Cloning, expression, purification, crystallization and X‐ray crystallographic analysis of D‐lactate dehydrogenase from Lactobacillus jensenii. Issue 8 (1st August 2014)

• Record Type: Journal Article
• Authors: Kim, Sangwoo; Kim, Yong Hwan; Kim, Kyung‐Jin
• Is Part Of: Acta crystallographica. Volume 70:Issue 8(2014:Aug.)
• Page Start: 1046
• Page End: 1048
• Publication Date: 2014-08-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X14012606
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The thermostable D‐lactate dehydrogenase from <italic>Lactobacillus jensenii</italic> (<italic>Lj</italic>D‐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 <italic>Lj</italic>D‐LDH protein was crystallized using the hanging‐drop vapour‐diffusion method in the presence of 28%(<italic>w</italic>/<italic>v</italic>) polyethylene glycol 400, 100 m<italic>M</italic> Tris–HCl pH 9, 200 m<italic>M</italic> 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 <italic>P</italic>3₁21, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 90.5, <italic>c</italic> = 157.8 Å. With two molecules per asymmetric unit, the crystal volume per unit protein weight (<italic>V</italic>_M) is 2.58 ų 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.</p> </abstract>