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筛选具有过氧化氢酶和过氧化物酶活性的污染土壤细菌分离物及其对氧化应激反应的多样性。

Screening of bacterial isolates from polluted soils exhibiting catalase and peroxidase activity and diversity of their responses to oxidative stress.

机构信息

Institute of Molecular Biology, Slovak Academy of Sciences, Dúbravská cesta 21, 84551 Bratislava, Slovakia.

出版信息

Curr Microbiol. 2010 Oct;61(4):241-7. doi: 10.1007/s00284-010-9601-x. Epub 2010 Feb 10.

Abstract

For the survival of individual isolates of gram-negative bacteria Pseudomonas putida, Achromobacter xylosoxidans, and the gram-positive bacterium Bacillus megaterium, in an environment polluted with crude oil products, the production of catalases exhibiting both catalase and dianisidine-peroxidase activity is important. Electrophoretic resolution of cell-free extracts of aerobically grown strains in Luria-Bertani medium during exponential phase revealed distinctive expression of catalatic and peroxidatic activities detected with 3,3'-diaminobenzidine tetrahydrochloride. A considerable diversity in microbial catalase and peroxidase responses to 20 or 40 mM H(2)O(2) stress, resulted from hydroperoxidase's variant of original isolates, indicating an environmental selective pressure. However, catalase was important for the adaptation of cultures to high concentration of 60 mM H(2)O(2). Appreciable differences in the sensitivity to toxic effect of H(2)O(2) (20 or 40 mM) treatment between individual isolates and their adapted variants during growth were observed until the middle of exponential phase, but they were insignificant at the entry to stationary phase. Isolates also exhibited a considerable diversity in catalases responses to phenolic contaminants 1 and 2 mM o- or p-phenylenediamine. Catalase activity of bacterium P. putida was visibly stimulated only by p-phenylenediamine and not by its positional isomer o-PDA. This study contributes to a better understanding of the role catalases play in bacterial responses to a polluted environment.

摘要

为了在受原油产品污染的环境中使革兰氏阴性菌假单胞菌、木糖氧化无色杆菌和革兰氏阳性菌巨大芽孢杆菌的个体分离物得以生存,产生同时具有过氧化氢酶和二氨基联苯胺过氧化物酶活性的过氧化氢酶是很重要的。在有氧生长的菌株的细胞提取物在 Luria-Bertani 培养基中处于指数生长期时进行电泳分离,用 3,3'-二氨基联苯胺四盐酸盐检测到了独特的过氧化氢酶和过氧化物酶活性的表达。微生物过氧化氢酶和过氧化物酶对 20 或 40mM H₂O₂应激的反应存在很大的差异,这是由于原始分离株的过氧化物酶的变体,表明存在环境选择压力。然而,过氧化氢酶对于培养物适应高浓度 60mM H₂O₂至关重要。在指数生长期的中期之前,观察到了个体分离株及其适应变体在生长过程中对 20 或 40mM H₂O₂(过氧化氢)处理的毒性效应的敏感性存在显著差异,但在进入稳定期时则不显著。分离株对 1 和 2mM 邻苯二胺或对苯二胺等酚类污染物的过氧化氢酶反应也表现出相当大的多样性。只有对苯二胺可明显刺激细菌假单胞菌的过氧化氢酶活性,而其对位异构体邻苯二胺(o-PDA)则不行。这项研究有助于更好地了解过氧化氢酶在细菌对污染环境的反应中所起的作用。

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