Quantification of Naphthalene Dioxygenase (NahAC) and Catechol Dioxygenase (C23O) Catabolic Genes Produced by Phenanthrene-Degrading Pseudomonas fluorescens AH-40,Current Genomics

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Quantification of Naphthalene Dioxygenase (NahAC) and Catechol Dioxygenase (C23O) Catabolic Genes Produced by Phenanthrene-Degrading Pseudomonas fluorescens AH-40
Current Genomics ( IF 1.8 ) Pub Date : 2020-05-20 , DOI: 10.2174/1389202921666200224101742
Asmaa M M Mawad ₁ , Wael S Abdel-Mageed ₁ , Abd E-L Hesham ₁

Affiliation

Background Petroleum polycyclic aromatic hydrocarbons (PAHs) are known to be toxic and carcinogenic for humans and their contamination of soils and water is of great environmental concern. Identification of the key microorganisms that play a role in pollutant degradation processes is relevant to the development of optimal in situ bioremediation strategies. Objective Detection of the ability of Pseudomonas fluorescens AH-40 to consume phenanthrene as a sole carbon source and determining the variation in the concentration of both nahAC and C23O catabolic genes during 15 days of the incubation period. Methods In the current study, a bacterial strain AH-40 was isolated from crude oil polluted soil by enrichment technique in mineral basal salts (MBS) medium supplemented with phenanthrene (PAH) as a sole carbon and energy source. The isolated strain was genetically identified based on 16S rDNA sequence analysis. The degradation of PAHs by this strain was confirmed by HPLC analysis. The detection and quantification of naphthalene dioxygenase (nahAc) and catechol 2,3-dioxygenase (C23O) genes, which play a critical role during the mineralization of PAHs in the liquid bacterial culture were achieved by quantitative PCR. Results Strain AH-40 was identified as pseudomonas fluorescens. It degraded 97% of 150 mg phenanthrene L-1 within 15 days, which is faster than previously reported pure cultures. The copy numbers of chromosomal encoding catabolic genes nahAc and C23O increased during the process of phenanthrene degradation. Conclusion nahAc and C23O genes are the main marker genes for phenanthrene degradation by strain AH-40. P. fluorescence AH-40 could be recommended for bioremediation of phenanthrene contaminated site.

中文翻译：

菲降解荧光假单胞菌 AH-40 产生的萘双加氧酶 (NahAC) 和儿茶酚双加氧酶 (C23O) 分解代谢基因的定量

背景众所周知，石油多环芳烃 (PAH) 对人类有毒和致癌，并且它们对土壤和水的污染引起了极大的环境关注。确定在污染物降解过程中发挥作用的关键微生物与开发最佳原位生物修复策略有关。目的检测荧光假单胞菌AH-40消耗菲作为唯一碳源的能力，并确定nahAC和C23O分解代谢基因在15天潜伏期内的浓度变化。方法在本研究中，在以菲（PAH）为唯一碳源和能源的基础矿物盐（MBS）培养基中，采用富集技术从原油污染土壤中分离出菌株AH-40。基于 16S rDNA 序列分析对分离的菌株进行遗传鉴定。通过 HPLC 分析证实了该菌株对 PAHs 的降解。通过定量 PCR 实现了对液体细菌培养物中多环芳烃矿化过程中起关键作用的萘双加氧酶 (nahAc) 和儿茶酚 2,3-双加氧酶 (C23O) 基因的检测和定量。结果菌株AH-40被鉴定为荧光假单胞菌。它在 15 天内降解了 97% 的 150 mg 菲 L-1，这比之前报道的纯培养物要快。在菲降解过程中，编码分解代谢基因nahAc和C23O的染色体拷贝数增加。结论nahAc和C23O基因是AH-40菌株降解菲的主要标志基因。P。

更新日期：2020-05-20

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