FIG 7.
(A and B) GanB (A) and GanA (B) activities after 6 h of cultivation of B. subtilis strains, i.e., wt (KM0), ΔganR (BKE34170), ΔganS (KM468), ΔganA (KM589), and ΔganB (KM588), with different sugars in LB as described in Materials and Methods. GanB activity was measured in a 1:10 diluted culture medium toward the substrate AZCL galactan. GanA activity was determined in crude extracts toward pNP-β-Gal. (C) An electrophoretic mobility shift assay was carried out using the amplified 5′-end Cy5-Pgan DNA in the absence (control) and presence of GanR. The potential inducers of GanR were added to the reaction with the final concentration of 5 mM. (D) Thermal shift assay using purified GanR-Strep tagged in the absence of substrate (control) and in the presence of galactose, galactobiose, galactotriose, or galactotetraose with a final concentration of 1 mM.