A polyomic approach to elucidate the fluoranthene-degradative pathway in Mycobacterium vanbaalenii PYR-1
A polyomic approach to elucidate the fluoranthene-degradative pathway in Mycobacterium vanbaalenii PYR-1
复制标题
DOI：
10.1128/jb.00128-07
复制
发表时间：
2007-07-01
影响因子：
3.2
通讯作者：
Cerniglia, Carl E.
中科院分区：
文献类型：
--
作者：
Kweon, Ohgew;Kim, Seong-Jae;Cerniglia, Carl E.
范氏分枝杆菌PYR-1能够降解包括荧蒽在内的多种高分子量多环芳烃（PAHs）。我们使用了代谢组学，基因组学和蛋白质组学技术的组合，以调查在这种菌株的荧蒽降解。从培养基中分离出37种荧蒽代谢产物，包括潜在的异构体，并通过高效液相色谱法、气相色谱-质谱法和LN-可见光吸收法进行分析。采用液相色谱-串联质谱法结合M。vanbaalenii PYR-1基因组序列（http：jgi.doe.gov），其导致鉴定了1，122种蛋白质。其中，53种酶被确定为可能参与荧蒽降解。我们将代谢信息与基因组和蛋白质组学结果相结合，提出了荧蒽降解的途径。根据我们的假设，荧蒽的氧化是由在C-1，2，C-2，3和C-7，8位的双氧化引发的。C-1，2和C-2，3二氧基。氢化途径通过芴类代谢物降解荧蒽，而C-7，8途径通过苊烯类代谢物氧化荧蒽。双氧化的主要位点是C-2，3双氧化途径，其由18个酶促步骤组成，通过9-芴酮-1-羧酸和邻苯二甲酸酯与起始环羟基化加氧酶NidA 3B 3，将荧蒽氧化为荧蒽顺式-2，3-二氢二醇。荧蒽的非特异性单氧化与随后的二羟基荧蒽的0甲基化也作为解毒反应发生。
Mycobacterium vanbaalenii PYR-1 is capable of degrading a wide range of high-molecular-weight polycyclic aromatic hydrocarbons (PAHs), including fluoranthene. We used a combination of metabolomic, genomic, and proteomic technologies to investigate fluoranthene degradation in this strain. Thirty-seven fluoranthene metabolites including potential isomers were isolated from the culture medium and analyzed by high-performance liquid chromatography, gas chromatography-mass spectrometry, and LN-visible absorption. Total proteins were separated by one-dimensional gel and analyzed by liquid chromatography-tandem mass spectrometry in conjunction with the M. vanbaalenii PYR-1 genome sequence (http://jgi.doe.gov), which resulted in the identification of 1,122 proteins. Among them, 53 enzymes were determined to be likely involved in fluoranthene degradation. We integrated the metabolic information with the genomic and proteomic results and proposed pathways for the degradation of fluoranthene. According to our hypothesis, the oxidation of fluoranthene is initiated by dioxygenation at the C-1,2, C-2,3, and C-7,8 positions. The C-1,2 and C-2,3 dioxy. genation routes degrade fluoranthene via fluorene-type metabolites, whereas the C-7,8 routes oxidize fluoranthene via acenaphthylene-type metabolites. The major site of dioxygenation is the C-2,3 dioxygenation route, which consists of 18 enzymatic steps via 9-fluorenone-1-carboxylic acid and phthalate with the initial ring-hydroxylating oxygenase, NidA3B3, oxidizing fluoranthene to fluoranthene cis-2,3-dihydrodiol. Nonspecific monooxygenation of fluoranthene with subsequent 0 methylation of dihydroxyfluoranthene also occurs as a detoxification reaction.