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Tendency of using different aromatic compounds as substrates by 2,4-DNT dioxygenase expressed by pJS39 carrying the gene dntA from Burkholderia sp. strain DNT
Bioremediation Journal ( IF 2 ) Pub Date : 2018-11-13 , DOI: 10.1080/10889868.2018.1516619
Khaled M. Khleifat 1 , Muhamad O. Al-limoun 1 , Khalid Y. Alsharafa 1 , Haitham Qaralleh 2 , Amjad A. Al Tarawneh 3
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Abstract The substrate range of 2,4-dinitrotoluene (DNT) dioxygenase was investigated by measuring substrate-dependent O2 uptake and maximum growth (expressed in A600) on substrate-containing minimal medium. The control for each strain had no added particular substrate. The following aromatic compounds: catechol, α-naphthalene acetic acid, β-dimethylaminobenzaldehyde, 3,4-dinitrosalicylic acid, p-nitrophenol, naphthanol, o-anisic acid, salicylic acid, toluene, and benzoic acid, were tried as possible substrates. Considering all substrates used, only p-nitrophenol showed zero oxygen uptake rate and zero growth. This indicates that it was rather unlikely that p-nitrophenol is a substrate analog for 2,4-DNT. Catechol was clearly used as a sole carbon source by both wild-type Escherichia. coli (JM103) and the dnt transformant (JS39). Using α-naphthalene acetic acid and β-dimethylaminobenzaldehyde as substrates resulted in DNT dioxygenase oxygen uptake rates of 11.8 and 14 μM/hr/mg protein, respectively. However, using both compounds as a carbon source, JS39 had twice the growth rate of E. coli JM103. For the remaining six substrates tested (3, 4-dinitrosalicylic acid, p-nitrophenol, o-anisic acid, salicylic acid, toluene, and benzoic acid), there appeared to be growth advantages for JS39 (even though the growth in the presence of substrate was less than the controls) suggesting a situation similar to that described for α-naphthalene and β-dimethylaminobenzaldehyde above. Combining results from our assay with respirometry and growth-based experiments will allow a better understanding of the biochemical consequences of these interactions. These results suggest that DNT dioxygenase gene, dntA carried by JS39, and those potential genes for substrates-degraded enzyme(s) system could have a common root.

中文翻译:

通过携带来自伯克霍尔德氏菌属的基因 dntA 的 pJS39 表达的 2,4-DNT 双加氧酶使用不同芳香族化合物作为底物的趋势。应变DNT

摘要 2,4-二硝基甲苯 (DNT) 双加氧酶的底物范围通过在含底物的基本培养基上测量底物依赖性 O2 吸收和最大生长(以 A600 表示)来研究。每个菌株的对照没有添加特定的底物。以下芳香族化合物:儿茶酚、α-萘乙酸、β-二甲氨基苯甲醛、3,4-二硝基水杨酸、对硝基苯酚、萘醇、邻茴香酸、水杨酸、甲苯和苯甲酸,被尝试作为可能的底物。考虑到所有使用的底物,只有对硝基苯酚表现出零氧摄取率和零生长。这表明对硝基苯酚不太可能是 2,4-DNT 的底物类似物。儿茶酚显然被两种野生型埃希氏菌用作唯一的碳源。大肠杆菌 (JM103) 和 dnt 转化体 (JS39)。使用 α-萘乙酸和 β-二甲氨基苯甲醛作为底物导致 DNT 双加氧酶的氧摄取率分别为 11.8 和 14 μM/hr/mg 蛋白质。然而,使用这两种化合物作为碳源,JS39 的生长速度是大肠杆菌 JM103 的两倍。对于测试的其余六种底物(3, 4-二硝基水杨酸、对硝基苯酚、邻茴香酸、水杨酸、甲苯和苯甲酸),JS39 似乎具有生长优势(即使在存在底物少于对照)表明情况类似于上述α-萘和β-二甲氨基苯甲醛的情况。将我们的测定结果与呼吸测定法和基于生长的实验相结合,可以更好地了解这些相互作用的生化后果。
更新日期:2018-11-13
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