title: Purification, crystallization and preliminary X-ray analysis of recombinant betaine aldehyde dehydrogenase 2 (OsBADH2), a protein involved in jasmine aroma, from Thai fragrant rice (Oryza sativa L.).
abstract: Fragrant rice (Oryza sativa L.) betaine aldehyde dehydrogenase 2 (OsBADH2) is a key enzyme in the synthesis of fragrance aroma compounds. The extremely low activity of OsBADH2 in catalyzing the oxidation of acetaldehyde is believed to be crucial for the accumulation of the volatile compound 2-acetyl-1-pyrroline (2AP) in many scented plants, including fragrant rice. Recombinant fragrant rice OsBADH2 was expressed in Escherichia coli as an N-terminal hexahistidine fusion protein, purified using Ni Sepharose affinity chromatography and crystallized using the microbatch method. Initial crystals were obtained within 24 h using 0.1 M Tris pH 8.5 with 30%(w/v) PEG 4000 and 0.2 M magnesium chloride as the precipitating agent at 291 K. Crystal quality was improved when the enzyme was cocrystallized with NAD(+). Improved crystals were grown in 0.1 M HEPES pH 7.4, 24%(w/v) PEG 4000 and 0.2 M ammonium chloride and diffracted to beyond 2.95 Å resolution after being cooled in a stream of N(2) immediately prior to X-ray diffraction experiments. The crystals belonged to space group C222(1), with unit-cell parameters a = 66.03, b = 183.94, c = 172.28 Å. An initial molecular-replacement solution has been obtained and refinement is in progress.
authors.affiliation: Synchrotron Light Research Institute (Public Organization), 111 University Avenue, Muang District, Nakhon Ratchasima 30000, Thailand. buabarn@slri.or.th
authors.forename: Buabarn
authors.initials: B
authors.lastname: Kuaprasert
authors.forename: Kun
authors.initials: K
authors.lastname: Silprasit
authors.forename: Natharinee
authors.initials: N
authors.lastname: Horata
authors.forename: Pongsak
authors.initials: P
authors.lastname: Khunrae
authors.forename: Ratree
authors.initials: R
authors.lastname: Wongpanya
authors.forename: Nonlawat
authors.initials: N
authors.lastname: Boonyalai
authors.forename: Apichart
authors.initials: A
authors.lastname: Vanavichit
authors.forename: Kiattawee
authors.initials: K
authors.lastname: Choowongkomon
mesh_terms: D050728:Betaine-Aldehyde Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D018360:Crystallography, X-Ray; D012275:Oryza / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D050728:Betaine-Aldehyde Dehydrogenase
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):22-5
references.pmid: 20057045
references.citation: Gene. 1991 Jul 15;103(1):45-52
references.pmid: 1879697
references.citation: Theor Appl Genet. 2011 Feb;122(3):533-41
references.pmid: 21046066
references.citation: FEBS Lett. 2009 Nov 19;583(22):3625-9
references.pmid: 19850038
references.citation: Acta Crystallogr D Biol Crystallogr. 2001 Oct;57(Pt 10):1445-50
references.pmid: 11567158
references.citation: Arch Biochem Biophys. 2010 Jan 1;493(1):71-81
references.pmid: 19766587
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):271-81
references.pmid: 21460445
references.citation: Bioinformatics. 2002 Sep;18(9):1250-6
references.pmid: 12217917
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
references.pmid: 21460454
references.citation: Biotechnol Adv. 2009 Jul-Aug;27(4):468-73
references.pmid: 19371779
references.citation: Acta Crystallogr D Biol Crystallogr. 1998 Jan 1;54(Pt 1):8-15
references.pmid: 9761813
references.citation: Plant Biotechnol J. 2005 May;3(3):363-70
references.pmid: 17129318
references.citation: J Mol Biol. 2004 Oct 8;343(1):29-41
references.pmid: 15381418
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Plant Mol Biol. 2008 Nov;68(4-5):439-49
references.pmid: 18704694
references.citation: J Mol Biol. 2010 Mar 05;396(4):870-82
references.pmid: 20026072