Anaerobic Toluene Metabolism in the T1 Denitrifying Strain

;​R o o t E n t y F VxZ6 @ C o m p o b j b W o r d d o c u m e n t F c O b j e t P O O l 6 uZ6 6 uZ64 @ 6 7 @ 8 9:；= ?@ F Microsoft Word 6.0 Document MSWordDoc6;​{{{{Frazer, Anne C. MCB-9507132这个项目的目标是表征菌株T1厌氧降解甲苯的生化特征。这是在该实验室获得的一种新分离物，并且已经在微生物利用甲苯的厌氧途径上进行了初步工作。甲苯在T1中有两种命运，要么是生产性代谢并最终转化为二氧化碳和生物质，要么是转化为两种终端代谢物：琥珀酸苄酯和富马酸苄酯。在厌氧甲苯代谢过程中，在某些硫酸盐还原环境和其他几种反硝化甲苯降解剂中也报道了同样的终端产物的形成。因此，这种转变可能对环境有重大意义。提出了利用厌氧细胞提取物进行酶分析，重点是鉴定T1代谢甲苯的关键酶步骤的反应物和产物。重点包括确定甲苯利用途径中的初始酶步骤，指定TUT途径，以及确定甲苯终端代谢物形成的初始步骤。提出的研究是对实验室同事目前正在进行的关于菌株T1厌氧甲苯降解遗传特征的研究的补充，并将利用不能使用甲苯的突变体来解决甲苯降解途径。甲苯是一种主要的工业化学品，也是燃料的一种成分。它在土壤和水生环境中也是一种危险废物。关于空气中甲苯的降解，人们已经了解了很多。然而，污染场所的氧气被迅速消耗，这使得研究厌氧代谢变得非常重要。本项目的目的是表征T1厌氧代谢甲苯的初始反应。甲苯在T1中有两种命运，要么矿化，要么转化为两种终端化学产物，在生长介质中积累。首先将进行研究，确定细胞提取物中甲苯代谢的初始反应，目的是继续确定不同降解途径中的关键酶促反应物和产物。该项目将有助于更好地了解环境中的自然降解过程，并有效地利用这些过程来修复受污染的场地。*** ;Oh + 0 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $ H + 1 $R:模板\ WWUSER \ \正常。点 弗雷泽,安妮·c·罗伯特·乌芬雪莱·a·格雷夫斯@ Sv 4 @ @ tfZ6 @ Ik微软Word 6.0 2.0；​;e = e F j j j j j j j 1 # % % % C C T @ j j j这个项目的目标是

; R o o t E n t r y F VxZ6 @ C o m p O b j b W o r d D o c u m e n t F O b j e c t P o o l 6 uZ6 6 uZ6 4 @ 6 7 8 9 : ; = ? @ F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; { { { { Frazer, Anne C. MCB-9507132 The goal of this project is to characterize the biochemical features of the anaerobic degradation of toluene by strain T1. This is a novel isolate obtained in this laboratory and preliminary work has already been done on the anaerobic pathway for toluene utilization in the microbe. Toluene has two fates in T1, either productive metabolism and eventual conversion to carbon dioxide and biomass, or transformation to two dead-end metabolites, benzylsuccinate and benzylfumarate. The formation of the same dead-end products during anaerobic toluene metabolism has also recently been reported in certain sulfate-reducing environments, and in several other denitrifying toluene degraders. Thus, ths transformation may be environmentally significant. It is proposed to develop enzyme assays using anaerobic cell extracts with an emphasis on identifying the reactants and products of key enzyme steps in the toluene metabolism by T1. Emphasis includes, determining the initial enzyme step in the toluene utilization pathway, designated the TUT pathway, as well as defining the initi al step in the formation of the toluene dead-end metabolites. The research proposed is complementary to work currently underway by colleagues in the laboratory working on the genetic features of anaerobic toluene degradation by strain T1, and will make use of mutants unable to use toluene in order to resolve toluene degradation pathways. %%% Toluene is a major industrial chemical as well as a component of fuel. It is also a hazardous waste in soil and aquatic environments. A good deal has been learned about the degradation of toluene in air. However, oxygen in pollution sites is rapidly used up, which makes the study of anaerobic metabolism so important. The goal of this project is to characterize the beginning reactions performed by T1 anaerobically metabolize toluene. Toluene has two fates in T1, either mineralization or transformation into two dead-end chemical products that accumulate in the growth medium. Studies will first be undertaken to identify initial reactions of toluene metabolism in extracts from cells, with the intention of continuing on to identify key enzymatic reactants and products in the separate degradation pathways. This project will help provide a better understanding about the natural degradative processes in the environment and to use them effectively to remediate contaminated sites. *** ; Oh +' 0 $ H l D h S u m m a r y I n f o r m a t i o n ( 5 R:\WWUSER\TEMPLATE\NORMAL.DOT Frazer, Anne C. Robert Uffen Shelley A. Graves @ Sv 4 @ @ tfZ6 @ Ik Microsoft Word 6.0 2 ; ; e = e F j j j j j j j 1 # % % % C c T @ j j j j j # ~ j j j j # { Frazer, Anne C. MCB-9507132 The goal of this project is to ch