Fig. 5
Localization, purification and activity of recombinant M.tb PykA in different host strains. a–c Localization of rPykA-H in cellular fractions of E. coli BL21DE3 GroEL/ES strain, M. smegmatis and M.tb H37Ra respectively, detection was done using α-His antibodies. d Affinity purification of EC-PykA-H, analysis by 10 % SDS-PAGE e Western blotting with α-His antibody. f Affinity purification of MS-PykA-H, analysis by 10 % SDS-PAGE. g Western blotting with α-PykA and α-His antibodies. h Affinity purification of MT-PykA-H, analysis by 10 % SDS-PAGE. i Western blotting with α-His antibodies. Residual enzyme activity (in %) of EC-PykA-H, MS-PykA-H and MT-PykA-H in the j absence of cofactors or in the presence of k Mg2+ or l K+. Significance was determined by student’s unpaired t-test where *p < 0.05; **p < 0.01 and ***p < 0.001