Enzymatic Hydrolysis of Organophosphate Esters
有机磷酸酯的酶水解
基本信息
- 批准号:9235651
- 负责人:
- 金额:$ 27.93万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2017
- 资助国家:美国
- 起止时间:2017-01-01 至 2020-12-31
- 项目状态:已结题
- 来源:
- 关键词:Active SitesAgricultureAmino Acid SequenceBacteriaBindingBiologicalChemical Warfare AgentsChemicalsComplementCrystallizationDetectionDrug Metabolic DetoxicationEnzyme Inhibitor DrugsEnzymesEstersEventExhibitsFlame RetardantsHouseholdHydrolysisInsecticidesIsotopesKineticsLabelLaboratoriesLibrariesLigandsMeasurementMethodsMonitorMutagenesisOrganophosphatesOxygenPesticidesPhosphoric Triester HydrolasesPhosphorusPlasticizersProceduresProdrugsPropertyPseudomonasReactionResearchResolutionSeriesShapesSite-Directed MutagenesisSomanStructureToxic effectVariantVirus DiseasesX ray diffraction analysisX-Ray Diffractioncatalystchemical reactioncombinatorialdesignenzyme mechanismexperimental studyinhibitor/antagonistmutantnerve agentnovelscreeningthree dimensional structure
项目摘要
The primary objective for the research described in this proposal is the elucidation of the
chemical reaction mechanism and three-dimensional structure of a novel phosphotriesterase recently
identified from the bacterium Spingobium sp. strain TCM1 (Sb-PTE). The high toxicity of many
organophosphate triesters has been exploited as the active component in many commercial
agricultural and household insecticides and as ultra-potent chemical warfare agents. Other, less toxic,
organophosphate compounds have been widely utilized as flame retardants, plasticizers, and as
prodrugs for viral infections. The catalytic properties and three-dimensional structure of the wild-
type Sb-PTE are significantly different from those exhibited by the phosphotriesterase from
Pseudomonas diminuta (Pd-PTE) or any other enzyme identified to date. We propose to utilize this
enzyme as a template for the design and creation of new biological catalysts that can be exploited for
the detection, destruction, and detoxification of toxic organophosphate nerve agents that are currently
being used as agricultural and household insecticides, plasticizers, and chemical warfare agents. The
chemical mechanism for this enzyme will be elucidated by determining the stereochemical course of
the reaction with chiral substrates and by monitoring the fate of 18-oxygen labels in the substrate and
enzyme. These experiments will be complemented by measurement of heavy atom isotope effects,
determination of the substrate/activity profile, and the identification of potent enzyme inhibitors. In
preliminary experiments we have succeeded in the crystallization of Sb-PTE and determination of its
three dimensional structure by X-ray diffraction methods. Further structures will be pursued in a
focused attempt to determine the mode of substrate binding within the active site of this enzyme.
These structures will be utilized as a guide for the design and creation of novel enzyme variants with
unique substrate profiles. Rational and combinatorial libraries of mutant enzymes will be constructed
and those variants with enhanced catalytic proficiency for the hydrolysis of toxic organophosphates
will be identified through unique screening and selection procedures. The changes in the amino acid
sequence of the Sb-PTE mutants will be correlated with enhancements in the catalytic properties and
alterations in the structure within the active site.
这项建议中描述的研究的主要目标是阐明
一种新的磷酸三酯酶的化学反应机理和三维结构
从Spingobium sp.中鉴定。菌株TCM1(SB-PTE)。许多毒物的高毒性
有机磷酸三酯已被开发为许多商业用途的活性成分。
农业和家用杀虫剂,以及作为超强化学战剂。其他,毒性较小的,
有机磷化合物已被广泛用作阻燃剂、增塑剂和
治疗病毒感染的前药。野生型的催化性能和三维结构
SB-PTE型与来自中国的磷酸三酯酶表现出的显著不同
小型假单胞菌(PD-PTE)或迄今已鉴定的任何其他酶。我们建议利用这一点
酶作为设计和创造新型生物催化剂的模板,可用于
有毒有机磷神经毒剂的检测、销毁和解毒
用作农业和家用杀虫剂、增塑剂和化学战剂。这个
该酶的化学机制将通过测定该酶的立体化学过程来阐明。
与手性底物的反应以及通过监测18-氧标记在底物中的去向和
酵素。这些实验将得到重原子同位素效应测量的补充,
底物/活性分布的测定,以及有效的酶抑制剂的鉴定。在……里面
初步实验成功地实现了Sb-PTE的结晶及其测定
用X射线衍射法测定了其三维结构。进一步的结构将在
重点尝试确定该酶活性部位内底物的结合方式。
这些结构将被用作设计和创造新型酶变体的指南
独特的基板配置文件。将构建突变酶的合理和组合文库
以及那些对有毒有机磷的水解具有更高催化熟练程度的变体
将通过独特的筛选和遴选程序进行鉴定。氨基酸的变化
Sb-Pte突变体的序列将与催化性能的增强和
活动站点内结构的更改。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Frank M. Raushel其他文献
Catalytic detoxification
催化解毒
- DOI:
10.1038/469310a - 发表时间:
2011-01-19 - 期刊:
- 影响因子:48.500
- 作者:
Frank M. Raushel - 通讯作者:
Frank M. Raushel
The use of phosphotriesterase in the synthesis of enantiomerically pure ProTide prodrugs
磷酸三酯酶在合成对映体纯的前药ProTide中的应用
- DOI:
10.1016/j.cbi.2025.111597 - 发表时间:
2025-09-05 - 期刊:
- 影响因子:5.400
- 作者:
Andrew N. Bigley;Frank M. Raushel - 通讯作者:
Frank M. Raushel
Frank M. Raushel的其他文献
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{{ truncateString('Frank M. Raushel', 18)}}的其他基金
The Discovery of Novel Metabolic Pathways for the Biosynthesis and Degradation of Complex Carbohydrates within the Human Gut Microbiome
人类肠道微生物组内复杂碳水化合物生物合成和降解的新代谢途径的发现
- 批准号:
10323657 - 财政年份:2021
- 资助金额:
$ 27.93万 - 项目类别:
The Discovery of Novel Metabolic Pathways for the Biosynthesis and Degradation of Complex Carbohydrates within the Human Gut Microbiome
人类肠道微生物组内复杂碳水化合物生物合成和降解的新代谢途径的发现
- 批准号:
10557076 - 财政年份:2021
- 资助金额:
$ 27.93万 - 项目类别:
The Discovery of Novel Metabolic Pathways for the Biosynthesis and Degradation of Complex Carbohydrates within the Human Gut Microbiome
人类肠道微生物组内复杂碳水化合物生物合成和降解的新代谢途径的发现
- 批准号:
10084621 - 财政年份:2021
- 资助金额:
$ 27.93万 - 项目类别:
Novel Biochemical Pathways for the Metabolism of Carbohydrates in the Human gut Micriobiome
人类肠道微生物组中碳水化合物代谢的新生化途径
- 批准号:
10063528 - 财政年份:2017
- 资助金额:
$ 27.93万 - 项目类别:
Deciphering Enzyme Specificity: Amidohydrolase Superfamily
破译酶的特异性:酰胺水解酶超家族
- 批准号:
7743893 - 财政年份:2009
- 资助金额:
$ 27.93万 - 项目类别:
Enzymic Detoxification for Organophosphate Nerve Agents
有机磷神经毒剂的酶解毒
- 批准号:
8114985 - 财政年份:2003
- 资助金额:
$ 27.93万 - 项目类别:
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