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Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N-2

文献类型：期刊论文

作者Wu, Meng-Ru2,6,7; Miao, Li-Li6,7; Liu, Ying6,7; Qian, Xin-Xin6,7; Hou, Ting-Ting2,6,7; Ai, Guo-Min6,7; Yu, Lu3; Ma, Lan2,6,7; Gao, Xi-Yan2,6,7; Qin, Ya-Ling2,6,7
刊名JOURNAL OF BIOLOGICAL CHEMISTRY
出版日期2022-09-01
卷号298
DOI10.1016/j.jbc.2022.102372
通讯作者Li, De-Feng(lidefeng@im.ac.cn) ; Liu, Zhi-Pei(liuzhp@im.ac.cn) ; Liu, Shuang-Jiang(liusj@sdu.edu.cn)
英文摘要Nitrogen (N-2) gas in the atmosphere is partially replenished by microbial denitrification of ammonia. Recent study has shown that Alcaligenes ammonioxydans oxidizes ammonia to dinitrogen via a process featuring the intermediate hydroxylamine, termed "Dirammox" (direct ammonia oxidation). However, the unique biochemistry of this process remains unknown. Here, we report an enzyme involved in Dirammox that catalyzes the conversion of hydroxylamine to N-2. We tested previously annotated proteins involved in redox reactions, DnfA, DnfB, and DnfC, to determine their ability to catalyze the oxidation of ammonia or hydroxylamine. Our results showed that none of these proteins bound to ammonia or catalyzed its oxidation; however, we did find DnfA bound to hydroxylamine. Further experiments demonstrated that, in the presence of NADH and FAD, DnfA catalyzed the conversion of N-15-labeled hydroxylamine to N-15(2). This conversion did not happen under oxygen (O-2)-free conditions. Thus, we concluded that DnfA encodes a hydroxylamine oxidase. We demonstrate that DnfA is not homologous to any known hydroxylamine oxidoreductases and contains a diiron center, which was shown to be involved in catalysis via electron paramagnetic resonance experiments. Furthermore, enzyme kinetics of DnfA were assayed, revealing a Km of 92.9 +/- 3.0 mu M for hydroxylamine and a kcat of 0.028 +/- 0.001 s(-1). Finally, we show that DnfA was localized in the cytoplasm and periplasm as well as in tubular membrane invaginations in HO-1 cells. To the best of our knowledge, we conclude that DnfA is the first enzyme discovered that catalyzes oxidation of hydroxylamine to N-2.
WOS关键词N-OXYGENASE AURF ; NITROSOMONAS-EUROPAEA ; NITROUS-OXIDE ; OXIDOREDUCTASE ; INTERMEDIATE ; NITRIFICATION
资助项目National Natural Science Foundation of China, China[91951101] ; National Natural Science Foundation of China, China[31861133002] ; National Natural Science Foundation of China, China[31870103] ; National Key R&D Program of China, China[2019YFA0905500]
WOS研究方向Biochemistry & Molecular Biology
语种英语
出版者ELSEVIER
WOS记录号WOS:000863323200009
资助机构National Natural Science Foundation of China, China ; National Key R&D Program of China, China
源URL[http://ir.hfcas.ac.cn:8080/handle/334002/129331]  
专题中国科学院合肥物质科学研究院
通讯作者Li, De-Feng; Liu, Zhi-Pei; Liu, Shuang-Jiang
作者单位1.Univ Sci & Technol China, Ctr BioAnalyt Chem, Affiliated Hosp 1, Hefei Natl Lab Phys Sci Microscale, Hefei, Peoples R China
2.Univ Chinese Acad Sci, Sch Life Sci, Beijing, Peoples R China
3.Chinese Acad Sci, High Magnet Field Lab, Hefei, Peoples R China
4.Shandong Univ, State Key Lab Microbial Technol, Qingdao, Peoples R China
5.Univ Sci & Technol China, Affiliated Hosp 1, Div Life Sci & Med, Hefei, Anhui, Peoples R China
6.Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing, Peoples R China
7.Chinese Acad Sci, Environm Microbiol Res Ctr, Inst Microbiol, Beijing, Peoples R China
推荐引用方式
GB/T 7714		 Wu, Meng-Ru,Miao, Li-Li,Liu, Ying,et al. Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N-2[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2022,298.
APA Wu, Meng-Ru.,Miao, Li-Li.,Liu, Ying.,Qian, Xin-Xin.,Hou, Ting-Ting.,...&Liu, Shuang-Jiang.(2022).Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N-2.JOURNAL OF BIOLOGICAL CHEMISTRY,298.
MLA Wu, Meng-Ru,et al."Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N-2".JOURNAL OF BIOLOGICAL CHEMISTRY 298(2022).

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来源：合肥物质科学研究院

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