3MKH

Podospora anserina Nitroalkane Oxidase

3MKH の概要

• エントリーDOI: 10.2210/pdb3mkh/pdb
• 分子名称: NITROALKANE OXIDASE, FLAVIN-ADENINE DINUCLEOTIDE, MAGNESIUM ION, ... (5 entities in total)
• 機能のキーワード: oxidoreductase flavoenzyme, nitroalkane, acyl-coa dehydrogenase, fad, flavoprotein, oxidoreductase
• 由来する生物種: Podospora anserina
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 189277.12

構造登録者

Tormos, J.R.,Taylor, A.B.,Daubner, S.C.,Hart, P.J.,Fitzpatrick, P.F. (登録日: 2010-04-14, 公開日: 2010-06-02, 最終更新日: 2023-09-06)

主引用文献

Tormos, J.R.,Taylor, A.B.,Daubner, S.C.,Hart, P.J.,Fitzpatrick, P.F.
Identification of a hypothetical protein from Podospora anserina as a nitroalkane oxidase.
Biochemistry, 49:5035-5041, 2010

PubMed Abstract: The flavoprotein nitroalkane oxidase (NAO) from Fusarium oxysporum catalyzes the oxidation of primary and secondary nitroalkanes to their respective aldehydes and ketones. Structurally, the enzyme is a member of the acyl-CoA dehydrogenase superfamily. To date no enzymes other than that from F. oxysporum have been annotated as NAOs. To identify additional potential NAOs, the available database was searched for enzymes in which the active site residues Asp402, Arg409, and Ser276 were conserved. Of the several fungal enzymes identified in this fashion, PODANSg2158 from Podospora anserina was selected for expression and characterization. The recombinant enzyme is a flavoprotein with activity on nitroalkanes comparable to the F. oxysporum NAO, although the substrate specificity is somewhat different. Asp399, Arg406, and Ser273 in PODANSg2158 correspond to the active site triad in F. oxysporum NAO. The k(cat)/K(M)-pH profile with nitroethane shows a pK(a) of 5.9 that is assigned to Asp399 as the active site base. Mutation of Asp399 to asparagine decreases the k(cat)/K(M) value for nitroethane over 2 orders of magnitude. The R406K and S373A mutations decrease this kinetic parameter by 64- and 3-fold, respectively. The structure of PODANSg2158 has been determined at a resolution of 2.0 A, confirming its identification as an NAO.

PubMed: 20481475
DOI: 10.1021/bi100610e

実験手法

X-RAY DIFFRACTION (1.995 Å)