title: Cloning, expression, purification, crystallization and X-ray crystallographic analysis of D-lactate dehydrogenase from Lactobacillus jensenii.
abstract: The thermostable D-lactate dehydrogenase from Lactobacillus jensenii (LjD-LDH) is a key enzyme for the production of the D-form of lactic acid from pyruvate concomitant with the oxidation of NADH to NAD(+). The polymers of lactic acid are used as biodegradable bioplastics. The LjD-LDH protein was crystallized using the hanging-drop vapour-diffusion method in the presence of 28%(w/v) polyethylene glycol 400, 100 mM Tris-HCl pH 9, 200 mM magnesium sulfate at 295 K. X-ray diffraction data were collected to a maximum resolution of 2.1 Å. The crystal belonged to space group P3121, with unit-cell parameters a = b = 90.5, c = 157.8 Å. With two molecules per asymmetric unit, the crystal volume per unit protein weight (VM) is 2.58 Å(3) Da(-1), which corresponds to a solvent content of approximately 52.3%. The structure was solved by single-wavelength anomalous dispersion using a selenomethionine derivative.
authors.affiliation: Structural and Molecular Biology Laboratory, School of Life Sciences and Biotechnology, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, Republic of Korea.
authors.forename: Sangwoo
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Kwangwoon University, Seoul 139-701, Republic of Korea.
authors.forename: Yong Hwan
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Structural and Molecular Biology Laboratory, School of Life Sciences and Biotechnology, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, Republic of Korea.
authors.forename: Kyung-Jin
authors.initials: KJ
authors.lastname: Kim
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007778:Lactobacillus / Q000201:enzymology*
chemical_list: D007770:L-Lactate Dehydrogenase
keywords: Lactobacillus jensenii; bioplastics; d-lactate dehydrogenase; polylactic acid
references.citation: Science. 2012 May 25;336(6084):1030-3
references.pmid: 22628654
references.citation: Int J Food Microbiol. 2003 Jun 15;83(2):115-31
references.pmid: 12706034
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: J Control Release. 2001 Apr 28;71(3):261-75
references.pmid: 11295219
references.citation: J Mol Biol. 2002 Apr 19;318(1):109-19
references.pmid: 12054772
references.citation: Acta Crystallogr D Biol Crystallogr. 2000 Aug;56(Pt 8):965-72
references.pmid: 10944333
references.citation: Acta Crystallogr D Biol Crystallogr. 2012 Apr;68(Pt 4):352-67
references.pmid: 22505256
references.citation: Eur J Biochem. 2000 Mar;267(6):1633-9
references.pmid: 10712593
references.citation: Acta Crystallogr D Biol Crystallogr. 1999 Apr;55(Pt 4):849-61
references.pmid: 10089316
references.citation: J Am Acad Dermatol. 2010 Mar;62(3):448-62
references.pmid: 20159311
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: J Mol Biol. 1997 Apr 4;267(3):640-60
references.pmid: 9126843
references.citation: J Mol Biol. 1994 Feb 25;236(3):759-85
references.pmid: 8114093