OnpA, an Unusual Flavin-Dependent Monooxygenase Containing a Cytochrome b(5) Domain
文献类型：期刊论文
| 作者 | Xiao, Yi1; Liu, Ting-Ting1; Dai, Hui2; Zhang, Jun-Jie1; Liu, Hong1; Tang, Huiru2; Leak, David J.3; Zhou, Ning-Yi1 |
| 刊名 | JOURNAL OF BACTERIOLOGY
 |
| 出版日期 | 2012-03-01 |
| 卷号 | 194期号:6页码:1342-1349 |
| 英文摘要 | ortho-Nitrophenol 2-monooxygenase (EC 1.14.13.31) from Alcaligenes sp. strain NyZ215 catalyzes monooxygenation of ortho-nitrophenol to form catechol via ortho-benzoquinone. Sequence analysis of this onpA-encoded enzyme revealed that it contained a flavin-binding monooxygenase domain and a heme-binding cytochrome b(5) domain. OnpA was purified to homogeneity as a His-tagged protein and was considered a monomer, as determined by gel filtration. FAD and heme were identified by high-performance liquid chromatography (HPLC) and HPLC-mass spectrometry (HPLC-MS) as cofactors in this enzyme, and quantitative analysis indicated that 1 mol of the purified recombinant OnpA contained 0.66 mol of FAD and 0.20 mol of heme. However, the enzyme activity of OnpA was increased by 60% and 450% after addition of FAD and hemin, respectively, suggesting that the optimal stoichiometry was 1:1:1. In addition, site-directed mutagenesis experiments confirmed that two highly conserved histidines located in the cytochrome b(5) domain were associated with binding of the heme, and the cytochrome b(5) domain was involved in the OnpA activity. These results indicate that OnpA is an unusual FAD-dependent monooxygenase containing a fused cytochrome b(5) domain that is essential for its activity. Therefore, we here demonstrate a link between cytochrome b(5) and flavin-dependent monooxygenases. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Microbiology |
| 研究领域[WOS] | Microbiology |
| 关键词[WOS] | NITRATE REDUCTASE ; SALICYLATE HYDROXYLASE ; ORTHO-NITROPHENOL ; SULFITE OXIDASE ; PURIFICATION ; HEME ; FLAVOPROTEIN ; SEQUENCE ; CLONING ; IDENTIFICATION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000300846900007 |
| 公开日期 | 2015-07-14 |
| 源URL | [http://ir.wipm.ac.cn/handle/112942/1615]  |
| 专题 | 武汉物理与数学研究所_磁共振应用研究部 |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, Key Lab Agr & Environm Microbiol, Wuhan, Peoples R China 2.Chinese Acad Sci, Wuhan Inst Phys & Math, Wuhan Ctr Magnet Resonance, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan, Peoples R China 3.Univ London Imperial Coll Sci Technol & Med, Dept Life Sci, London, England |
| 推荐引用方式 GB/T 7714 | Xiao, Yi,Liu, Ting-Ting,Dai, Hui,et al. OnpA, an Unusual Flavin-Dependent Monooxygenase Containing a Cytochrome b(5) Domain[J]. JOURNAL OF BACTERIOLOGY,2012,194(6):1342-1349. |
| APA | Xiao, Yi.,Liu, Ting-Ting.,Dai, Hui.,Zhang, Jun-Jie.,Liu, Hong.,...&Zhou, Ning-Yi.(2012).OnpA, an Unusual Flavin-Dependent Monooxygenase Containing a Cytochrome b(5) Domain.JOURNAL OF BACTERIOLOGY,194(6),1342-1349. |
| MLA | Xiao, Yi,et al."OnpA, an Unusual Flavin-Dependent Monooxygenase Containing a Cytochrome b(5) Domain".JOURNAL OF BACTERIOLOGY 194.6(2012):1342-1349. |
入库方式： OAI收割
来源：武汉物理与数学研究所
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