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限制硝酸单胞菌降解脂肪族氯代烃的因素:氨单加氧酶的共代谢失活和底物特异性。

Factors Limiting Aliphatic Chlorocarbon Degradation by Nitrosomonas europaea: Cometabolic Inactivation of Ammonia Monooxygenase and Substrate Specificity.

机构信息

Laboratory for Nitrogen Fixation Research, Department of Botany and Plant Pathology, 2082 Cordley Hall, Oregon State University, Corvallis, Oregon 97331-2902.

出版信息

Appl Environ Microbiol. 1991 Oct;57(10):2986-94. doi: 10.1128/aem.57.10.2986-2994.1991.

DOI:10.1128/aem.57.10.2986-2994.1991
PMID:16348568
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC183909/
Abstract

The soil nitrifying bacterium Nitrosomonas europaea is capable of degrading trichloroethylene (TCE) and other halogenated hydrocarbons. TCE cometabolism by N. europaea resulted in an irreversible loss of TCE biodegradative capacity, ammonia-oxidizing activity, and ammonia-dependent O(2) uptake by the cells. Inactivation was not observed in the presence of allylthiourea, a specific inhibitor of the enzyme ammonia monooxygenase, or under anaerobic conditions, indicating that the TCE-mediated inactivation required ammonia monooxygenase activity. When N. europaea cells were incubated with [C]TCE under conditions which allowed turnover of ammonia monooxygenase, a number of cellular proteins were covalently labeled with C. Treatment of cells with allylthiourea or acetylene prior to incubation with [C]TCE prevented incorporation of C into proteins. The ammonia-oxidizing activity of cells inactivated in the presence of TCE could be recovered through a process requiring de novo protein synthesis. In addition to TCE, a series of chlorinated methanes, ethanes, and other ethylenes were screened as substrates for ammonia monooxygenase and for their ability to inactivate the ammonia-oxidizing system of N. europaea. The chlorocarbons could be divided into three classes depending on their biodegradability and inactivating potential: (i) compounds which were not biodegradable by N. europaea and which had no toxic effect on the cells; (ii) compounds which were cooxidized by N. europaea and had little or no toxic effect on the cells; and (iii) compounds which were cooxidized and produced a turnover-dependent inactivation of ammonia oxidation by N. europaea.

摘要

土壤硝化细菌Nitrosomonas europaea 能够降解三氯乙烯 (TCE) 和其他卤代烃。N. europaea 共代谢三氯乙烯会导致 TCE 生物降解能力、氨氧化活性和细胞依赖氨的 O(2)摄取不可逆丧失。在存在氨单加氧酶的特异性抑制剂丙烯硫脲或在厌氧条件下未观察到失活,表明 TCE 介导的失活需要氨单加氧酶活性。当 N. europaea 细胞在允许氨单加氧酶周转的条件下与 [C]TCE 孵育时,许多细胞蛋白被共价标记。在用 [C]TCE 孵育之前用丙烯硫脲或乙炔处理细胞可防止 C 掺入蛋白质。在 TCE 存在下失活的细胞的氨氧化活性可以通过需要从头合成蛋白质的过程恢复。除 TCE 外,一系列氯化甲烷、乙烷和其他乙烯还被筛选为氨单加氧酶的底物以及它们使 N. europaea 的氨氧化系统失活的能力。根据它们的生物降解性和失活潜力,氯碳化合物可分为三类:(i) 不能被 N. europaea 生物降解且对细胞无毒性的化合物;(ii) 可被 N. europaea 共氧化且对细胞几乎没有毒性的化合物;和 (iii) 可被共氧化且产生依赖周转率的 N. europaea 氨氧化失活的化合物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc96/183909/0fccdd4ab54f/aem00063-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc96/183909/0fccdd4ab54f/aem00063-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc96/183909/0fccdd4ab54f/aem00063-0218-a.jpg

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Interaction of Ammonia Monooxygenase from Nitrosomonas europaea with Alkanes, Alkenes, and Alkynes.
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