we have studied in detail the first bacterial lignin-degrading enzyme, an enzyme called peroxidase DypB from Rhodococcus jostii RHA1, and we are beginning to understand the biochemical pathways used by bacteria to break down lignin.We have recently identified a second type of enzyme that can break down lignin,a manganese-dependent superoxide dismutase from Sphingobacterium sp. T2, This is the first time that an enzyme of this kind has been shown to have activity towards lignin. We wish to study in detail the reaction mechanism of this enzyme, to determine exactly what is the reactive species that is responsible for attacking lignin. We have also found another bacterial dyp peroxidase from Pseudomonas fluorescens that can attack lignin, and so we wish to determine the structure of the product that it releases, and determine the mechanism of this reaction. We have also recently determined the three-dimensional structure of a Dyp peroxidase enzyme from Thermobifida fusca; we wish to use this structure to carry out "directed evolution" on this enzyme, to improve its activity towards lignin.We believe that, as well as the lignin-oxidising enzymes such as Dyp peroxidases and Mn superoxide dismutase, there must be other accessory enzymes and proteins that are needed for bacterial lignin breakdown. In this project, we will try to identify an esterase enzyme that breaks an ester linkage between lignin and hemi-cellulose; and a dehydrogenase enzyme that assists lignin breakdown by quenching radical intermediates generated during lignin breakdown. We will then test combinations of these accessory enzymes and lignin-degrading enzymes with samples of lignin either from plant biomass, or from industrial sources, to test whether they are effective for lignin breakdown in vitro, and whether we can use these enzymes to produce aromatic chemicals.

用于制造塑料，药物，家用化学物质和工业溶剂的化学物质主要来自石油，因此21世纪最大的科学挑战之一是我们如何找到这些重要化学物质的可再生能源。一个显而易见的地方是植物木质纤维素，这是植物细胞壁的主要组成部分，在农业废物等生物量中发现。植物木质纤维素的一个组成部分是木质素：一种自然很难分解的芳香族聚合物，因为它是由C-O醚链接和C-C键一起固定的，而C-C键不会被酸或碱处理分解。我们有能够分解木质素的土壤中孤立的细菌，我们详细研究了第一个细菌木质素降解酶，一种酶，一种称为jostii rha1的酶，称为过氧化物酶DYPB，我们正在开始识别lignignin的生物学途径。来自鞘杆菌的锰依赖性超氧化物歧化酶。 T2，这是第一次证明这种酶对木质素具有活性。我们希望详细研究该酶的反应机制，以确定导致攻击木质素的反应性物种是什么。我们还从假单胞菌荧光症中发现了另一个可攻击木质素的细菌DYP过氧化物酶，因此我们希望确定其释放的产物的结构，并确定该反应的机理。我们最近还确定了来自Thermobifida Fusca的DYP过氧化物酶的三维结构；我们希望使用这种结构在该酶上进行“定向进化”，以改善其对木质素的活性。我们相信，以及诸如DYP过氧化物酶和MN超氧化物歧化酶之类的木质素氧化酶，必须有其他辅助酶和蛋白质，这些酶和蛋白质是细菌木质林蛋白破裂所需的。在这个项目中，我们将尝试确定一种酯酶，该酯酶破坏木质素和半纤维素之间的酯连接。以及一种通过淬灭木质素分解过程中产生的自由基中间体来有助于木质素分解的脱氢酶。然后，我们将测试这些辅助酶和木质素降解酶与木质素样品中的木质素酶的组合，以测试它们是否可以在体外有效，以及我们是否可以使用这些酶生产芳香族化学物质。

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）

Extracellular alpha/beta-hydrolase from Paenibacillus species shares structural and functional homology to tobacco salicylic acid binding protein 2.

来自类芽孢杆菌属物种的胞外 α/β-水解酶与烟草水杨酸结合蛋白 2 具有结构和功能同源性。

DOI：
10.1016/j.jsb.2020.107496
发表时间：
2020
期刊：
Journal of structural biology
影响因子：
3
作者：
Wilkinson RC
通讯作者：
Wilkinson RC

A single sensor controls large variations in zinc quotas in a marine cyanobacterium.

DOI：
10.1038/s41589-022-01051-1
发表时间：
2022-08
期刊：
Nature chemical biology
影响因子：
14.8
作者：
Mikhaylina A;Ksibe AZ;Wilkinson RC;Smith D;Marks E;Coverdale JPC;Fülöp V;Scanlan DJ;Blindauer CA
通讯作者：
Blindauer CA

Characterization of Thiamine Diphosphate-Dependent 4-Hydroxybenzoylformate Decarboxylase Enzymes from Rhodococcus jostii RHA1 and Pseudomonas fluorescens Pf-5 Involved in Degradation of Aryl C2 Lignin Degradation Fragments.

红球菌 jostii RHA1 和荧光假单胞菌 Pf-5 参与芳基 C2 木质素降解片段降解的硫胺素二磷酸依赖性 4-羟基苯甲酰甲酸脱羧酶的表征。

DOI：
10.1021/acs.biochem.9b00177
发表时间：
2019
期刊：
Biochemistry
影响因子：
2.9
作者：
Wei Z
通讯作者：
Wei Z

Sphingobacterium sp. T2 Manganese Superoxide Dismutase Catalyzes the Oxidative Demethylation of Polymeric Lignin via Generation of Hydroxyl Radical.

DOI：
10.1021/acschembio.8b00557
发表时间：
2018-09
期刊：
ACS chemical biology
影响因子：
4
作者：
G. M. Rashid;Xiaoyang Zhang;R. Wilkinson;V. Fülöp;B. Cottyn;S. Baumberger;T. Bugg
通讯作者：
G. M. Rashid;Xiaoyang Zhang;R. Wilkinson;V. Fülöp;B. Cottyn;S. Baumberger;T. Bugg

Identification of Manganese Superoxide Dismutase from Sphingobacterium sp. T2 as a Novel Bacterial Enzyme for Lignin Oxidation.

DOI：
10.1021/acschembio.5b00298
发表时间：
2015-08
期刊：
ACS chemical biology
影响因子：
4
作者：
G. M. Rashid;Charles R. Taylor;Yangqingxue Liu;Xiaoyang Zhang;D. Rea;V. Fülöp;T. Bugg
通讯作者：
G. M. Rashid;Charles R. Taylor;Yangqingxue Liu;Xiaoyang Zhang;D. Rea;V. Fülöp;T. Bugg