He Chunqiu, Li Yanpeng, Huang Chao, Chen Fulin, Ma Yanling
Shaanxi Provincial Key Laboratory of Biotechnology, Key Laboratory of Resources Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi'an, China.
Front Microbiol. 2018 Oct 31;9:2595. doi: 10.3389/fmicb.2018.02595. eCollection 2018.
DN1, isolated from petroleum-contaminated soil, showed excellent degradation ability toward diverse polycyclic aromatic hydrocarbons (PAHs). Many studies have been done to improve its degradation ability. However, the molecular mechanisms of PAHs degradation in DN1 strain are unclear. In this study, the whole genome of DN1 strain was sequenced and analyzed. Its genome contains 6,641,902 bp and encodes 6,684 putative open reading frames (ORFs), which has the largest genome in almost all the comparative strains. Results of gene annotation showed that this strain harbored over 100 candidate genes involved in PAHs degradation, including those encoding 25 dioxygenases, four ring-hydroxylating dioxygenases, five ring-cleaving dioxygenases, and various catabolic enzymes, transcriptional regulators, and transporters in the degradation pathways. In addition, gene knockout experiments revealed that the disruption of some key PAHs degradation genes in DN1 strain, such as A, G, H, and A, did not completely inhibit fluoranthene degradation, even though their degradative rate reduced to some extent. Three intermediate metabolites, including 9-hydroxyfluorene, 1-acenaphthenone, and 1, 8-naphthalic anhydride, were identified as the dominating intermediates in presence of 50 μg/mL fluoranthene as the sole carbon source according to gas chromatography mass spectrometry analysis. Taken together, the genomic and metabolic analysis indicated that the fluoranthene degradation by DN1 strain was initiated by dioxygenation at the C-1, 2-, C-2, 3-, and C-7, 8- positions. These results provide new insights into the genomic plasticity and environmental adaptation of DN1 strain.
从石油污染土壤中分离出的DN1对多种多环芳烃(PAHs)表现出优异的降解能力。人们已开展了许多研究来提高其降解能力。然而,DN1菌株中PAHs降解的分子机制尚不清楚。在本研究中,对DN1菌株的全基因组进行了测序和分析。其基因组包含6,641,902 bp,编码6,684个推定的开放阅读框(ORF),在几乎所有比较菌株中具有最大的基因组。基因注释结果表明,该菌株含有100多个参与PAHs降解的候选基因,包括编码25种双加氧酶、4种环羟基化双加氧酶、5种环裂解双加氧酶以及降解途径中的各种分解代谢酶、转录调节因子和转运蛋白的基因。此外,基因敲除实验表明,DN1菌株中一些关键的PAHs降解基因(如A、G、H和A)的破坏并未完全抑制荧蒽的降解,尽管其降解率在一定程度上有所降低。根据气相色谱-质谱分析,在以50 μg/mL荧蒽作为唯一碳源的情况下,三种中间代谢产物,包括9-羟基芴、1-苊酮和1,8-萘二甲酸酐,被确定为主要中间产物。综上所述,基因组和代谢分析表明,DN1菌株对荧蒽的降解是通过在C-1,2-、C-2,3-和C-7,8-位的双加氧作用启动的。这些结果为DN1菌株的基因组可塑性和环境适应性提供了新的见解。