FIGURE 5.

Complementation of the M. smegmatis ΔglgE single and ΔglgEΔtreS double mutants with GlgE variants. A and B, both genetic (A) and chemical (B) inactivation of TreS prevent M1P hyper-accumulation in the presence of exogenous trehalose. A, genetic inactivation of treS. The ΔglgEΔtreS mutant, which is insensitive to trehalose, was transformed with pVV16, pVV16_glgE, pVV16_glgE_Ala, or pVV16_glgE_Asp and grown with 10 mm trehalose. Plates were incubated at 37 °C for 4–5 days. B, chemical inactivation of TreS in the presence of validamycin A. The ΔglgE mutant was transformed with pVV16, pVV16_glgE, pVV16_glgE_Ala, or pVV16_glgE_Asp and grown in the presence of both validamycin A (10 mm) and trehalose (1 mm). Plates were incubated at 37 °C for 4–5 days.