IVD (Isovaleryl Coenzyme A dehydrogenase) is a mitochondrial matrix enzyme that belongs to the acyl-CoA dehydrogenase family. IVD is a homotetrameric flavoenzyme which catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA. Defects of the IVD gene lead to ineffective isoforms that are the underlying cause of isovaleric acidemia. Recombinant human IVD protein, fused to His-tag at N-terminus, was expressed in E.coli and purified by using conventional chromatography techniques.