Cloning, expression, purification, crystallization and X-ray crystallographic analysis of (S)-3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum.
(S)-3-Hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD) is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA by the reduction of acetoacetyl-CoA to 3-hydroxybutyryl-CoA. The CbHBD protein was crystallized using the hanging-drop vapour-diffusion method in the presence of 2 M ammonium sulfate, 0.1 M CAPS pH 10.5, 0.2 M lithium sulfate at 295 K. X-ray diffraction data were collected to a maximum resolution of 2.3 Å on a synchrotron beamline. The crystal belonged to space group R3, with unit-cell parameters a = b = 148.5, c = 201.6 Å. With four molecules per asymmetric unit, the crystal volume per unit protein weight (VM) is 3.52 Å(3) Da(-1), which corresponds to a solvent content of approximately 65.04%. The structure was solved by the molecular-replacement method and refinement of the structure is in progress.
School of Life Sciences, KNU Creative BioResearch Group (BK21 Plus Program), Kyungpook National University, Daehak-ro 80, Daegu 702-701, Republic of Korea.
Eun-Jung
EJ
Kim
School of Life Sciences, KNU Creative BioResearch Group (BK21 Plus Program), Kyungpook National University, Daehak-ro 80, Daegu 702-701, Republic of Korea.
Kyung-Jin
KJ
Kim
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D003001:Cloning, Molecular; D046972:Clostridium butyricum / Q000201:enzymology*; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism
D011994:Recombinant Proteins; D015094:3-Hydroxyacyl CoA Dehydrogenases; C021388:3-hydroxybutyryl-CoA dehydrogenase
(S)-3-hydroxybutyryl-CoA dehydrogenase; Clostridium butyricum; n-butanol