Systematic unraveling of the unsolved pathway of nicotine degradation in Pseudomonas.
Systematic unraveling of the unsolved pathway of nicotine degradation in Pseudomonas.
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系统地揭示假单胞菌中尼古丁降解的未解途径
DOI：
10.1371/journal.pgen.1003923
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发表时间：
2013-10
期刊：
影响因子：
4.5
通讯作者：
Xu P
中科院分区：
文献类型：
--
作者：
Tang H;Wang L;Wang W;Yu H;Zhang K;Yao Y;Xu P
恶臭假单胞菌等微生物在有机废物和有毒化合物的矿化过程中起着重要作用。为了全面和准确地阐明恶臭假单胞菌尼古丁降解的关键过程，我们用基于MS的光谱计数法测量了恶臭假单胞菌S16在尼古丁或甘油(一种非抑制性碳源)上生长的差异蛋白丰度水平。在计算机分析中，突出了参与尼古丁降解的功能途径的蛋白质的显著聚集。用定量逆转录-聚合酶链式反应分析差异表达基因的转录调控。我们观察到以下关键结果：(I)包含1,292个观察到的蛋白质的蛋白质组提供了参与尼古丁代谢的酶的详细视图。这些蛋白质可以归类为运输、解毒和氨基酸代谢的功能基团。生长在尼古丁和甘油中的细胞胞浆蛋白图谱有显著差异。(Ii)3-琥珀酰基吡啶转化为6-羟基-3-琥珀酰基吡啶的关键步骤是由钼多酚结合酶(SPMA)、钼多酚脱氢酶(SpmB)和(2Fe-2S)结合铁氧还蛋白(SpmC)组成的多酶反应，其中钼多菌素胞嘧啶二核苷酸为辅因子。(3)克隆了一种新的尼古丁氧化还原酶(NicA2)基因，并对重组蛋白进行了鉴定。目前研究中确定的蛋白质和功能途径是环境有毒化合物降解的有吸引力的靶点。
Microorganisms such as Pseudomonas putida play important roles in the mineralization of organic wastes and toxic compounds. To comprehensively and accurately elucidate key processes of nicotine degradation in Pseudomonas putida, we measured differential protein abundance levels with MS-based spectral counting in P. putida S16 grown on nicotine or glycerol, a non-repressive carbon source. In silico analyses highlighted significant clustering of proteins involved in a functional pathway in nicotine degradation. The transcriptional regulation of differentially expressed genes was analyzed by using quantitative reverse transcription-PCR. We observed the following key results: (i) The proteomes, containing 1,292 observed proteins, provide a detailed view of enzymes involved in nicotine metabolism. These proteins could be assigned to the functional groups of transport, detoxification, and amino acid metabolism. There were significant differences in the cytosolic protein patterns of cells growing in a nicotine medium and those in a glycerol medium. (ii) The key step in the conversion of 3-succinoylpyridine to 6-hydroxy-3-succinoylpyridine was catalyzed by a multi-enzyme reaction consisting of a molybdopeterin binding oxidase (spmA), molybdopterin dehydrogenase (spmB), and a (2Fe-2S)-binding ferredoxin (spmC) with molybdenum molybdopterin cytosine dinucleotide as a cofactor. (iii) The gene of a novel nicotine oxidoreductase (nicA2) was cloned, and the recombinant protein was characterized. The proteins and functional pathway identified in the current study represent attractive targets for degradation of environmental toxic compounds.
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LysR 家族转录调节因子 PqsR 作为假单胞菌 M18 中脓毒素生物合成的阻遏物和吩嗪-1-羧酸生物合成的激活剂
影响因子：
4.1
作者：
Lu, Jishun;Huang, Xianqing;Xu, Yuquan
通讯作者：
Xu, Yuquan
DOI：
10.1038/nbt1183-784
发表时间：
1983-01-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
SIMON, R;PRIEFER, U;PUHLER, A
通讯作者：
PUHLER, A
影响因子：
3.9
作者：
WADA, E
通讯作者：
WADA, E
假单胞菌HZN6菌株中两个参与尼古丁分解代谢的新基因的功能鉴定
影响因子：
4.4
作者：
Qiu, Jiguo;Ma, Yun;Liu, Weiping
通讯作者：
Liu, Weiping
对恶臭假单胞菌 S16 马来酸异构酶的结构和计算研究揭示了将底物包裹在内部的呼吸运动
影响因子：
3.6
作者：
Chen, Duoduo;Tang, Hongzhi;Xu, Ping
通讯作者：
Xu, Ping