Fig. 8From: Whole-genome and enzymatic analyses of an androstenedione-producing Mycobacterium strain with residual phytosterol-degrading pathwaysPreparation of soluble KstD211. a Group 4 contains one enzyme catalyzing dehydrogenation at the C1,2-position of 4-AD to generate ADD. b The KstD211 gene was amplified from the HGMS2 genome by PCR with a gradient annealing temperature. Arrows indicate expected PCR bands. c KstD211 was overexpressed in soluble form with a His6-tag at 18 °C. S, supernatant; P, pellets and M, molecular weight marker. d SDS-PAGE visualization of KstD211 purified by Ni–NTA affinity chromatography followed by MonoS chromatography. Lanes 1–8 represent samples from different fractionsBack to article page