title: Expression, purification and preliminary X-ray crystallographic analysis of nitroalkane oxidase (NAO) from Pseudomonas aeruginosa.
abstract: Nitroalkane oxidase (NAO) is a flavin-dependent enzyme which catalyses the oxidation of nitroalkanes to the corresponding aldehydes or ketones, nitrite and hydrogen peroxide. In order to better understand the structure and function of this enzyme, NAO from Pseudomonas aeruginosa was purified and crystallized as a native and a selenomethionine-substituted (SeMet) enzyme. Both crystals diffracted to a resolution of 1.9 Å and belonged to the primitive orthorhombic space group P2₁, with unit-cell parameters a = 70.06, b = 55.43, c = 87.74 Å, β = 96.56° for native NAO and a = 69.89, b = 54.83, c = 88.20 Å, β = 95.79° for SeMet NAO. Assuming the presence of two molecules in the asymmetric unit in both crystals, the Matthews coefficients (VM) for native and SeMet NAO were calculated to be 2.30 and 2.48 Å³ Da⁻¹, with estimated solvent contents of 46.50 and 50.37%, respectively.
authors.affiliation: Division of Biotechnology, College of Life Sciences, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Jeong Hye
authors.initials: JH
authors.lastname: Lee
authors.forename: Ae Kyung
authors.initials: AK
authors.lastname: Park
authors.forename: Jae Soon
authors.initials: JS
authors.lastname: Oh
authors.forename: Ki Seog
authors.initials: KS
authors.lastname: Lee
authors.forename: Young Min
authors.initials: YM
authors.lastname: Chi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics*; D018360:Crystallography, X-Ray; D049308:Dioxygenases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D049308:Dioxygenases; C021876:2-nitropropane dioxygenase
keywords: NAO; Pseudomonas aeruginosa; nitro compounds; nitroalkane oxidase
references.citation: Appl Environ Microbiol. 1998 Mar;64(3):1029-33
references.pmid: 9501443
references.citation: J Bacteriol. 1978 Jan;133(1):53-8
references.pmid: 22538
references.citation: Bioorg Chem. 2004 Jun;32(3):125-39
references.pmid: 15110192
references.citation: Eur J Biochem. 2002 Dec;269(24):6302-7
references.pmid: 12473127
references.citation: Biochemistry. 2008 Dec 30;47(52):13745-53
references.pmid: 19053234
references.citation: Environ Mutagen. 1979;1(4):383-9
references.pmid: 399919
references.citation: Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
references.pmid: 12393927
references.citation: Biochem Biophys Res Commun. 2011 Feb 18;405(3):344-8
references.pmid: 21147069
references.citation: Ecotoxicol Environ Saf. 1992 Jun;23(3):253-9
references.pmid: 1376230
references.citation: Biochemistry. 2010 Jun 22;49(24):5035-41
references.pmid: 20481475
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: Biochemistry. 1998 Apr 28;37(17):6154-64
references.pmid: 9558355
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Biochemistry. 2006 Jan 31;45(4):1138-50
references.pmid: 16430210